Volume : 13, Issue : 09, September – 2026
Title:
PHARMACOLOGICAL EVALUATION OF STANDARDIZED EXTRACT OF BLACK PEPPER ON SODIUM ARSENITE-INDUCED REPRODUCTIVE TOXICITY IN EXPERIMENTAL RATS
Authors :
Ms. Monika Popat Barhate*, Dr. Kamble H. V., Prof. Ghodake S. R., Dr. Nikam A. R.
Abstract :
Background: Sodium arsenite is an environmental toxicant capable of producing oxidative stress and impairment of male reproductive function. The present study investigated the protective effect of standardized Piper nigrum (black pepper) extract against sodium arsenite-induced reproductive toxicity in experimental rats.
Methods: Animals were randomly divided into six groups (n = 6): normal control, sodium arsenite control, CoQ10 (10 mg/kg), and standardized P. nigrum extract-treated groups receiving 50, 100, or 200 mg/kg. Sodium arsenite was administered orally at 10 mg/kg. Body weight and reproductive parameters were assessed, including absolute and relative weights of the testes, epididymis, and prostate, sperm count, sperm motility, dead and abnormal sperm, and serum testosterone. Testicular homogenates were evaluated for total protein, superoxide dismutase (SOD), reduced glutathione (GSH), malondialdehyde (MDA), and nitric oxide (NO). Testicular histopathology was also examined.
Results: Sodium arsenite produced significant reproductive and biochemical alterations, including reductions in body and reproductive-organ weights, sperm count, sperm motility, testosterone, SOD, GSH, and testicular protein, together with increases in dead and abnormal sperm, MDA, and NO. The 200 mg/kg dose of standardized black pepper extract significantly attenuated most of these changes compared with the sodium arsenite control group. Histopathological examination further demonstrated reduced inflammatory infiltration, necrosis, Leydig cell damage, and spermatogenic-cell vacuolation following treatment.
Conclusion: Standardized P. nigrum extract, particularly at 200 mg/kg, demonstrated protective activity against sodium arsenite-induced reproductive toxicity, potentially through attenuation of oxidative and nitrosative stress and preservation of testicular structure and reproductive function. Further mechanistic and safety studies are warranted.
Keywords: Piper nigrum; black pepper; sodium arsenite; reproductive toxicity; oxidative stress; sperm quality; testosterone; testis; SOD; GSH; MDA; nitric oxide.
Cite This Article:
Please cite this article in press Monika Popat Barhate et al., Pharmacological Evaluation Of Standardized Extract Of Black Pepper On Sodium Arsenite-Induced Reproductive Toxicity In Experimental Rats.,. Indo Am. J. P. Sci, 2026; 13(09).,
REFERENCES:
1. Kim YJ, Kim JM. Arsenic toxicity in male reproduction and development. Dev Reprod. 2015;19(4):167-180. doi:10.12717/DR.2015.19.4.167.
2. Abdul KS, Jayasinghe SS, Chandana EPS, Jayasumana C, de Silva PMCS. Arsenic and human health effects: a review. Environ Toxicol Pharmacol. 2015;40(3):828-846. doi:10.1016/j.etap.2015.09.016.
3. Kumar S, Malhotra S, Prasad AK, Van der Eycken EV, Bracke ME, Stetler-Stevenson WG, et al. Anti-inflammatory and antioxidant properties of Piper species: a perspective from screening to molecular mechanisms. Curr Top Med Chem. 2015;15(9):886-893. doi:10.2174/1568026615666150220120651.
4. Parim BN, Harishankar N, Balaji M, Pothana S, Sajjalaguddam RR. Effects of Piper nigrum extracts: restorative perspectives of high-fat diet-induced changes on lipid profile, body composition, and hormones in Sprague-Dawley rats. Pharm Biol. 2015;53(9):1318-1328. doi:10.3109/13880209.2014.980585.
5. Souza ACF, Marchesi SC, Ferraz RP, Lima GDA, Oliveira JA, Machado-Neves M. Effects of sodium arsenate and arsenite on male reproductive functions in Wistar rats. J Toxicol Environ Health A. 2016;79(6):274-286. doi:10.1080/15287394.2016.1150926.
6. Huang Q, Luo L, Alamdar A, Zhang J, Liu L, Tian M, et al. Integrated proteomics and metabolomics analysis of rat testis: mechanism of arsenic-induced male reproductive toxicity. Sci Rep. 2016;6:32518. doi:10.1038/srep32518.
7. Ince S, Avdatek F, Demirel HH, Arslan-Acaroz D, Goksel E, Kucukkurt I. Ameliorative effect of polydatin on oxidative stress-mediated testicular damage by chronic arsenic exposure in rats. Andrologia. 2016;48(5):518-524. doi:10.1111/and.12472.
8. Chinta G, Periyasamy L. Reversible anti-spermatogenic effect of piperine on epididymis and seminal vesicles of albino rats. Drug Res (Stuttg). 2016;66(8):420-426. doi:10.1055/s-0042-108186.
9. Adedara IA, Abolaji AO, Awogbindin IO, Farombi EO. Suppression of the brain-pituitary-testicular axis function following acute arsenic and manganese co-exposure and withdrawal in rats. J Trace Elem Med Biol. 2017;39:21-29. doi:10.1016/j.jtemb.2016.07.001.
10. Zubair M, Ahmad M, Qureshi ZI. Review on arsenic-induced toxicity in male reproductive system and its amelioration. Andrologia. 2017;49(9):e12791. doi:10.1111/and.12791.
11. Ramos ATA, et al. Morphological and morphometrical changes on adult Wistar rat testis caused by chronic sodium arsenite exposure. Environ Sci Pollut Res Int. 2017;24(36):27905-27912. doi:10.1007/s11356-017-0200-2.
12. Wagner H, Cheng JW, Ko EY. Role of reactive oxygen species in male infertility: an updated review of literature. Arab J Urol. 2017;16(1):35-43. doi:10.1016/j.aju.2017.11.001.
13. Majzoub A, Agarwal A. Systematic review of antioxidant types and doses in male infertility: benefits on semen parameters, advanced sperm function, assisted reproduction and live-birth rate. Arab J Urol. 2018;16(1):113-124. doi:10.1016/j.aju.2017.11.013.
14. Prathima P, Pavani R, Sukeerthi S, Sainath SB. α-Lipoic acid inhibits testicular and epididymal oxidative damage and improves fertility efficacy in arsenic-intoxicated rats. J Biochem Mol Toxicol. 2018;32(2):e22016. doi:10.1002/jbt.22016.
15. Lima GDA, Sertorio MN, Souza ACF, Menezes TP, Mouro VGS, Gonçalves NM, et al. Fertility in male rats: disentangling adverse effects of arsenic compounds. Reprod Toxicol. 2018;78:130-140. doi:10.1016/j.reprotox.2018.04.015.
16. Chen X, Ge F, Liu J, Bao S, Chen Y, Li Y, et al. Diverged effects of piperine on testicular development: stimulating Leydig cell development but inhibiting spermatogenesis in rats. Front Pharmacol. 2018;9:244. doi:10.3389/fphar.2018.00244.
17. Bastaki M, Aubanel M, Bauter M, Cachet T, Demyttenaere J, Diop MM, et al. Absence of adverse effects following administration of piperine in the diet of Sprague-Dawley rats for 90 days. Food Chem Toxicol. 2018;120:213-221. doi:10.1016/j.fct.2018.06.055.
18. Guvvala PR, Ravindra JP, Selvaraju S, Arangasamy A, Venkata KM. Ellagic and ferulic acids protect arsenic-induced male reproductive toxicity via regulating Nfe2l2, Ppargc1a and StAR expressions in testis. Toxicology. 2019;413:1-12. doi:10.1016/j.tox.2018.11.012.
19. Anwar N, Qureshi IZ. In vitro application of sodium arsenite to mice testicular and epididymal organ cultures induces oxidative, biochemical, hormonal, and genotoxic stress. Toxicol Ind Health. 2019;35(10):660-669. doi:10.1177/0748233719885574.
20. Takooree H, Aumeeruddy MZ, Rengasamy KRR, Venugopala KN, Jeewon R, Zengin G, et al. A systematic review on black pepper (Piper nigrum L.): from folk uses to pharmacological applications. Crit Rev Food Sci Nutr. 2019;59(Suppl 1):S210-S243. doi:10.1080/10408398.2019.1565489.
21. Rasve VR, Paithankar VV, Shirsat MK, Dhobale AV. Evaluation of antiulcer activity of Aconitum heterophyllum on experimental animals. World J Pharm Pharm Sci. 2018;7(2):819-839.
22. Shityakov S, Bigdelian E, Hussein AA, Hussain MB, Tripathi YC, Khan MU, et al. Phytochemical and pharmacological attributes of piperine: a bioactive ingredient of black pepper. Eur J Med Chem. 2019;176:149-161. doi:10.1016/j.ejmech.2019.04.002.
23. Dutta S, Majzoub A, Agarwal A. Oxidative stress and sperm function: a systematic review on evaluation and management. Asian J Androl. 2019;17(2):87-97. doi:10.1080/2090598X.2019.1599624.
24. Alahmar AT. Role of oxidative stress in male infertility: an updated review. J Hum Reprod Sci. 2019;12(1):4-18. doi:10.4103/jhrs.JHRS_150_18.
25. Barati E, Nikzad H, Karimian M. Oxidative stress and male infertility: current knowledge of pathophysiology and role of antioxidant therapy in disease management. Cell Mol Life Sci. 2020;77(1):93-113. doi:10.1007/s00018-019-03253-8.
26. Ribas-Maynou J, Yeste M. Oxidative stress in male infertility: causes, effects in assisted reproductive techniques, and protective support of antioxidants. Biology (Basel). 2020;9(4):77. doi:10.3390/biology9040077.
27. Anwar N, Qureshi IZ, Spears N, Lopes F. In vitro administration of sodium arsenite in mouse prepubertal testis induces germ cell loss and apoptosis. Toxicol In Vitro. 2020;67:104924. doi:10.1016/j.tiv.2020.104924.
28. Jaisin Y, Ratanachamnong P, Wongsawatkul O, Watthammawut A, Malaniyom K, Natewong S. Antioxidant and anti-inflammatory effects of piperine on UV-B-irradiated human HaCaT keratinocyte cells. Life Sci. 2020;263:118607. doi:10.1016/j.lfs.2020.118607.
29. Gibb Z, Griffin RA, Aitken RJ, De Iuliis GN. Functions and effects of reactive oxygen species in male fertility. Anim Reprod Sci. 2020;220:106456. doi:10.1016/j.anireprosci.2020.106456.
30. Zhang XY, Wang B, Xu S, Wang J, Gao L, Song YP, et al. Reactive oxygen species-evoked genotoxic stress mediates arsenic-induced suppression of male germ cell proliferation and decline in sperm quality. J Hazard Mater. 2021;406:124768. doi:10.1016/j.jhazmat.2020.124768.
31. Araújo-Ramos AT, Basso CG, Passoni MT, Ribeiro DCK, Spercoski KM, Oliveira JM, et al. The endocrine disrupting effects of sodium arsenite in the rat testis is not mediated through macrophage activation. Reprod Toxicol. 2021;102:1-9. doi:10.1016/j.reprotox.2021.03.005.
32. Haq IU, Imran M, Nadeem M, Tufail T, Gondal TA, Mubarak MS. Piperine: a review of its biological effects. Phytother Res. 2021;35(2):680-700. doi:10.1002/ptr.6855.
33. Zahin M, Bokhari NA, Ahmad I, Husain FM, Althubiani AS, Alruways MW, et al. Antioxidant, antibacterial, and antimutagenic activity of Piper nigrum seeds extracts. Saudi J Biol Sci. 2021;28(9):5094-5105. doi:10.1016/j.sjbs.2021.05.030.
34. Asadi A, Ghahremani R, Abdolmaleki A, Rajaei F. Role of sperm apoptosis and oxidative stress in male infertility: a narrative review. Int J Reprod Biomed. 2021;19(6):493-504. doi:10.18502/ijrm.v19i6.9371.
35. Evans EPP, Scholten JTM, Mzyk A, Reyes-San-Martin C, Llumbet AE, Hamoh T, et al. Male subfertility and oxidative stress. Redox Biol. 2021;46:102071. doi:10.1016/j.redox.2021.102071.
36. Zhang C, Zhao J, Famous E, Pan S, Peng X, Tian J. Antioxidant, hepatoprotective and antifungal activities of black pepper (Piper nigrum L.) essential oil. Food Chem. 2021;346:128845. doi:10.1016/j.foodchem.2020.128845.
37. Rachamalla M, Chinthada J, Kushwaha S, Putnala SK, Sahu C, Jena G, et al. Contemporary comprehensive review on arsenic-induced male reproductive toxicity and mechanisms of phytonutrient intervention. Toxics. 2022;10(12):744. doi:10.3390/toxics10120744.
38. Zargari F, Rahaman MS, KazemPour R, Hajirostamlou M, et al. Arsenic, oxidative stress and reproductive system. J Xenobiot. 2022;12(3):214-222. doi:10.3390/jox12030016.
39. Tripathi AK, Ray AK, Mishra SK. Molecular and pharmacological aspects of piperine as a potential molecule for disease prevention and management: evidence from clinical trials. Beni Suef Univ J Basic Appl Sci. 2022;11:16. doi:10.1186/s43088-022-00196-1.
40. Imran M, Samal M, Qadir A, Ali A, Mir SR. A critical review on the extraction and pharmacotherapeutic activity of piperine. Polim Med. 2022;52(1):31-36. doi:10.17219/pim/145512.
41. Al-Khayri JM, Upadhya V, Pai SR, Naik PM, Al-Mssallem Q, Alessa FM. Comparative quantification of the phenolic compounds, piperine content, and total polyphenols along with the antioxidant activities in the Piper trichostachyon and P. nigrum. Molecules. 2022;27(18):5965. doi:10.3390/molecules27185965.
42. Machado-Neves M. Arsenic exposure and its implications in male fertility. Anim Reprod. 2023;19(4):e20220119. doi:10.1590/1984-3143-AR2022-0119.
43. Peng Z, Yang Q, Yeerken R, Chen J, Liu X, Li X. Multi-omics analyses reveal the mechanisms of arsenic-induced male reproductive toxicity in mice. J Hazard Mater. 2022;424(Pt C):127548. doi:10.1016/j.jhazmat.2021.127548.
44. Li X, Shen K, Yuan D, Fan J, Yang Y, Tian F, et al. Sodium arsenite exposure enhances H3K14 acetylation and impairs male spermatogenesis in rat testes. Reprod Toxicol. 2023;122:108474. doi:10.1016/j.reprotox.2023.108474.
45. Mahmoud MF, Elmaghraby MA, Ali N, Mostafa I, El-Shazly AM, Abdelfattah MAO, et al. Black pepper oil (Piper nigrum L.) mitigates dexamethasone induced pancreatic damage via modulation of oxidative and nitrosative stress. Biomed Pharmacother. 2022;153:113456. doi:10.1016/j.biopha.2022.113456.
46. Yadav SS, Singh MK, Hussain S, Dwivedi P, Khattri S, Singh K. Therapeutic spectrum of piperine for clinical practice: a scoping review. Crit Rev Food Sci Nutr. 2023;63(22):5813-5840. doi:10.1080/10408398.2021.2024792.
47. Yang Y, Hong Y, Han J, Yang Z, Huang N, Xu B, et al. D-Limonene alleviates oxidative stress injury of the testis induced by arsenic in rat. Biol Trace Elem Res. 2024;202(6):2776-2785. doi:10.1007/s12011-023-03881-z.
48. Omar SM, Zahran NN, Alhotan RA, Hussein EO, Galik B, Saleh AA. Evaluation of Salvia hispanica as a therapeutic agent against sodium arsenic-induced testicular toxicity in a male rats model. Life (Basel). 2024;14(1):109. doi:10.3390/life14010109.
49. Eslami H, Ranjbar Askari F, Mahdavi M, Taghavi M, Ghaseminasab-Parizi M. Environmental arsenic exposure and reproductive system toxicity in male and female and mitigatory strategies: a review. Environ Geochem Health. 2024;46(10):420. doi:10.1007/s10653-024-02197-8.
50. Besong EE, et al. Acetate abates arsenic-induced male reproductive toxicity by suppressing HDAC and uric acid-driven oxido-inflammatory NFκB/iNOS/NO response in rats. Biol Trace Elem Res. 2024;202(6):2672-2687. doi:10.1007/s12011-023-03860-4.
51. Mahboob SZ, Jahan S, Badshah H, Virk R, David M, Bano R, et al. Reproductive toxicity in male rats induced by chronic arsenic exposure involves hormonal and structural changes. Sci Rep. 2025;15(1):29037. doi:10.1038/s41598-025-14929-6.