Abstract
Objective: This study aimed to evaluate histopathological and biochemical alterations caused by Piperine on the ovaries of adult female Wistar albino rats.
Methods: Thirty rats (3–4 months old, 200–250 g) were divided into three groups (n=10 each). Group A (control) received normal saline, Group B received Piperine at 5 mg/kg, and Group C received Piperine at 10 mg/kg body weight daily for 30 days. Serum estrogen and progesterone were measured using ELISA. Histology of ovarian tissues was done, and estrous cycle changes were monitored.
Results: Piperine exposure produced a dose-dependent increase in secondary follicle diameter (Group A: 210.4 ± 15.2 µm; Group B: 285.7 ± 18.9 µm; Group C: 342.1 ± 22.7 µm; p < 0.001). Serum estrogen was significantly elevated in Groups B and C (62.7 ± 5.3 pg/ml and 75.4 ± 6.1 pg/ml) compared to controls (48.2 ± 4.5 pg/ml), while progesterone declined (Group A: 1.45 ± 0.35 ng/ml; Group B: 1.01 ± 0.33 ng/ml; Group C: 0.41 ± 0.12 ng/ml; p < 0.001).
Conclusion: Piperine caused dose-dependent enlargement of secondary follicles with elevated estrogen and reduced progesterone, leading to arrest of the ovarian cycle at the proestrus stage. These findings suggest potential reproductive toxicity of piperine in females of reproductive age.
Keywords: Piperine, Secondary Follicles, Estrogen, Progesterone, Estrous cycle.
References
Umapathy VR, Dhanavel A, Kesavan R, Natarajan PM, S B, P V. Anticancer Potential of the Principal Constituent of Piper nigrum, Piperine: A Comprehensive Review. Cureus. 2024;16(2):1–8. https://doi.org/10.7759/cureus.54425
Tiwari A, Mahadik KR, Gabhe SY. Piperine: A comprehensive review of methods of isolation, purification, and biological properties. Med Drug Discov [Internet]. 2020;7:100027. https://doi.org/10.1016/j.medidd.2020.100027
Balakrishnan R, Azam S, Kim IS, Choi DK. Neuroprotective Effects of Black Pepper and Its Bioactive Compounds in Age-Related Neurological Disorders. Aging Dis 2023 Jun 1;14(3):750–77. https://doi.org/10.14336/AD.2022.1022
Adeyemo OA, Ore A, Ajisafe EO. The protective effect of piperine on oxidative stress and hepatic damage induced by diisononyl phthalate in rats. Egypt J Basic Appl Sci [Internet]. 2021 Jan 1;8(1):293–301. https://doi.org/10.1080/2314808X.2021.1983746
Wei M, LYu P, Li P, Hu J, Wu R, Ouyang Q, et al. Baolier Capsule’s Secret Weapon: Piperine Boosts Cholesterol Excretion to Combat Atherosclerosis. Drug Des Devel Ther [Internet]. 2024;18:6427–46. https://doi.org/10.2147/DDDT.S499598
Azam S, Park JY, Kim IS, Choi DK. Piperine and ItsMetabolite’s Pharmacology in Neurodegenerative and Neurological Diseases. Biomedicines. 2022;10(1):1–16. https://doi.org/10.3390/biomedicines10010154
Wu Z, Hu Y, Hao R, Li R, Lu X, Itale MW, et al. Research Progress of Genomics Applications in Secondary Metabolites of Medicinal Plants: A Case Study in Safflower. Int J Mol Sci [Internet]. 2025 Apr 19;26(8):3867. https://doi.org/10.3390/ijms26083867
Hasan R, Bhuia MS, Chowdhury R, Khan MA, Mazumder M, Yana NT, et al. Piperine exerts anti-inflammatory effects and antagonises the properties of celecoxib and ketoprofen: in vivo and molecular docking studies. Nat Prod Res 2024 Oct 11;1–16. https://doi.org/10.1080/14786419.2024.2413039
Srinivasan K. Black Pepper and its Pungent Principle-Piperine: A Review of Diverse Physiological Effects. Crit Rev Food Sci Nutr [Internet]. 2007 Oct 25;47(8):735–48. https://doi.org/10.1080/10408390601062054
Amir S, Shah STA, Mamoulakis C, Docea AO, Kalantzi OI, Zachariou A, et al. Endocrine Disruptors Acting on Estrogen and Androgen Pathways Cause Reproductive Disorders through Multiple Mechanisms: A Review. Int J Environ Res Public Health [. 2021 Feb 4;18(4):1464. https://doi.org/10.3390/ijerph18041464
Orozco-Galindo BV, Sánchez-Ramírez B, González-Trevizo CL, Castro-Valenzuela B, Varela-Rodríguez L, Burrola-Barraza ME. Folliculogenesis: A Cellular Crosstalk Mechanism. Curr Issues Mol Biol. 2025;47(2):1–16.https://doi.org/10.3390/cimb47020113
Marques P, De Sousa Lages A, Skorupskaite K, Rozario KS, Anderson RA, George JT. Physiology of GnRH and Gonadotrophin Secretion Endotext. 2000.
Land KL, Miller FG, Fugate AC, Hannon PR. The effects of endocrine-disrupting chemicals on ovarian- and ovulation-related fertility outcomes. Mol Reprod Dev. 2022;89(12):608–31. https://doi.org/10.1002/mrd.23652
Stojanović-Radić Z, Pejčić M, Dimitrijević M, Aleksić A, V. Anil Kumar N, Salehi B, et al. Piperine-A Major Principle of Black Pepper: A Review of Its Bioactivity and Studies. Appl Sci [Internet]. 2019 Oct 12;9(20):4270.https://doi.org/10.3390/app9204270
Charan J, Kantharia N. How to calculate sample size in animal studies? J Pharmacol Pharmacother. 2013;4(4):303–6. https://doi.org/10.4103/0976-500X.119726
Okafor IA, Nnamah US, Nnaka J. The fertility assessment of normal cyclic Wistar rats following the administration of methanolic extract of Portulaca oleracea: an experimental study. Middle East Fertil Soc J. 2021;26(1). https://doi.org/10.1186/s43043-020-00048-x
Sander A, Bival Štefan M, Sander T, Kučić Grgić D, Parlov Vuković J, Blažević I, et al. Characterization of Essential Oils and Ethanolic Extracts from Nine Pepper Species: Antioxidant and Antimicrobial Activity and Spectroscopic Analysis. Molecules. 2025;30(20):1–38. https://doi.org/10.3390/molecules30204140
Jahangir MA. Serum & Plasma Sampling from Small Animal Species Internationale Pharmaceutica Sciencia Serum & Plasma Sampling from Small Animal Species. 2025;(July):16–9. https://doi.org/10.31531/2231-5896.1000133
Al-Sabawy HB, Rahawi AM, Al-Mahmood SS. Standard techniques for formalin-fixed paraffin-embedded tissue: A Pathologist’s perspective. Iraqi J Vet Sci. 2021;35(1–3):935–43. https://doi.org/10.33899/IJVS.2021.131918.2023
20. Giovannopoulou E, Karakasi MV, Kouroupi M, Ieronimaki AI, Papakonstantinou E, Giatromanolaki A, et al. Ovarian Morphometric and Histologic Characteristics and Correlation with Clinical Factors: A Cross-Sectional Study. J Pers Med. 2023;13(2). https://doi.org/10.3390/jpm13020232
Siddiqui S, Khushtar M, Zafar A, Hasan SM, Arshad M, Ahmad MA, et al. Mechanism-Based Physiological Effects of Piperine: A Review. Curr Pharmacol Reports [Internet]. 2023 Mar 14;9(3):117–27. https://doi.org/10.1007/s40495-023-00314-2
Ziegenhagen R, Heimberg K, Lampen A, Hirsch-Ernst KI. Safety Aspects of the Use of Isolated Piperine Ingested as a Bolus. Foods [Internet]. 2021 Sep 8;10(9):2121. https://doi.org/10.3390/foods10092121
Ajayi AF, Akhigbe RE. Staging of the estrous cycle and induction of estrus in experimental rodents: an update. Fertil Res Pract. 2020;6(1):1–15. https://doi.org/10.1186/s40738-020-00074-3
Daware M, Mujumdar A, Ghaskadbi S. Reproductive Toxicity of Piperine in Swiss Albino Mice. Planta Med [Internet]. 2000 Dec 31;66(03):231–6.https://doi.org/ 10.1055/s-2000-8560
Yakubu J, Natsaridis E, du Toit T, Sousa Barata I, Tagit O, Pandey A. Curcumin and Piperine Nanoparticles inhibit CYP17A1 activity to Regulate Steroid Biosynthesis in Prostate Cancer. 2024. https://doi.org/10.20944/preprints202410.2072.v1
Singh KB. Persistent estrus rat models of polycystic ovary disease: an update. Fertil Steril 2005 Oct;84:1228–34. https://doi.org/10.1016/j.fertnstert.2005.06.013.

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