Immunotoxicological effects of piperine in mice

doi:10.1016/j.tox.2003.10.006

Toxicology

Volume 196, Issue 3, 15 March 2004, Pages 229-236

https://doi.org/10.1016/j.tox.2003.10.006 Get rights and content

Abstract

The immunotoxicological effects of piperine were investigated in Swiss male mice, gavaged at a dose of 1.12, 2.25 or 4.5 mg/kg body weight for five consecutive days. All the dose levels had no overt toxic effect and the liver gained weight normally. Treatment at highest dose, however, resulted in significant decrease in the weight of spleen, thymus and mesenteric lymph nodes, but not of peripheral lymph nodes. All the dose levels suppressed the cellular population of lymphoid organs, except for the spleen, where the doses of 1.12 and 2.25 caused an increase. Haematologically, doses of 2.25 and 4.5 mg/kg caused a significant reduction in total leucocyte counts and differential leucocyte counts showed an increase in the percentage of neutrophils. The higher doses of 2.25 and 4.5 mg/kg suppressed the mitogenic response of B-lymphocyte to lipopolysaccharide. The number of primary antibody (IgM) forming cells in the spleen and the level of primary antibody in serum, was decreased. The doses of 1.12 and 2.25 mg/kg suppressed the mitogenic response of T-lymphocytes to phytohaemagglutinin and the nitroblue tetrazolium (NBT) dye reducing activity of peritoneal exudate cells (PECs). Since the lowest dose of 1.12 mg of piperine per kg body weight had no immunotoxic effect, it may be considered as immunologically safe “no observed adverse effect level (NOAEL)” dose.

Introduction

Piperine (1-piperoyl peperdine) is an alkaloid derived from black pepper (Piper nigrum Linn), commonly used worldwide as a spice due to the pungent and biting taste of piperine. Black pepper is also a folk lore medicine in many regions of the world, for the treatment of asthma, bronchitis, dysentry, pyrexia and insomnia (Rastogi and Mehrotra, 1991, Kirtekar and Basu, 1994). It is also claimed to exert varied pharmacological effects such as anti-fertility in rats and mice of both sexes (Munshi and Rao, 1972, Piyachaturawat and Chailurkit, 1992, Srivathana and Piyachaturawat, 1993), cardiovascular depression (Hatthakit et al., 1994), neurodepressant (Szolesanyi, 1983), anti-microbial and anti-parasitic (Raay et al., 1999), and modulation of carcinogen-induced oxidative stress (Khajuria et al., 1998). It has also been suggested to inhibit drug metabolising enzymes, resulting in non-specifically enhanced bio-availability of anti-tuberculosis drugs (Attal et al., 1985, Dalvi and Dalvi, 1991, Somashekhar, 2001).

The neurodepressant and anti-fertility effects of piperine, inhibition of cytochrome P₄₅₀ (Kirtekar and Basu, 1994), induction of chromosomal aberrations (John and Abraham, 1991) and presence in the spleen (Bhatt and Chandrashekhara, 1986), may result in adverse effects on the function of immunocompetent cells. The prolonged use of black pepper may result in toxic concentrations of piperine (Piyachaturawat et al., 1983). The purpose of the present study was to investigate the immunotoxic effects of piperine in mice.

Section snippets

Chemicals

Piperine, extracted from black pepper (P. nigrum Linn) of Indian origin, was obtained from Regional Research Laboratory , Jammu, India. All other chemicals used for experimental procedures were of analytic grade and purchased from HIMEDIA (India) and GIBCO (USA).

Animals and treatment

Six weeks old Swiss male mice (average weight 20±1 g), obtained from the animal breeding facility of Industrial Toxicology Research Centre, Lucknow, India, were randomly divided into four groups. The first group was gavaged with 0.2 ml of

Pathotoxicological studies

The treatment of animals with doses of 1.12 and 2.25 mg/kg, resulted in a significant increase in the weight of liver. The highest dose of 4.5 mg/kg, reversed this increase and the weight recorded was closer to control animals. However, all the doses had no significant effect on the weight of kidney and adrenals (Table 1). The treatment with highest dose of 4.5 mg/kg, resulted in a significant decease in the weight of spleen and thymus, but the lower doses of 1.12 and 2.25 mg/kg, caused an

Discussion

Immunological response of mice to piperine was studied by using both in vitro and in vivo parameters, such as changes in body weight and organs weight, haematology, histology and functional activity of immunocompetent cells (Munsen et al., 1982, Schuurman et al., 1994). In earlier animal studies, piperine given orally or intraperitoneally, was excreted slowly, and attained appreciable concentrations in various lymphoid organs and plasma (Bhatt and Chandrashekhara, 1986, Bajad et al., 2002). The

Acknowledgements

Authors are thankful to Director, Industrial Toxicology Research Centre, Lucknow, for his keen interest in the work, and to Dr. J. Singh, Scientist, Regional Research Laboratory, Jammu, India for providing the sample of piperine. Thanks are also due to Sri Lalji Shukla, Sri S.P. Dhruva for technical help and to Sri R.S. Verma for computer work.

References (38)

S.W Burchiel
The effect of environmental and other chemicals on the immune system: the emergence of immunotoxicology
Clin. Immunol.
(1999)
P Piyachaturawat et al.
Acute and sub ascute toxicity of piperine in mice, rats and hamsters
Toxicol. Lett.
(1983)
W Reanmongkol et al.
Effect of piperine on bioenergetic function of isolated rat liver mitochondria. Biochemical function of isolated rat liver mitochondria
Biochem. Pharmacol.
(1988)
J.G Vos et al.
Studies on 2,3,7,8-tetrachlorobenzo-p-dioxin induced immune suppression and decreased resistance to infection endotoxin hypersensitivity, serum zinc concentration and effect on thymosin treatment
Toxicology
(1978)
C.K Attal et al.
Biochemical basis of enhanced drug bioavailability by piperine. Evidence that piperine is a potent inhibitor of drug metabolism
J. Pharmacol. Exp. Therap.
(1985)
Bajad, S., Bedi, K.L., Singh, A.K., 1999. Effect of piperine a bioavailability enhances, on GIT: anti diarrhoeal...
S Bajad et al.
Simple high performance liquid chromatography method for the simultaneous detection of ketoconazole and piperine in rat plasma and hepatic culture
J. Chromatogr. A
(2002)
G.B Bhatt et al.
Studies on the metabolism of piperine: absorption, tissue distribution and excretion of urinary conjugates in rats
Toxicology
(1986)
D.I Birx et al.
The interference of T-cell activation by calcium channel blocking agents
J. Immunol.
(1984)
R.Y Cheung et al.
Permissive role of calcium in inhibition of T-cell mitogenesis by calmodulin antagonists
J. Immunol.
(1983)

A.G Cunningham et al.

Further improvements in the plaqueing technique for detecting single antibody forming cells

Immunology

(1968)

R.R Dalvi et al.

Differences in the effect of piperine and piperonyl butoxide on hepatic drug metabolisng enzyme system in rats

Drug Chem. Toxicol.

(1991)

Dean, J.H., Luster, Z.M.I., Munsen, A.E., Kimber, I., 1994. Immunotoxicology and Immunopharmacology, Second ed. Raven...

Descotes, J., 1988. Immunotoxicology of Drug and Chemicals. Elsevier,...

R.K.S Dogra et al.

Effect of dioctyl phthalate on immune system of rat

Indian J. Exp. Biol.

(1985)

R.K.S Dogra et al.

Immunomodulation due to coexposure to styrene and dioctyl phthalate in mice

Immunopharmacol. Immunotoxicol.

(1993)

U Hatthakit et al.

Cardiovascular effects of piperine in anaesthetised dog

Asia Pacific J. Pharmacol.

(1994)

Hudson, L., Hey, F.C., 1989. Practical Immunology, Third ed. Blackwell Scientific Publications,...

John, A.T., Abraham, S., 1991. Effect of black pepper on the incidence of chromosomal aberration in mouse bone marrow...

Cited by (60)

Alkaloids as Potential Multi-Target Drugs to Treat Alzheimer's Disease
2018, Studies in Natural Products Chemistry
Citation Excerpt :
The pharmacological properties of black or white pepper corns are attributed to piperine. Piperine (26) exhibits a wide spectrum of pharmacological activities including antimetastatic, antidepressant, hepatoprotective, and antitumor activities [167–170]. Piperine (26) has been found to be a potent inhibitor of both EeAChE (IC50 = 0.12 μM) and EqBChE (IC50 = 0.067 μM), displaying selectivity toward EqBChE [171,172].
Neurodegenerative diseases are characterized by the progressive and irreversible loss of neurons from specific regions of the brain. The pattern of neuronal loss is selective, affecting mainly the subcortical areas (nuclei of the base) and cerebral cortex, resulting in abnormality in the control of voluntary movement, impairment of memory, and cognitive ability. The progression of these debilitating and incurable conditions gives rise to dementia. The most known dementias are amyotrophic lateral sclerosis, Alzheimer's disease (AD), Parkinson's disease, and Huntington's disease. AD is the most prominent of these dementias, accounting for 60%–80% of the total dementia cases worldwide.
AD is a multifactorial, neurodegenerative, progressive, and fatal disorder characterized by loss of cholinergic neurons in the hippocampus and cerebral cortex mainly affecting cholinergic neurotransmission. The major pathological characteristics of AD include extracellular deposition of amyloid beta (Aβ) plaques, intracellular formation of neurofibrillary tangles (NFTs) comprising hyperphosphorylated and aggregated tau protein, neuroinflammation, and neurodegeneration. Based on the cholinergic hypothesis, that states that a low production of acetylcholine (ACh) initiates AD, the first drugs approved to treat AD were cholinesterase enzyme inhibitors (IChE), which are responsible for the hydrolysis of ACh. Despite being only palliative, so far the best treatment for AD is based on three FDA-approved cholinesterase inhibitors, namely, galanthamine rivastigmine, donepezil, and an N-methyl-d-aspartate receptor antagonist, memantine, indicated for joint therapy in the severe phase of AD.
New approaches including the observation of Aβ and NFTs lead to the amyloid and tau hypotheses as causes for AD development. Multi-target molecules able to inhibit cholinesterases and interfere with Aβ aggregation and/or tau protein hyperphosphorylation or neuroinflammation may be useful tools in the treatment of AD. Natural products have been a constant source of new approaches for the treatment of AD, especially plant alkaloids. For example, physostigmine (Physostigma venenosum), the prototype of rivastigmine, is an IChE and allosteric modulator of the central nicotinic receptor. Galanthamine (Galanthus woronowii) is a selective inhibitor of acetylcholinesterase, allosteric modulator of the central nicotinic receptor, inhibits Aβ aggregation, and promotes hippocampal neurogenesis. Berberis (Berberis mahonia) is an IChE, inhibits the formation and aggregation of Aβ, and reduces hyperphosphorylation of tau protein in some in vitro systems. Benzophenanthridine alkaloid chelerythrine (Chelidonium majus) is an IChE, inhibits Aβ aggregation, and induces Aβ disaggregation. The aim of this chapter is to review classes of alkaloid inhibitors of cholinesterases that have the potential to become multi-target prototypes for AD treatment.
An integrated strategy for rapid discovery and identification of the sequential piperine metabolites in rats using ultra high-performance liquid chromatography/high resolution mass spectrometery
2017, Journal of Pharmaceutical and Biomedical Analysis
Piperine, one of the major bioactive constituents isolated from natural flavorings and medicinal-culinary herbs, possesses various biological activities. In the present study, an integrated strategy based on ultra high-performance liquid chromatography/high resolution mass spectrometry was established to reveal piperine metabolism in rats. First of all, post-acquisition data-mining methods, including high resolution extracted ion chromatograms (HREICs) and multiple mass defect filtering (MMDF), were used to screen piperine metabolite candidates in a full-scan HRMS¹ level. Then, parent ion list-dynamic exclusion coupled with data-dependent data-acquisition method was utilized to acquire MSⁿ datasets. In addition, the established reverse molecular assembly (RMA) approach based on paired diagnostic product ions (pDPIs) coupled with neutral loss fragments (NLFs) was used to ascertain and identify the major-to-trace piperine metabolites efficiently. And then, the calculated ClogP values were utilized to distinguish the positional isomers. As a result, a total of 148 piperine metabolites were detected and characterized tentatively. The results demonstrated that piperine mainly underwent hydrogenation, dehydrogenation, hydroxylation, glucuronide conjugation, sulfate conjugation, ring-cleavage, and their composite reactions. Our results not only provided novel and useful data to better understand the safety, toxicity and efficacy of this potential therapeutic agent, but also indicated that the proposed strategy was reliable for a rapid discovery and identification drug-related constituents in vivo.
Neuroprotective Effects of Black Pepper and Its Bioactive Compounds in Age-Related Neurological Disorders
2023, Aging and Disease
Mechanism-Based Physiological Effects of Piperine: A Review
2023, Current Pharmacology Reports
Dietary supplementation of piperine improves innate immunity, growth performance, feed utilization and intestinal morphology of red seabream (Pagrus major)
2023, Fisheries and Aquatic Sciences
Piperine as a Potential Anti-cancer Agent: A Review on Preclinical Studies
2018, Current Medicinal Chemistry

View all citing articles on Scopus

View full text

Immunotoxicological effects of piperine in mice

Abstract

Introduction

Section snippets

Chemicals

Animals and treatment

Pathotoxicological studies

Discussion

Acknowledgements

Clin. Immunol.

Toxicol. Lett.

Biochem. Pharmacol.

Toxicology

Biochemical basis of enhanced drug bioavailability by piperine. Evidence that piperine is a potent inhibitor of drug metabolism

J. Pharmacol. Exp. Therap.

Simple high performance liquid chromatography method for the simultaneous detection of ketoconazole and piperine in rat plasma and hepatic culture

J. Chromatogr. A

Studies on the metabolism of piperine: absorption, tissue distribution and excretion of urinary conjugates in rats

Toxicology

The interference of T-cell activation by calcium channel blocking agents

J. Immunol.

Permissive role of calcium in inhibition of T-cell mitogenesis by calmodulin antagonists

J. Immunol.

Further improvements in the plaqueing technique for detecting single antibody forming cells

Immunology

Differences in the effect of piperine and piperonyl butoxide on hepatic drug metabolisng enzyme system in rats

Drug Chem. Toxicol.

Effect of dioctyl phthalate on immune system of rat

Indian J. Exp. Biol.

Immunomodulation due to coexposure to styrene and dioctyl phthalate in mice

Immunopharmacol. Immunotoxicol.

Cardiovascular effects of piperine in anaesthetised dog

Asia Pacific J. Pharmacol.