Pippali (Piper longum) Part 3

Pippali(Piper longum) Part 3

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General Pharmacology

Phytochemicals in Pippali (Piper longum) enable to enhance the bioavailability and absorption of certain drugs like Indomethacin, Vasicine, Diclofenac, Curcumin and several others. Piperine in Pippali (Piper longum) alters the membrane permeability and subsequently induces the synthesis of several membrane associated proteins involved in cytoskeletal functioning. [135]

Anti-inflammatory activity

The oil of dried fruit of Pippali (Piper longum) was well-known for its anti-inflammatory activity. To evaluate this,   carrageenan was injected to induce rat paw edema. The animals were divided in two groups. The first group was treated with Pippali (Piper longum) fruit oil. On oral administration at the dose of 0.5 ml/kg bodyweight, the essential oil of Pippali (Piper longum) fruit reduced the paw edema volume by 65.95% and at the dose of 1ml/kg bodyweight the oil reduced the paw edema volume by 72.34%. The second group was treated with ibuprofen at the dose of 100 mg/kg bodyweight. In this group of animals, ibuprofen reduced the paw edema volume by 70.21 %. This shows that at the dose of 1ml/kg bodyweight the oil of Pippali (Piper longum) fruit proved to be a better anti-inflammatory agent than ibuprofen. [136]    

For medicinal purposes, two varieties of Pippali (Piper longum) viz. Chhoti (small) Pippali and Badi (large) Pippali are commonly marketed. A study on rats showed that Chhoti (small) Pippali suppressed acute and sub-acute phases of inflammation while Badi (large) Pippali suppressed only acute phase of inflammation. Furthermore their anti-inflammatory activity was found to be superior to that of ibuprofen.

Carragenan induced edema shows two phases. The first phase attributed to release of histamine, 5-hydroxytryptamine and various kinins occurs in the first hour. The second phase related to the release of prostaglandin-like substances occurs in 2 to 3 hours. Both the varieties, Chhoti (small) Pippali and Badi (large) Pippali suppressed the paw edema by inhibiting these phlogistic mediators, and/or by stabilizing cell membrane.

Formaldehyde-induced inflammation occurs through proliferation and migration of fibroblasts which are mainly concerned with the formation of connective tissue. In the formaldehyde-induced edema model, both varieties suppress edema in 24 hours but the Chhoti variety was superior to Badi variety. The Chhoti variety at 200 mg/kg bodyweight produced more inhibition of edema than the standard anti-inflammatory drug, diclofenac sodium. [137]

By Ammonium sulphate precipitation method a protein was isolated from Pippali (Piper longum). This protein showed anti-inflammatory, antioxidant and free radical scavenging activity in vitro. At a dose of 1000μg/ml the Pippali (Piper longum) showed maximum anti-inflammatory activity which was similar to that of Diclofenac sodium. [138]

Antioxidant activity

Using aqueous extract of Pippali (Piper longum) fruit, silver nanoparticles were synthesized. These nanoparticles showed powerful antioxidant activities. Furthermore this activity was found to be similar to the standard antioxidants like vitamin E and butylated hydroxyanisole (BHA) [139]

Like silver-Pippali (Piper longum) nanoparticles, researchers have developed gold-Pippali (Piper longum) nanoparticles. The average size of the particle was 56 nano meter (nm). The shape of the particle was spherical and contained metallic gold. The particles show catalytic and antioxidant activities. [140]

In another study proteins were isolated from boiling water extract of Pippali (Piper longum). The antioxidant activity of proteins was analyzed using Hydroxyl radical scavenging assay and lipid peroxidation inhibition assay. The results were promising when compared with standard antioxidants Vitamin C, Vitamin E and butylated hydroxyanisole (BHA) [141]

Myocardial ischemia is a knotty medical problem. To evaluate efficacy of Pippali (Piper longum) in the treatment of this problem, myocardial ischemia was induced in rats by administration of isoproterenol. Petroleum ether extract of the root of Pippali (Piper longum) and piperine (one phytochemical found in Pippali) were administered. At 50% concentration the extract and piperine decreased lipid peroxidation and protected the myocardium from ischemic injury. This activity was attributed to the antioxidant property of Pippali (Piper longum). [142]  

In most studies water extracts of Pippali (Piper longum) were used to establish its antioxidant property. In one study both aqueous and methanolic extracts of seeds of Pippali (Piper longum) exhibited antioxidant activity. However methanolic extract of seeds of Pippali (Piper longum) demonstrated greater scale of antioxidant activity than the aqueous extract. [143]

Immunomodulatory activity

At a concentration of 500μg/ml the alcoholic extract of the fruits of Pippali (Piper longum) was found to have immunomodulatory property. In Balb/c mice, administration of the extract increased total white blood cell (WBC) count and bone marrow cellularity. [144]

Piperine the chief alkaloid isolated from Pippali (Piper longum) inhibits lipopolysaccharide (LPS)–induced tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β) and prostaglandin E-2 (PGE-2) production in murine microglial cell line BV-2. (BV-2 cells are used in laboratory to study neuro-inflammation). By acting as immunomodulator piperine modifies inflammatory response in nervous system, prevents neuro-dgeneration and is useful for the treatment of neuro-degenerative diseases. [145]  

Antiangiogenic activity of Pippali (Piper longum) extract was studied using B16F-10 melanoma cell. Intraperitoneal administration of 10mg/kg bodyweight of Pippali (Piper longum) extract to C57BL/6mice, inhibitied tumor directed capillaries by 50.6%. Moreover Pippali (Piper longum) inhibited vascular endothelial growth factor (VEGF)-induced tumor proliferation and cell migration. The extract had immunomodulatory effect on proinflammatory cytokines. The extract was non-toxic at concentrations of 10 μg/mL, 5 μg/mL and 1 μg/mL. [146]

Lipopolysaccharide (LPS) induces inflammatory response in bone-marrow-derived dendritic cells (BMDCs). Piperine inhibits this inflammatory response. Piperine inhibits expression of histocompatibility complex class II, CD40 and CD86 in bone-marrow-derived dendritic cells (BMDCs) in dose dependent manner. These findings provide insight into the immunological role of piperine. [147]

Antiallergic activity

Dahanukar et al (1984) and Chatterjee (1999) reported anti-allergic activity of Pippali (Piper longum). Amit et al reported the use of Pippali (Piper longum) in allergic rhinitis.

Mast cells release various mediators especially histamine associated with allergy. In an experimental study, albino rats were sensitized with horse serum. The rats were treated with ethanolic extract of Pippali (Piper longum) for 14 days. The result showed that the extract at 100 and 200 mg/kg bodyweight inhibited degranulation of mast cells to an extent of 62.44 and 67.24 % respectively. [148]

Researchers of Institute of Pharmaceutical Sciences, Kurukshetra University, Haryana, India, evaluated the anti-allergic activity of petroleum ether, alcoholic and aqueous extracts of fruit of Pippali (Piper longum). The extracts (100μg/mL) showed a significant antihistaminic activity. [149]

The milk extract of the fruits of Pippali (Piper longum), reduced passive cutaneous anaphylaxis in rats and protected guinea pigs against antigen-induced bronchospasm. [150] 

Antimicrobial activity     

    

The synthetic Pippali (Piper longum)-silver nanoparticles showed more potent antibacterial activity than the aqueous extract of the fruit of Pippali (Piper longum) [151]

A study describes the antibacterial activity of pure isolates from Pippali (Piper longum). Three isolates showed strong activity against Gram-positive bacteria and moderate activity against Gram-negative bacteria. Piperlongummine is active against Bacillus subtilis, Piperine against Staphylococcus aureus and Pellitorine against Bacillus sphaericus. [152]

The constituents isolated from the dry roots of Pippali (Piper longum) with n-hexane showed varying degree of antibacterial activity against many bacteria. The constituents showed better antibacterial profile than the n-hexane extract. The constituents showed antibacterial activity against Bacillus cereus and Escherichia coli. [153] 

The proteins isolated by 65% ammonium sulphate precipitate from Pippali (Piper longum) showed antibacterial activity against human pathogenic bacteria like Eschericia coli, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas, Salmonella typhimurium, various species of Streptococcus, Staphylococcus aureus and Vibrio cholerae. [154]

A study was undertaken to compare the in vitro antioxidant and antibacterial activity of chloroform, ethyl-acetate, hexane, ethanol and aqueous extracts of seeds of Pippali (Piper longum). The study showed that many bacteria were sensitive to chloroform, ethanol, ethyl-acetete and hexane extracts but not much to other extracts. [155]

From ethyl acetate extract of Pippali (Piper longum) plant the Indian researchers isolated and purified a well known alkaloid piperine. Piperine on further evaluation for antimicrobial activity was found to be effective against multi drug resistant Mycobacterium species. Interestingly piperine extracted from plant/fruit was more effective than other purified fractions isolated from Pippali (Piper longum) [156]

Antiviral activity

Respiratory infections most of the time are viral in origin. Chloroform and methonolic extracts of the seeds of Pippali (Piper longum) showed a very strong antiviral activity against para influenza virus. Of the two, the methanolic extract showed higher antiviral activity than that of chloroform extract. [157]

Hepatitis B is a naughty and knotty problem in the medical field. To one end it inflicts life-threatening insults like cirrhosis of the liver and hepatocellular carcinoma  on humans and to the other end; if detected at an early stage it can prove a trivial, regressible (if not totally ‘curable’ or ‘eradicable’) malady. Since the landmark research paper by Thyagarajan on eradication of hepatitis B virus by Bhoomyaamalakee (Phyllanthus amarus), there was a spate of research on herbs that can eradicate hepatitis B. Many herbs have now surfaced for the treatment of hepatitis B infection. Recently many pharmacologically active phytochemicals were isolated from various extracts of Pippali (Piper longum). Of these piperine possessed remarkable anti-hepatitis B virus activity. [158]   

Antifungal activity

When tested in vivo, the whole plant showed antifungal activity againgt Pyricularia oryzae, Rhizoctonia solani, Botrytis cineria, Phytophthora infestans, Puccinia recondite and Erysiphe graminis. A piperidine alkaloid, pipernonaline showed a potent fungicidal activity against Puccinia recondite while piperettine exhibited weak activity against Erysiphe graminis. They all are phytofungicidal agents. Whether phytopathogenic fungi are also pathogenic to humans is not clear. However Pippali (Piper longum) definitely shows antifungal activity against Candida albicans, a known human pathogenic fungus. [159], [160]

Antiparasitic activity

(1) Against Giardiasis

A group of 25 patients showing clinical signs and symptoms of giardiasis with stools positive for trophozoites and/or cysts of Giardia lamblia were treated with 1 g of Pippali (Piper longum) per day for 15 days. At the end of treatment there was a complete disappearance of trophozoites and/or cysts of Giardia lamblia from the stools of 23 patients. Mucus, pus cells and red blood cells were also absent from the stools. [161]

In another in vitro study on mice, the efficacy of aqueous extract of fruit powder of Pippali (Piper longum) and its ethanol extract were tested against experimental infection of Giardia lamblia. At the concentration of 125 μg/ml and 250 μg/mL the aqueous extract showed 100% antigiardial activity. Further fractionation in hexane and chloroform resulted in a total loss of activity. The antigiardial activity of Pippali (Piper longum) fruit powder in hexane, chloroform and n-butanol soluble fraction was comparable to standard antigiardial drugs.

(2) Against Amoebiasis

A study on rats suffering from caecal amoebiasis revealed that ethanolic extract, hexane fraction and n-butanol soluble fraction of the fruit of Pippali (Piper longum) at 1000 μg/mL and chloroform fraction at 500 μg/mL cured caecal amoebiasis. Furthermore the ethanolic plant extract and piperine cured 90% and 40% of rats with caecal amoebiasis respectively. Pippali (Piper longum) was also useful for the treatment of amoebiasis in vivo.

In a study on rats Ghoshal S and Lakshmi V showed that at 1000 μg/mL ethanolic extract of the roots of Pippali (Piper longum) cured 88% of caecal amoebiasis cases [163], [164] 

In an experimental study, caecal amoebiasis was induced in mice by injecting Entamoeba histolytica trophozoites directly into the caecum. The mice were then treated with oral administration of the Pippali (Piper longum) fruit extract, metronidazole (the standard antiamoebic drug) and a plain vehicle for consecutive five days. The results showed that at a dose of 1000 μg/kg bodyweight per day the Pippali (Piper longum) fruit extract showed 100% cure rate. At doses of 500 and 250 μg/kg bodyweight per day the extract was still effective and cured 93 and 46% of cases respectively. Metronidazole at doses of 125 and 62.5 mg/kg bodyweight cured 100 and 60% of cases respectively. [165]  

(3) Against Leishmania donovani

Piperlongumide and six other compounds found in Pippali (Piper longum) exhibit leishmanicidal activity against promastigotes (the extracellular forms in sandfly) and axenic amastigotes (the intracellular forms in vertebrates) of Leishmania donovani. [166]

Piperlongumine and some of its derivatives display leishmanicidal activity against promastigotes (the extracellular forms in sandfly) of Leishmania infantum and Leishmania amazonensis. This study suggests that piperlongumine and its derivatives may be antileishmanial drugs in future for the treatment of Leishmaniasis. [167]

(4) Anti-malarial activity

Many species of the genus Piper exhibit anti-Plasmodial (antimalarial) activity [168], [169]

(5) Activity against Filariasis

Using a series of organic solvents, from the pulverized fruits of Pippali (Piper longum), larvicidal components were isolated. Pipyahyine an isolated compound from the petroleum ether extract showed larvicidal activity against the filariasis vector Culex quinquefasciatus. Furthermore pipyahyine was found to be even more effective against filariasis than the parent extract of the plant. [170] 

Another study showed that pipernonaline an alkaloid in methanol extract of the fruit of Pippali (Piper longum) exhibited antilarval activity against mosquito larve of Culex pipiens pallens. The median lethal dose that killed larvae in 24 hours was 0.21 mg/litre. [171]

(6) Against Aedes aegypti

Ethanolic extract derived from Pippali (Piper longum) exhibited larvicidal activity against Aedes aegypti mosquitos. This mosquito is also known as yellow fever mosquito that can spread dengue fever, chickungunia, Zika fever and yellow fever.[172], [172] 

A crude methanol extract and hexane fraction derived from the fruits of Pippali (Piper longum) showed a strong larvicidal activity of 100% against Aedes aegypti mosquito larvae. This activity was attributed to pipernonaline. The larvicidal value LC (50) of pipernonaline was 0.25 mg/L. No larvicidal activity was observed with piperine, piperlongumine or piperettine. [173]

(7) Against Culex Mosquito

Various species of Culex mosquitoes transmit Arbovirus infections, Nematode infections, Filarial infection and Avian malaria. A larvicidal component isolated from the fruits of Pippali (Piper longum) kills larvae of Culex mosquitoes. This suggests that Pippali (Piper longum) can be considered as a powerful arsenal for the control of mosquito population. [174]  

(8) Anthelmintic activity

Strongyles, or alternatively, Strongyls are nematode worms of the family Strongylidae, order Strongylida. They are often parasitic in the gastrointestinal tract of mammals, especially grazers such as sheep, cattle and horse.

Amphistomes are flat worms commonly termed as flukes. They are trematodes. They are parasitic in sheep and humans.

Methanolic extract and its fractions from fruits of Pippali (Piper longum) were found to be highly active against strongyle ova, larvae and adult worms and amphistomes. [175]

For reasons not clear, in experimental studies in India; the anthelmintic activity of a drug is evaluated by testing them against Indian adult earth worms (Pheretima posthuma) and not against round worms (Ascaris lumbricoides). The anthelmintic property and potency of drugs is determined by time of onset of paralysis and time of death of worms.

To evaluate anthelmintic activity, the crude hydro-alcoholic extract of the fruit of Pippali (Piper longum) against earth worms (Pheretima posthuma), various concentrations (10, 25 and 50 mg/mL) of the fruit extract were tested in vitro. The extract induced spontaneous paralysis of earth worms (Pheretima posthuma). The anthelmintic activity of the fruit extract was superior to that of Albendazole. [176]  

In another anthelmintic study, the aqueous extract of 100 mg/mL concentration when administered to earth worms (Pheretima posthuma); induced paralysis in 2 minutes and worms died after 14 minutes. The results were compared with anthelmintic activity of Albendazloe. [177]

Anti-snake venom activities

Pippali (Piper longum) fruits have been traditionally used against snake bite in north-eastern and southern region of India. A study in fertilized chicken eggs, mice and rats showed that piperine from ethanolic extract of the fruit of Pippali (Piper longum) inhibited haemorrhagic action of Russell’s viper (Doboia russelii, Viperidae) in vitro and in vivo. The extract also inhibited defibrinogenating action, necrotizing action and lethal action of the venom. The extract inhibited venom induced paw oedema, degranulation of mast cells; and creatine kinase and catalase activity. [178]

Actions on skin

Melasma (dark, discolored patches on skin), freckles, and senile lentigines are hyperpigmentation disorders (hypermelanosis disorders) of the skin. Piperlonguminine a phytochemical in Pippali (Piper longum) is a potent melanogenesis inhibitor. Piperlonguminine inhibits α-melanocyte stimulating hormone (α-MSH)-induced melanogenesis. Piperlonguminine has no inhibitory effect on tyrosinase activity or a direct depigmenting effect of melanin. This activity of piperlonguminine is dose dependent. [179]

Exposure to ultraviolate rays causes trivial to severe skin disorders like erythema, oedema, skin burns, hyperpigmentation, photo-aging and photo-carcinogenesis. Piperine offers photoprotection to keratocytes in the skin. By inhibiting DNA damage and cell cycle arrest piperine prevents death of skin cells. These effects are said to be due to antioxidant and free radical scavenging property of piperine. [180]

Piperlonguminine inbibits the production of melanin in melanoma B 16 cell line. This effect was attributed to the inhibitory action of piperlonguminine on α-melanocyte stimulating hormone which in turn downregulates tyrosinase expression and melanin synthesis. [181]

An increase in the incidence of drug resistant melanoma is worrisome. Hence the need for novel effective therapeutic agents and treatment modalities. By elevation of the intracellular reactive oxygen species (ROS) formation, by inducing intracellular calcium homeostasis imbalance, DNA fragmentation and loss of mitochondrial membrane potential; piperine from Pippali (Piper longum) induces cell death in melanoma cells. Additionally piperine up-regulates the expression of apoptosis-inducing factor (AIF). Taken together, these results suggest that piperine could be a future phytochemical for the effective treatment of melanoma. [182]

Although piperine does not stimulate melanin synthesis, it promotes melanocyte proliferation. It is therefore used for the treatment of vitiligo. While on treatment with piperine a patient of vitiligo should avoid exposure to ultraviolet radiation (UVR) because the exposure causes photosentization. [183]

Exposure to ultraviolet radiation induces mutations in cutaneous cells. Additionally loss of activity in tumor suppressor gene, and overexpression of oncogenes in keratocytes result in the development of non-melanoma- skin cancers. Current topical therapies with 5-fluourouracil (5-FU), imiquimod, diclofenac, ingenol mebutate and photodynamic therapy are ineffective and inadequate as recurrence rate is very high. It is necessary that new therapies must target and clear clinically presenting and subclinical malignancies. Recent studies show that piperlongumine is effective in inducing cancer cell death without harming normal cells. [184] 

Actions on wound healing

Various species in the genus Piper have been reported to be beneficial for wound healing. However the specific reference with regard to Pippali (Piper longum) could not be cited.

Actions on Mouth

The fruits of Pippali (Piper longum) when taken orally have pungent taste, causes increased salivation and produces numbness of the mouth. [185]

Actions on the Breast

At 67 μg/ml/24hrs the synthetic Pippali (Piper longum)-silver nanoparticles showed a potent anticancer effect against MCF-7 breast cancer line. This anticancer activity was attributed to antioxidant property of the nanoparticles. [186] 

Actions on Hematopoetic system

The destruction of the cell membrane is the major factor for hemolysis. Various antioxidants, freeradical scavengers and stabilizers of cell membrane exert anti-hemolytic activity. A study showed that these properties of methanolic and aqueous extracts of Pippali (Piper longum) demonstrate a significant anti-hemolytic activity. [187]

A study showed that piperine, pipernonaline, piperlongumine and piperoctadecalidine prevent platelet aggregation induced by collagen, arachidonic acid and platelet-activating factor. Piperlongumine, in particular, showed the strongest antiplatelet aggregation activity. [188]

Ethanolic extract of Pippali (Piper longum) displayed protective effect on radiation induced damage on Swiss mice. The extract reduced the elevated levels of glutathione pyruvate transaminase (GTP), alkaline phosphatase (ALP) and lipid peroxidation (LPD) in liver and serum of radiation treated animals. [189]

A study showed that piperlongumine at concentrations of 10 and 20 μmol/L induced apoptosis in bone marrow mononuclear cells from patients with myeloid leukemias. By increasing intracellular reactive oxygen species (ROS) Pippali (Piper longum) induced apoptotic and autophagic death of primary myeloid leukemia cells. This anticancer activity of piperlongumine is dose and duration dependent. [190]

Actions on Musculoskeletal System

Administration of piperine from Pippali (Piper longum) to rats suffering from arthritis at doses of 20 and 100 mg/kg bodyweight per day for 8 days, relieved acute inflammation and joint pain. This study suggests that piperine can be a promising phytochemical for the treatment of rheumatoid arthritis (RA). [191]

Freund’s adjuvant is a solution of antigen emulsified in mineral oil. Freund’s Complete Adjuvant is composed of inactivated and dried mycobacteria, usually Mycobacterium tuberculosis whereas incomplete form lacks the mycobacterial components.

To evaluate anti-rheumatoid activity of Pippali (Piper longum), Complete Freund’s Adjuvant was used to induce arthritis in Wistar rats. Aqueous extract of the fruits of the plant was administered at doses of 200 and 400 mg/kg bodyweight. The administration of extract showed a significant reduction of in paw swelling on 4^th, 8^th, 14^th and 21^st day. These results were supported by radiographic analysis of affected knees. On 21^st day after induction of arthritis the dose of 400mg/kg bodyweight of Pippali (Piper longum) extract showed 46.32% improvement while Diclofenac sodium at the dose of 13.5 mg/kg bodyweight showed 55.00% improvement. These results suggest that Pippali (Piper longum) possesses useful activity for the treatment of arthritis. [192]    

In Ayurveda rheumatoid arthritis (RA) is known as ‘Aamawaata’. ‘Wardhamaan Pippali Rasaayana’ is a special and peculiar way of using Pippali (Piper longum) for the treatment of chronic ailments like ‘Aamawaata’, bronchial asthma, chronic liver diseases (fatty liver disease, cirrhosis of the liver etc.) In this, Pippali (Piper longum) is administered in gradually increasing doses up to certain days and then tapered in decreasing doses to stop the therapy. Soni A et al used ‘Wardhamaan Pippali Rasaayana’ to treat 73 patients of ‘Aamawaata’ i. e. rheumatoid arthritis (RA) with good results. [193]

Actions on Endocrine System

In one study the hydroalcoholic extract of Pippali (Piper longum) was administered to male mice at a dose of 200 mg/kg bodyweight for 30 days. The results showed that there was decrease in the function of pituitary-gonadal axis and decrease in spermatogenesis. [194]

Administration of piperine to adult male Swiss albino mice at the dose of 2.5 mg kg bodyweight orally for 15 days lowered thyroxin (T4) and triiodothyronine (T3). [195]    

Corticosterone can cause behavioral changes and depression. To study the beneficial effects of piperine against these ill effects, depression was induced in mice by injecting corticosterone for 3 weeks. The injection of corticosterone caused depression-like behavior in mice as indicated by the significant increase in immobility time and decrease in sucrose consumption. Moreover brain-derived neurotrophic factor (BDNF) and mRNA levels in hippocampus also decreased significantly. The treatment of these animals with piperine reversed these adverse changes. These effects are said to be mediated by increasing expression of brain-derived neurotrophic factor (BDNF) in the hippocampus. [196]  

Actions on Nervous System

Aqueous extract of Pippali (Piper longum) shows anti-stress activity. Pretreatment of adult Swiss albino mice at doses of 250mg/kg bodyweight and 500mg/kg bodyweight with Pippali (Piper longum) for 21 days ameliorated the stress-induced biochemical changes in the experimental animals.

In Sprague Dawley rats the aqueous extract of Pippali (Piper longum) demonstrated nootropic (cognition enhancing) activity.

Further the aqueous extract demonstrated anticonvulsant activity against strychnine, phenylenetetrazole (PTZ) and 4-amidopyridine induced convulsions. [197]

There are many causes of seizures. Epilepsy is the commonest one known to medical fraternity and lay persons. To evaluate the effects of anti-convulsant drugs, researchers use various methods to induce seizures. Vivek Sharma et al used ‘Audiogenic’ and ‘Maximal Electroshock’ stimuli to induce seizures. They found, aqueous and alcoholic extracts of fruits of Pippali (Piper longum) at 100 mg/kg bodyweight were useful to control theses seizures. The various mechanisms involved in ‘anti-seizure’ activity of Pippali (Piper longum) are:     

1. Antioxidant action:

Oxidative stress, free radical production can lead to initiation of lipid peroxidation, protein oxidation and DNA damage. These factors, via inactivation of glutamine synthesis lead to seizures. The antioxidant activity of Pippali (Piper longum) helps to control seizures.

2. Alteration in the levels of neurotransmitters

It is well known that decrease in gamma-amino-butyric acid (GABA) transmission has been implicated in excessive excitation that is characteristic of epilepsy. It is possible that extracts of Pippali (Piper longum) bind GABA sites in the brain, increase the density of GABA sites in the brain, reduce glutamate release and control seizures.  

3. Modulation of ion channels

Blockade of sodium-ion channels, inhibition of calcium channels and potentiation of GABA-induced chloride currents help control seizures. 

4. Activation of receptor potential

Transient receptor potential cation channel (TRPV1) is highly expressed in hippocampus, cerebral cortex, substantia nigra, hypothalamus and locus coeruleus. TRPV1 is involved in transmission and modulation of diverse stimuli. Administration of piperine at doses of 40 and 80 mg/kg bodyweight markedly delayed the onset of myoclonic jerks and generalized clonic seizures. 

 5. Inhibiting adenosinergic tone 

Extracts of Pippali (Piper longum) enhance endogenous adenosine levels in the CNS. Furthermore by reducing adenosine re-uptake, Pippali (Piper longum) extracts increase inhibitory adenosinergic tone to aid seizure suppression.  [198]

Recently an alkaloid piperine was isolated from the ethanolic extract of Pippali (Piper longum). Piperine is monoamine oxidase (MAO) inhibitor. This shows that piperine is a promising candidate for the treatment of mental depression. [199]

Oral administration of root powder of Pippali (Piper longum) to mice and rats at doses of 200, 400 and 800mg/kg bodyweight demonstrated a significant analgesic activity which was similar to NSAIDs (non steroidal anti-inflammatory agents). The dose of 800mg/kg bodyweight of Pippali (Piper longum) was found to be equivalent to the dose of 40 mg/kg bodyweight of Ibuprofen. The analgesic activity of Pippali (Piper longum) was much weaker than that of opioid analgesic pentazocine. [200] 

To evaluate the activity of Pippali (Piper longum) against cerebral ischemia in rats, the middle cerebral artery was occluded for 6 hours to induce cerebral ischemia. Pretreatment of the animals at doses of 100 and 200mg/kg bodyweight with dichloromethane fraction of Pippali (Piper longum) prevented the cerebral damage. This effect was attributed to anti-inflammatory, antioxidant and freeradical scavenging properties of the plant. [201] 

Administration of rotenone to rats, increases intracellular reactive oxygen species (ROS), induces motor deficit culminating into Parkinson’s disease (PD). Treatment of these rats with piperine and piperlongumine, alkaloids derived from Pippali (Piper longum), decreases reactive oxygen species (ROS), improves motor deficits and stabilizes mitochondrial membrane potential. Pretreatment of rats with alkaloids prevents the development of rotenone-induced Parkinsonism. [202]

Piperine exerts anxiolytic, antidepressant and antioxidant actions on Nervous system. Piperine is also a good memory enhancer. Taken together they prevent the formation of amyloid plaque in the brain. Thus Pippali (Piper longum) is useful for the prevention and treatment of Alzheimr’s disease. [203], [204]

Previously an alkaloid piperlongumine B had been isolated from Pippali (Piper longum). Recently piperlongumine B and its 19 analogs have been synthesized. Both the natural compound and synthetic analogs inhibit acetylcholinesterase. This research shows, these compounds may be promising drugs in future for the prevention and treatment of Alzheimer’s disease. [205]

The pain in peripheral nerve is due to direct stimulation of the sensory nerve fibers. In the late phase the pain is due to the inflammation and release of inflammatory mediators like histamine, prostaglandins (especially E2), bradykinins and serotonin. They evoke reversible calcium influx in sensory neurons sensitizing the sensory nerve to the sensation of pain. [206]    

At doses of 250 and 500 mg/kg bodyweight, methanol extract of Pippali (Piper longum) leaves exhibit peripheral analgesic activity in rats. [207]

Pain anywhere in the body triggers ‘stress’. At the dose of 250mg/kg bodyweight of methanolic extract of the fruit of Pippali (Piper longum) relieves ‘stress’. This anti-stress activity of Pippali (Piper longum) is attributed to anti-inflammatory and analgesic properties of piperine and piperlongumine found in Pippali (Piper longum). [208]

In an experimental study piperlongumine (PL) a natural alkaloid isolated from Pippali (Piper longum), by accumulating reactive oxygen species (ROS) in the cancer cells, selectively killed glioblastoma multiforme (GBM) cells but not normal cells. In cultures, piperlongumine (PL) could induce cell death in LN229, U87 and 8Mg glioblastoma multiforme (GBM) cell lines but not astrocytes.

On further exploration, piperlongumine (PL) was found to inhibit migration of LN 229 and U87 human glioblastoma cells but not normal astrocytes in the scratch-wound cultural model. [209], [210]

For evaluation of anti-glioma activity of Pippali (Piper longum), the experimental glioma model was developed in rats using C6 glioma cells. The glioma-induced animals showed increased level of lipid peroxides (LPO), lactate dehydrogenase (LDH), creatine kinase (CK), 5’nucleotidase (5’ ND) and acetylcholine esterase (AChE). Treatment of glioma induced rats with 20 mg/kg bodyweight of Pippali (Piper longum) significantly altered these biochemical changes suggesting anticancer effect of Pippali (Piper longum). The anticancer effect of Pippali (Piper longum) was confirmed by microscopic analysis. Inerestingly there was no toxic effect on normal brain cells. [211]

Actions on Respiratory System

Along with hypnotic effect, morphine and pentobarbitone exert respiratory depressant effect. In frogs, mice, rats and dogs, piperine showed central stimulant effect and antagonized respiratory depression induced by morphine and pentobarbitone.

Nalorphine is known to antagonize respiratory depression induced by morphine. The anti-respiratory-depressant effect of petroleum ether extract of the fruit of Pippali (Piper longum) was comparable to that nalorphine. However unlike nalorphine, piperine did not antagonize morphine-induced analgesia in rats.  

In smaller doses the petroleum ether extract of the fruit of Pippali (Piper longum) produced respiratory stimulant effect but in larger doses it caused convulsions in laboratory animals. This may be due to some medullary stimulant factors in the extract. [212], [213], 214]

Pippali (Piper longum) stabilizes mast cells and prevents the release of histamine and other pro-inflammatory chemicals. The effect of petroleum ether, alcoholic extracts and decoction of the fruits of Pippali (Piper longum) was studied for antihistaminic activity on Guinea pigs. At the dose of 100 μ g/kg bodyweight the extracts significantly inhibited the release of histamine from mast cells. The extracts at 50, 100 and 200 mg/kg bodyweight protected the animals from histamine induced bronchospasm. This effect was dose dependent. Thus Pippali (Piper longum) prevents the development of bronchial asthma. In milk-induced leukocytosis, petroleum ether extract and decoction at the dose of 200mg/kg bodyweight decreased the number of leukocytes significantly but the alcoholic extract did not show the desired effect. [215]

Actions on Cardiovascular System

Adriamycin an important anticancer chemotherapeutic drug is cardiotoxic. To determine cardioprotective activity of Pippali (Piper longum) against adriamycin (ADR) cardiotoxicity; by administering 15 mg/kg bodyweight of adriamycin (ADR) to Wistar rats cardiotoxicity was induced. Methanolic extract of Pippali (Piper longum) was administered at doses of 250 mg/kg and 500 mg/kg bodyweight for 21 days. The result showed that the extract significantly reduced the adriamycin-induced toxicity. This effect was attributed to antioxidant property of Pippali (Piper longum). [216]

In an experimental study myocardial infarction was induced by subcutaneous injection of isoproterenol in Wistar albino rats. Methanolic extract of Pippali (Piper longum) was then administered for 28 days. The result showed decrease in the levels of enzymes and improvement in myocardium. Pretreatment with Pippali (Piper longum) extract prevented the myocardial infarction. [217]

To study phytopharmacology of Pippali (Piper longum) the research was funded by National Research Council of Thailand. In the study, ethanolic extract of Pippali (Piper longum) was administered to rats by intravenous route at 1, 5 and 10 mg/kg bodyweight. Researchers found a significant reduction in systolic and diastolic blood pressure, mean arterial pressure and heart rate. Although this effect was observed at all doses, the effect was dependent on dose. The extract at 0.005 to 0.5mg/kg bodyweight induced relaxation in endothelium in intact and denuded aorta. The results indicated that ethanolic extract of Pippali (Piper longum) has vasodilator and hypotensive effect. This involved both nitric oxide and prostacyclin pathways as well as inhibition of entrance of extracellular calcium into the arterial smooth muscles. [218]

In an experimental study, treatment of rats at the dose of 40mg/kg bodyweight in drinking water for 4 weeks; with NG-Nitro-L-arginine methyl ester (L-NAME) caused sustained decrease in the nitrite/nitrate (NOx) concentration in plasma, vasoconstriction and hypertension. Treatment with piperine from Pippali (Piper longum) restored the concentration of NO metabolites. Moreover piperine restored the levels of superoxide dismutase, catalase and glutathione peroxidase and decreased the levels of peroxidation markers. The treatment also brought back the elevated blood pressure to normal. These results were attributed to the antioxidant activity of piperine. [219]

Endothelial dysfunction exists in diabetics. Methanolic extract of Pippali (Piper longum) fruit (MEPL) was administered at doses of 22.5, 45 and 90 mg/kg bodyweight to diabetic rats for 28 days. The result displayed a significant decrease in relaxant effect of acetylcholine (Ach) on isolated aorta from diabetic rats. This suggests methanolic extract of Pippali (Piper longum) fruit (MEPL) is useful to treat endothelial dysfunction and hypertension in diabetics. However more study is necessary in this regard. [220]

Dehydropipernonaline, an amide isolated from the fruit of Pippali (Piper longum) has demonstrated the ability to induce coronary vasodilatation. [221] 

Actions on GI Tract

The crude extract of Pippali (Piper longum) as well as piplartine, suppresses the ciliary movements of the oesophagus of the frog, which may be due to the suppression of cough reflex. [222]

At the dose of 4.5 mg/kg bodyweight, piplartine reduced the basal gastric acid secretion, as well as that stimulated by pentagastrin in rabbits. Thus piplartine prevents acid peptic disease and development of peptic ulcer in rabbits. [223]

Trikatu (three pungent or acrid substances having medicinal properties) is a peculiar Ayurvedic formulation. It contains equal quantities of Shunthee-dried ginger-(Zingiber officinale), Marichee-black pepper-(Piper nigram) and Pippali-long pepper-(Piper longum). Giant fresh water prawns (Macrobrachium rosenbergii), the experimental animals in this study, were fed with trikatu at a concentration of 5% in each feed for a period of consecutive 60 days. The results showed a significant improvement in survival and growth performance. The animals showed weight gain, increase in growth rate, elevation in activities of digestive enzymes; increased levels of proteins, vitamins C and E and minerals sodium, potassium. This study showed that by enhancing the secretion of digestive enzymes, trikatu acted as appetizer which facilitated efficient digestion, absorption of nutrients which improved general health and growth. There was a better build up of muscle mass. This suggests that trikatu has its own, special influence on protein synthesis i.e. non-steroidal, non-hormonal anabolic activity. [224]

The alkaloid piperlongumine isolated from Pippali (Piper longum) selectively kills gastric cancer cells while sparing normal cells. This is due to anti-inflammatory and antioxidant properties of piperlongumine. [225]

Infection of gastric mucosa by Helicobacter pylori (H. pylori) causes hyperacidity and gastric ulceration that culminate into gastric carcinoma. Administration of piperine impairs Interleukin-8 (IL-8) secretion in Helicobacter pylori (H. pylori) infeted gastric cells, suppresses the entry of Helicobacter pylori (H. pylori) toxin into gastric epithelium, decreases the motility of the bacteria in Helicobacter pylori (H. pylori) infeted gastric cells and decreases the adhesion of Helicobacter pylori (H. pylori) to gastric epithelium. Thus piperine blocks hyperacidity and subsequent development of gastric ulceration and the risk of oncogenesis. [226]

Mongolian gerbil or Mongolian jird is a small rodent. It is most commonly kept as a small house pet in England and America or as an experimental animal. To evaluate anti-Helicobacter pylori (anti-H. pylori) activity of piperine, the male Mongolian gerbils were infected with Helicobacter pylori (H. pylori). Treatment of the infected animals with piperine eradicated the infection from the pyloric antrum and cured Helicobacter pylori (H. pylori) induced gastritis. Furthermore pretreatment of animals with piperine prevented the development of gastritis and oncogenesis. [227]  

Expression of genes flagE and flagA is a trigger for the development of gastric carcinoma. By suppressing adhesion of Helicobacter pylori to gastric epithelium and expression of flagE and flagA genes piperine not only prevents the development of gastric adenocarcinoma but also arrests its further growth. [228]

In an experimental study 5 % acetic acid was used intra rectally to induce ulcerative colitis/inflammatory bowel disease (IBD) in mice. The disease activity was estimated by weight loss, stool consistency, gross or occult bleeding, length of colon affected and histological changes. Administration of piperine from Pippali (Piper longum) significantly reversed the disease process and histological changes. Furthermore piperine inhibited secretion of pro-inflammatory mediators. The study also showed that preadministration of piperine prevented the development of ulcerative colitis/inflammatory bowel disease (IBD) [229]

Piperlongumine inhibits the growth of colon cancer cells in time and dose dependent manner. This suggests that in future piperlongumine may be a promising drug for the treatment of colon cancer. [230]

A study showed that piperine, an alkaloid found in the fruits of black pepper (Piper nigrum) and Pippali (Piper longum) inhibited the growth of HRT-18, human adenocarcinoma by causing the apoptosis in the cancer cells. This effect was dependent on the dose of the drug. [231]

Actions on the Liver

(1) Hepatoprotective activity

Carbon tetrachloride (C Cl4) is a known hepatotoxic agent that causes liver fibrosis. A study showed that administration of ethanolic extract of Pippali (Piper longum Linn) reduced hepatic fibrosis. The liver functions also returned to normal as was evident by the normalization of the levels of various liver enzymes. [232]  

In another study ethanol, petroleum ether, solvent ether, ethyl acetate, butanol and butanone extracts of the fruits of pippali (Piper longum) were evaluated for their hepatoprotective activities in adult Wistar rats. The ethanolic and butanol fractions showed a significant hepatoprotective activity. The results were compared to control and Liv-52-treated rats. [233]

Following treatment of Wistar rats with milk extract of Pippali (Piper longum) fruit and root powder orally at dose of 200 mg/day for 21 days Jagruti A. Patel et al observed a significant hepatoprotective effect in carbon tetrachloride (CCl4)-induced hepatic damage. This effect was comparable to silymarin 25mg/kg bodyweight per day for 21 days. [234]

(2) Actions on viral hepatitis

In modern medicine, "infective jaundice" is known as viral hepatitis. Jaundice is caused by Hepatitis A and Hepatitis E viruses. Both are ribonucleic acid (RNA) viruses. Although viral hepatitis is said to be a self-limiting condition, in some patients it can pose lifethreatening problems. Modern medicine has no satisfactory treatment for ‘jaundice’ (viral hepatitis). Anti-inflammatory, antioxidant, free radical scavenging, immunomodulatory and hepatoprotective activities of Pippali (Piper longum) can alter the course and reduce the duration of the disease. [235]

 

[For more information on Ayurvedic treatment of ‘jaundice’ i. e. viral hepatitis, please refer to pharmacology of Kutakee (Picrorrhiza kurroa) by the same author.]

 

For anti-hepatitis B activity of Pippali (Piper longum) please read above, antiviral activity of Pippali (Piper longum)

 

(3) Actions on Fatty liver/ Cirrhosis of the liver   

 

The endoplasmic reticulum is a network of sac-like structures held together by cytoskeleton. It plays an important role in the production, processing and transport of proteins and lipids. To cope up with the stress, cells activate an intracellular signaling pathway-the unfolded protein response (UPR). The unfolded protein response (UPR) has three aims: (1) restoration of normal function of cells (2) degrading misfolded proteins and (3) activating the signaling pathways. If these objectives are not achieved within a certain time span or if the disruption is prolonged the unfolded protein response (UPR) aims towards apoptosis. [236]    

 

The unfolded protein response (UPR) is activated in several liver diseases like viral hepatitis, alcohol-induced liver injury and non alcoholic fatty liver disease (NAFLD) all of which are associated with steatosis and may be linked to unresolved endoplasmic stress. If this is true, restoration of endoplasmic stress homeostasis prior to endoplasmic stress-induced cell death may provide a therapeutic rationale in these diseases. It is plaussible that piperine, piperlongumine and Pippali (Piper longum) restore endoplasmic stress homeostasis to control or cure liver diseases.     

 

‘Wardhamaan Pippali Rasaayana’ is a special, peculiar and unique way of using Pippali (Piper longum) for the treatment of chronic ailments like ‘Aamawaata’ (Rheumatoid Arthritis), bronchial asthma, chronic liver diseases (fatty liver disease, cirrhosis of the liver etc.) In this regimen, Pippali (Piper longum) is administered in gradually increasing doses till it reaches upto maximum therapeutic dose and then tapered in decreasing doses till the dose reduces to nil.

 

The treatment schedule: Every day in the morning a quarter glass of milk is given on empty stomach. Ten minutes later Pippali (Piper longum) fruit powder mixed with 100-150 mL of milk is administered orally as a single dose followed by half a glass of milk. The diet is restricted to low protein and moderate quantity of carbohydrates, a thin liquid food: rice or oats boiled in milk or water (gruel or porridge). If the patient feels thirsty diluted milk is allowed to quench the thirst. Approximately two litres of milk are administered every day. In other words the patient is on ‘milk diet’. This schedule supplies balanced nutrition and restricts fluid intake to required level.

         

Usually many patients tolerate this regimen. Some patients may complain of burning sensation in the stomach, flatulence, milk diarrhea (lactose intolerance) and sleeplessness. These symptoms disappear as the treatment progresses.

 

In chronic liver diseases protein synthesis by liver is disturbed. Usually they manifest protein deficiency with low albumin. Milk diet supplies ample amount of lactalbumin. This also helps reduce intrahepatic, extrahepatic and intra-abdominal fat deposition.

 

The usual practice is to administer 3 grams of Pippali (Piper longum) fruit powder on the first day, increase the dose by 3 grams per day from the second day till the dose reaches 30 grams on the tenth day; then from the eleventh day taper the dose by 3 grams per day till it becomes zero grams. The dose pattern is shown in the table:

 

 

The table showing dose pattern of ‘Wardhamaana Pippali Rasaayana’

 

Day	No. of grams		Day	No. of grams
1	3		11	27
2	6		12	24
3	9		13	21
4	12		14	18
5	15		15	15
6	18		16	12
7	21		17	9
8	24		18	6
9	27		19	3
10	30		20	0

  [237] 

(4) Hepatic cancers

By increasing intracellular levels of reactive oxygen species (ROS) piperlongumine (PL) selectively kills hepatocellular carcinomas (HCC) but not normal hepatocytes. Piperlongumine (PL) also inhibits the invasion and migration of cancer cells. [238]

Actions on pancreas

Piperine inhibits lipopolysaccharide-induced inflammatory response in pancreas. Piperine also reduces the severity of cerulean-induced acute pancreatitis (AP). Piperine reduces myeloperoxidase activity, reduces the elevated serum levels of amylase, lipase and trypsin. Piperine also reduces the histologic damage that happens in pancreatitis. [239]

Piperlongumine isolated from the pippali (Piper longum) fruits was used alone or in combination with gemcitabine in vitro and in xenograft mouse model to treat pancreatic cancers. Piperlongumine inhibited the proliferation of pancreatic cancer cell lines and potentiated the apoptotic effects of gemcitabide. [240]

Actions on Metabolism

In rats fed with high fat diet, as expected, total serum cholesterol was elevated. Methyl piperine (Methyl piperate) inhibited the elevation of serum cholesterol.

The unsaponifiable fraction of the oil of Pippali (Piper longum) also decreased the total cholesterol and hepatic cholesterol in hypercholesterolaemic mice. [241], [242]

The antihyperlipidemic action of the fruit of Pippali (Piper longum) was attributed to piperine, piperlongumine and pipernonaline isolated from the ethanolic extract of the plant. Their antihyperlipidemic action was comparable to commercially available antihyperlipidemic drug simvastatin. [243]

A study was undertaken to explore the effect of piperine from Pippali (Piper longum) in obesity-induced dyslipidemia. Male Sprague Dawley rats were fed on high fat diet for eight weeks to induce obesity and obesity-induced dyslipidemia. Later they were treated with piperine 40 mg/kg bodyweight and sibutramine 5 mg/kg bodyweight for three weeks with the continuation of high fat diet. Results showed that inspite of high fat diet, piperine reduced bodyweight, body fat mass, triglyceride, total cholesterol, LDL cholesterol and VLDL cholesterol. Piperine also increased the level of HDL. Melanocortin-4 is a protein that reduces appetite and hence the food intake. The structure of piperine resembles that of melanocortin-4. Researchers feel piperine is melanocortin-4 agonist. So by reducing appetite and food intake piperine exerts its antiobesity action. [244]

Actions against diabetes

Due to antioxidant property, Pippali (Piper longum) shows potent hypoglycemic activity in alloxan induced diabetic rats. Pippali (Piper longum) can also be used to treat complications of diabetes associated with oxidative stress. [245]

Oil of Pippali (Piper longum) at 100 and 200 mg/kg bodyweight and piperine at 25 and 50 mg/kg bodyweight administered for 28 days to streptozotocin-induced diabetic rats, reduced blood glucose levels. There was significant increase in bodyweight, liver glycogen content, plasma insulin and high-density lipoprotein and decrease in glycosylated haemoglobin, triglyceride and total plasma cholesterol. [246]

Actions on Urinary System

At concentrations of 25, 50 and 100 μg/ 100 μL, piperlongumine showed antibacterial activity against various bacteria causing urinary tract infection. [247]

Different extracts of the fruit of Pippali (Piper longum) were evaluated for their inhibitory effect on formation of calcium oxalate stones in the urinary tract. The result showed aqueous and alcoholic extracts had higher capacity to inhibit the crystal formation and aggregation as compared to ethyl acetate and petroleum ether extracts. Researchers think this effect might be   due to the high concentration of alkaloids in aqueous and alcoholic extracts. [248]

Liu D et al demonstrated that by elevating reactive oxygen species (ROS), in vitro piperlongumine from Pippali (Piper longum) suppressed the proliferation, invasion and migration of bladder cancer cells and in vivo suppressed the development and growth of bladder cancer cells. The epithelial mesenchymal transition is required for nuclear reprogramming. This transition plays an important role in cancer progression. This is a new mechanism of oncogenesis. Piperlongumine inhibited epithelial mesenchymal transition. This suggests a novel mechanism underlying anticancer effect of Pippali (Piper longum). This research can provide a new strategy for bladder cancer therapy. [249]

Actions on Male Reproductive System

In male albino rats, by decreasing testicular hormone synthesis piperine significantly impaired spermatogenesis. The histological study supported the suppression of spermatogenesis. These effects were temporary and reversible. After stopping piperine treatment the spermatogenesis returned to normal. This study suggests that piperine can be used as male contraceptive agent. [250]

In another animal experiment, piperine was administered to mature male albino rats at doses of 5 and 10 mg/kg body weight for 30 days. The dose of 10 mg/kg body weight of piperine treatment caused a significant reduction in the weight of testis and accessory sex organs. Histological studies revealed that piperine at a dose of 5 mg/kg bodyweight caused a partial degeneration of germ cells, whereas piperine at the dose of 10mg/kg bodyweight caused severe damage to the seminiferous tubules, decrease in Leyding cell nuclear diameter and desquamation spermatocytes and spermatids. Correlated to the structural changes, there was a fall in epididymal sperm concentration. The dose of 10 mg/kg bodyweight of piperine also caused a marked increase in serum gonadotropins and a decrease in intratesticular testosterone concentration. [251]

To evaluate the effect of piperine, an alkaloid present in Pippali (Piper longum), on the testis and reproductive functions, piperine was administered to adult male rats at 1, 10, 100 mg/kg bodyweight for 3o days. A significant decrease in the activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase (i. e. antioxidant enzymes). A dose dependent increase in lipid peroxidation and hydrogen peroxide generation was also observed. These observations show that piperine induces oxidative stress in the testis that triggers apoptosis in the testis culminating into impaired reproductive function. [252]  

To study the effect of piperine, an alkaloid present in Pippali (Piper longum) on the epididymis, piperine was administered to adult male rats at 1, 10, 100 mg/kg bodyweight for 3o days. The results showed that at doses of 10 and 100 mg/kg bodyweight sperm count and motility of sperms decreased while at the dose of 100 mg/kg bodyweight the viability of sperms decreased significantly. Piperine also decreased the activities of antioxidant enzymes in the epididymis. [253]

Piperine was found to inhibit proliferation of human prostate cancer DU145, PC-3 and LNCaP cells. Piperine treatment also induced autophagy in PC-3 and LNCaP cells. [254]

Actions on Female Reproductive System 

To study effects of Pippali (Piper longum) on female reproductive system, hexane fraction of the plant was administered to adult female rats at doses of 150 and 250mg/kg bodyweight for 30 days. The result showed that at the dose of 250 mg/kg bodyweight, the length of estrous cycle prolonged and there was reduction in the number of implantation sites. Histopathology of the uterus showed degeneration of uterine glands and endometrial epithelial cells. The Graafian follicle in the ovary showed loss of cumulus oocyte complex. The serum levels of leutinizing hormone (LH) and follicle stimulating hormone (FSH) increased. The ovarian cytokines, nitric oxide and COX-2 levels decreased two fold. The levels of antioxidant enzymes also reduced. This suggests that via gonadotrophin insuffiency and modulation of inflammatory mediators, Pippali (Piper longum) induces infertility in female rats. It is not clear whether these changes are temporary and reversible. If reversible, then Pippali (Piper longum) can be used as a female contraceptive agent. [255]

V Lakshmi et al showed that crude extract of the powder of the fruit of Pippali (Piper longum) exhibited 100 % antifertility effect in female rats 1 to 7 days post coitum. They also showed that while hexane fraction showed 86 % efficacy, 1-butanol soluble, 1-butanol insoluble and chloroform fractions were inactive. [256]

An infusion of the root of Pippali (Piper longum) has been used to expel retained placenta after child birth. [257]

Piperlongumine (PL) a natural alkaloid from Pippali (Piper longum) possesses a highly selective anticancer activity. By enhancing accumulation of intracellular reactive oxygen species (ROS) it induces apoptosis in cancer cells in G2/M phase in dose and time dependent manner. Piperlongumine (PL) shows anticancer activity against ovarian cancers. Piperlongumine in combination with cisplatin or paclitaxel has synergistic antigrowth effect on human ovarian cancer cells. [258]

Antitumor activity

Piperlongumine also known as piplartine has shown a potent anticancer activity against many types of cancers. Piperlongumine has been found to be proapototic, anti-invasive and antiangiogenic agent. Piperlongumine synergizes with many anticancer chemotherapeutic agents. Researchers have now successfully synthesized many analogues of piperlongumine and piperlongumine-based hybrid anticancer compounds. [259]

At concentration of 500μg/ml the alcoholic extract of the fruits of Pippali (Piper longum) was toxic to Dalton’s lymphoma ascites (DLA) cells and at 250 μg/ml to Ehrlich ascites carcinoma (EAC) cells [260]

Recently researchers investigated extensively the anticancer activity of the fruit of Pippali (Piper longum) against human cancer cell lines (DU145 prostate, A549 lung, THP-1 leukemia, IGR-OVI-1 ovary and MCF-7 breast). The study was carried out in vitro and in vivo by using chloroform, acetone, benzene, hexane, ethyl alcohol and aqueous extracts. As flavonoides are important anticancer agents their concentrations in these extracts were determined. While hexane extract exhibited 90 to 92 % cytotoxicity against most of the cell lines tested; benzene, hexane and acetone extracts demonstrated 91 to 95 % cytotoxicity against A549 lung cancer. [261]

N. B. Nair unraveled the secret behind anti-cancer property of Pippali (Piper longum). He identified piperlongumine (PL) as anti-cancer phytochemical in the plant. Furthermore he showed that piperlongumine (PL) was effective against primary brain tumors, breast, lung, colon and prostate cancers; leukemia and lymphoma.

The cytotoxicity of piperlongumine has been attributed to increase in concentration of reactive oxygen species (ROS) in cancer cells. [262], [263]

Toxicity and Safety Profile

Pippali (Piper longum) is safe to use for long term therapeutic usage. A study showed that a single dose of 3G/kg bodyweight administered to Charles foster rats for 90 days revealed no adverse effects. Studies in mice about LD₅₀ values of piperine, piperlongumine and piperlonguminine were reported as 56.2 +/- 3.0, 110.1 +/- 7.8 and 115.3 +/- 9.5 mg/kg bodyweight respectively. Administration of a single dose of 3 to 5G/kg body weight did not show any acute toxicity, mortality or morbidity in experimental animals. However Pippali (Piper longum) and its isolated chemicals should not be used in pregnancy and lactation because of potential interactions. [264]

Formulations and Preparations

Ayurvedic Formulations containing Pippali (Piper longum)

Pippali (Piper longum), having pleotropic pharmacological activities is much sought-after herb. No wonder it is one of the ingredients in more than 300 Ayurvedic medicinal formulations. It is beyond scope of this work to elaborate all of them. Here I elaborate some important and commonly used formulations.   

No	Ayurvedic Name of the Yoga  (Formulation)	Ayurvedic Indications and Uses	Dose	References
1	Aamalakee-Pippali Rasaayana Amalaki-Pippali Rasayan)	Adaptogen, Anti-aging, Antioxidant Anti-fatigue, Build up stamina, Digestant, Immunomodulator	1-3 Grams BD with warm water or cow’s ghee	C. Chi. 1, 2/7
2	Pippalyaadi Ghrita	Jeerna Jwara (Chronic fever), Kshaya (emaciation, wasting, cachexia), Kaasa (cough), Paarshwa Shoola (backachake)	¼ to ½ teaspoon BD before food	C. Chi. 3, 3/219, 17/36-38, ayurinfo. com
3	Sitopalaadi Choorna	Kaasa (cough),bronchitis Shwasa (bronchial asthma), Tuberculosis Immunomodulator	Infants: 100-250 mg BD with honey, Children: 250 mg to 1 Gram BD with honey, Adults: 1 to 3 Grams BD with honey	C. Chi. 8/103 www.ayurtimes.com
4	Panchkola Choorna/ Panchkolaasaw/ Panchkola Ghrita	Deepana (appetizer), Paachana (digestive), Swarabhanga(hoarseness of voice), Colics, Gulma (tumour),	3 Grams OD or BD with buttermilk	Yogaratnakar Paribhaasha 1-2, https://ayurmedinfo.com
5	Pippalyaadi Yavagu (medicated gruel)	Yoni Vyapat (various vaginal disorders), Shoola (colics), Hridroga (heart  disorders)	Depends on disease	C. Chi. 30/54
6	Tilwaka Ghrita	Udara (ascites), Vidradhi (abscess), Gulma (tumour), Unmaada (hysteria)	Depends on appetite	S. Chi. 14/7
7	Jeewantyaadi Choorna	Kaasa (cough, bronchitis), Shwaasa (bronchial asthma), Hikkaa (hiccup), Jwara (PUO), Parshwashoola (backache, colics)	½ to 1 tea spoon with warm water or honey after food	A. H. Chi. 3/160, Sahasrayoga Choorna-Prakarana, www.ayurpages.com
8	Soorana Wataka, Soorana Gutika	Arsha (piles, fissures in ano), Apachana (indigestion), Kaasa (cough, bronchitis), Shwaasa (bronchial asthma), Hikkaa (hiccup), rasaayana (rejuvenative)	1 to 2 tablets 1 to 2 times a day before food	A. H. Chi. 5/33 https://ayurmedinfo.com
9	Tikta Ghrita	Panchkarma (external use) and as medicine for internal use. For Wrana- chikitsa (wound healing), Twachaa Roga (skin disorders), Visarpa (herpes zoster), Shwitra (leucoderma), Netra Roga (ophthalmic disorders), Hridroga (heart diseases), Arsha (piles, fissures in ano)	¼ to ½ teaspoon with warm water, before or after food, twice a day	A. H. Chi. 19/2-7, https://ayurmedinfo.com
10	Dhanwantari Ghrita	Shoth (oedema), Madhumeha (diabetes mellitus), Waata-Rakta (gout), Twachaa-Roga (skin disorders), Mano-Roga (psychiatric disorders)	¼ to ½ teaspoon with warm water, before or after food, twice a day	A. H. .Chikitsa Sthana, 8/157;  12/19-23, https://ayurmedinfo.com
11	Daadimaadi Ghrita	Paandu (anaemia), Hridroga (heart diseases), Atisaar (diarrhea), Stree-Wandhyatwa (female infertility), Snehana procedure. Can be safely given up to 6 to 8 weeks	¼ to ½ teaspoon with warm water, before or after food, twice a day	A. H. Chi. 13/2-4, 15/40, C. Chi. 16/44-46 https://www.planetayurveda.com, https://ayurmedinfo.com
12	Pippali Rasaayana	Kaasa (cough, bronchitis) Shwaasa (bronchial asthma), Tuberculosis Immunomodulator	For details see above	A. H. U. 39/96 https://ayurmedinfo.com
13	Jaatiphalaadi Choorna	Atisaar (diarrhea), Grahanee (dysentery, colitis),Udar shooal (colics), Kaasa (cough, bronchitis) Shwaasa (bronchial asthma), Tuberculosis Immunomodulator	1 to 3 Grams once or twice a day before food	B. P. S. 4/48-51, https://ayurmedinfo. com
14	Samashakara Choorna	Agnimaandya (loss of appetite), Apachana (indigestion), Pratishyaaya (common cold)  Kaasa (cough, bronchitis) Shwaasa (bronchial asthma), Kanthashosha (dry throat), Walaya/Kshawathu/Adhrusha (sore-throat), Arsha Roga (piles, fissures in ano)	3 Grams OD or BD Can be taken safely up to 2 to 4 months	B. R. (Arshroga  Adhikaar) 306-310 , https://ayurmedinfo.com
15	Ashtakatwara Tailam	Urustambha (aorto-iliac occlusion), Grudhrasee/Gridhrasee (sciatica)	20 ml orally  BD before lunch and dinner, also for local application	ejbps, 2016, Volume 3, Issue 10, 502-506
16	Pippalyaasawa	Pandu (anaemia), Agnimaandya (loss of appetite), Apachana (indigestion), Yakrut-Pleeha Roga (hepatic disorders, spleenic disorders, portal hypertension), Kaasa (cough, bronchitis) Shwaasa (bronchial asthma), Kshaya (tuberculosis), Gulma (tumours), Grahanee (colitis, irritable bowel disease), Arsha (piles, fissures in ano)	Safe for children above 2 years, Dose: 1 to 2 ml  For adults, Dose: 15 to 25 ml Can be safely used upto 3 to 4 months	S. S. M. 10/28-33, B. R. AFI Volume 1, https://ayurinfo.com, https://  www.planetayurveda.com/library
17	Pippalyaadi Tailam	Arsha (piles, fissures in ano),  Manyaastambha (cervical spondylosis) Panchakarma chikitsa (massage), enema	For external use	B. R. (Arshoroga Adhikaara) 115-118 https://ayurmedinfo.com
18	Pippalyaadi Varti (The wick soaked in Pippali-Ghritam or Tailam)	Yoni-daaha(vaginitis) Yoni-Vyapat (vaginal disease)	The medicated wick to be inserted in the vagina	www.iamj. in
19	Pippali Khanda/ Pippali-khanda Awaleha	Amlapitta (acid peptic disorder, GERD), Ahita aahaar sewana (improper-irregular dietary habits), Agnimaandya (loss of appetite), Apachana (indigestion)	3 to 10  Grams BD, 15 minutes before food	B. R. 53, 121- 125, iajm.co.in/ vol.8, No.3; July-September 2017, https:// www.asiapharmaceutics. info
20	Trikatu Choorna	Amlapitta (acid peptic disorder, GERD), Ahita aahaar sewana (improper-irregular dietary habits), Agnimaandya (loss of appetite), Apachana (indigestion), Pandu (anaemia), Agnimaandya (loss of appetite), Apachana (indigestion), Yakrut-Pleeha Roga (hepatic disorders, spleenic disorders, portal hypertension), Kaasa (cough, bronchitis) Shwaasa (bronchial asthma), Kshaya (tuberculosis), Gulma (tumours), Grahanee (colitis, irritable bowel disease), Arsha (piles, fissures in ano) , Medowriddhi (obesity, dyslipidemia), Madhumeha (diabetes)	1 to 3 Grams, BD, before food, as lambative with honey	https:// www.ayurtimes.com, https://easyayurveda.com, www.planetayurveda.com/library, www.ayurpages.com/trikatu-choorna

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