Alpha-pinene alleviates CCl4-induced renal and testicular injury in rats by targeting oxidative stress, inflammation, and apoptosis

Document Type : Original Article

Authors

1 Department of Physiology, Zanjan Branch, Islamic Azad University, Zanjan, Iran

2 Department of Biology, Zanjan Branch, Islamic Azad University, Zanjan, Iran

3 Nanobiotechnology Research Center, Zanjan Branch, Islamic Azad University, Zanjan, Iran

Abstract

Objective(s): Renal and testicular disorders are primarily associated with oxidative damage and inflammation. Here, alpha-pinene (a type of monoterpene) was investigated for its effect on oxidative/nitrosative stress and the expression of inflammatory and apoptotic factors in the kidneys and testes of rats treated with CCl4.
Materials and Methods: CCl4 was injected intraperitoneally (IP) at a dose of 2 ml/kg (twice a week for six weeks). Alpha-pinene (50 mg/kg/day, IP) was also treated during the same period. 
Results: CCl4 increased the level of malondialdehyde (P<0.01 in the kidney and P<0.001 in the testis) and nitric oxide (P<0.001 in the kidney and P<0.01 in the testis) and decreased the levels of glutathione (P<0.05) in the kidneys and testicles of rats. CCl4 also reduced the catalase enzyme activity in the kidneys (P<0.05) but did not affect its activity in the testis. In addition, CCl4 enhanced the mRNA expression of TNF-α (P<0.01), nuclear factor-κB (P<0.05), and Bax (P<0.05 in the kidney and P<0.01 in the testis) and decreased the expression of Bcl-2 (P<0.05) in both organs. Alpha-pinene prevented all the mentioned changes, but it did not influence the expression of Bcl-2 in the kidneys of rats receiving CCl4. 
Conclusion: Alpha-pinene may have the potential to prevent renal and testicular diseases by strengthening the antioxidant system in the kidneys and testis, and inhibiting oxidative/nitrosative stress, inflammation, and apoptosis caused by CCl4.

Keywords

Main Subjects


1. Unsal V, Cicek M, Sabancilar İ. Toxicity of carbon tetrachloride, free radicals and role of anti-oxidants. Rev Environ Health 2021; 36: 279-295. 
2. Ranjbar A, Satari M, Mohseni R, Tavilani A, Ghasemi H. Chlorella vulgaris ameliorates testicular toxicity induced by carbon tetrachloride in male rats via modulating oxidative stress. Andrologia 2022; 54: e14495. 
3. Zargar S, Wani TA. Protective role of quercetin in carbon tetrachloride-induced toxicity in rat brain: biochemical, spectrophotometric assays and computational approach. Molecules 2021; 26: 7526. 
4. Focak M, Suljevic D. Ameliorative Effects of Propolis and Royal Jelly against CCl4‐Induced Hepatotoxicity and Nephrotoxicity in Wistar Rats. Chem Biodivers 2023; 20: e202200948. 
5. Barakat H, Alkabeer IA, Althwab SA, Alfheeaid HA, Alhomaid RM, Almujaydil MS, et al. Nephroprotective effect of fennel (Foeniculum vulgare) seeds and their sprouts on CCl4-induced nephrotoxicity and oxidative stress in rats. Antioxidants 2023; 12: 325-345. 
6. Keshtmand Z, Akbaribazm M, Bagheri Y, Oliaei R. The ameliorative effects of Lactobacillus coagulans and Lactobacillus casei probiotics on CCl4‐induced testicular toxicity based on biochemical, histological and molecular analyses in rat. Andrologia 2021; 53: e13908. 
7. He S, Zhao W, Chen X, Li J, Zhang L, Jin H. Ameliorative effects of peptide phe-leu-ala-pro on acute liver and kidney injury caused by CCl4 via attenuation of oxidative stress and inflammation. ACS omega 2022; 7: 44796-44803. 
8. Abdel Moneim AE. Prevention of carbon tetrachloride (CCl4)-induced toxicity in testes of rats treated with Physalis peruviana L. fruit. Toxicol Ind Health 2016; 32: 1064-1073. 
9. Hwang IS, Kim JE, Lee YJ, Kwak MH, Choi YH, Kang BC, et al. Protective effects of gomisin A isolated from Schisandra chinensis against CCl4-induced hepatic and renal injury. International J Mol Med 2013; 31: 888-898. 
10. Ammar NM, Hassan HA, Abdallah HM, Afifi SM, Elgamal AM, Farrag AR, et al. Protective effects of naringenin from Citrus sinensis (var. Valencia) peels against CCl4-induced hepatic and renal injuries in rats assessed by metabolomics, histological and biochemical analyses. Nutr 2022; 14: 841. 
11. Baccouri B, Rajhi I. Potential anti-oxidant activity of terpenes. In Perveen S, Mohammed Al-Taweel A editor. Terpenes and Terpenoids-Recent Advances. IntechOpen; 2021.p. 53-62. 
12. Aydin E, Türkez H, Geyikoğlu F. Anti-oxidative, anticancer and genotoxic properties of α-pinene on N2a neuroblastoma cells. Biologia 2013; 68: 1004-1009. 
13. Rahmani H, Moloudi MR, Hashemi P, Hassanzadeh K, Izadpanah E. Alpha-Pinene alleviates motor activity in animal model of Huntington’s disease via enhancing anti-oxidant capacity. Neurochem Res 2023; 48: 1775–1782 
14. Khan‐Mohammadi‐Khorrami MK, Asle‐Rousta M, Rahnema M, Amini R. Neuroprotective effect of alpha‐pinene is mediated by suppression of the TNF‐α/NF‐κB pathway in Alzheimer’s disease rat model. J Biochem Mol Toxicol 2022; 36: e23006. 
15. Karthikeyan R, Kanimozhi G, Prasad NR, Agilan B, Ganesan M, Srithar G. Alpha pinene modulates UVA-induced oxidative stress, DNA damage and apoptosis in human skin epidermal keratinocytes. Life Sci 2018; 212: 150- 158. 
16. Hashemi P, Ahmadi S. Alpha-pinene exerts antiseizure effects by preventing oxidative stress and apoptosis in the hippocampus in a rat model of temporal lobe epilepsy induced by kainate. Mol Neurobiol 2023; 60: 3227-3238.
17. Santos ES, de Sousa Machado ST, Rodrigues FB, da Silva YA, Matias LC, Lopes MJ, et al. Potential anti-inflammatory, hypoglycemic, and hypolipidemic activities of alpha-pinene in diabetic rats. Process Biochem 2023; 126: 80-86. 
18. Zhang B, Wang H, Yang Z, Cao M, Wang K, Wang G, et al. Protective effect of alpha-pinene against isoproterenol-induced myocardial infarction through NF-κB signaling pathway. Hum Exp Toxicol 2020; 39: 1596-1606. 
19. Khoshnazar M, Bigdeli MR, Parvardeh S, Pouriran R. Attenuating effect of α-pinene on neurobehavioural deficit, oxidative damage and inflammatory response following focal ischaemic stroke in rat. J Pharm Pharmacol 2019; 71: 1725-1733. 
20. Wang R, Song F, Li S, Wu B, Gu Y, Yuan Y. Salvianolic acid A attenuates CCl4-induced liver fibrosis by regulating the PI3K/AKT/mTOR, Bcl-2/Bax and caspase-3/cleaved caspase-3 signaling pathways. Drug Des Devel Ther 2019: 1889-1900. 
21. Aydın MM, Akçalı KC. Liver fibrosis. Turk J Gastroenterol 2018; 29: 14-21. 
22. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193: 265-275. 
23. Draper HH, Hadley M. Malondialdehyde determination as index of lipid Peroxidation. In Methods in enzymology. Academic press; 1990.p. 421-431. 
24. Iverson NM, Hofferber EM, Stapleton JA. Nitric oxide sensors for biological applications. Chemosensors 2018; 6: 8. 
25. Jollow DJ, Mitchell JR, Zampaglione NA, Gillette JR. Bromobenzene-induced liver necrosis. Protective role of glutathione and evidence for 3, 4-bromobenzene oxide as the hepatotoxic metabolite. Pharmacology 1974; 11: 151-169. 
26. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 2001; 25: 402-408. 
27. Türk G, Çeribaşı S, Sönmez M, Çiftçi M, Yüce A, Güvenç M, et al. Ameliorating effect of pomegranate juice consumption on carbon tetrachloride-induced sperm damages, lipid peroxidation, and testicular apoptosis. Toxicol Ind Health 2016; 32: 126-137. 
28. Abd-Elhakim YM, Ghoneim MH, Ebraheim LL, Imam TS. Taurine and hesperidin rescues carbon tetrachloride-triggered testicular and kidney damage in rats via modulating oxidative stress and inflammation. Life Sci 2020; 254: 117782. 
29. Hashem AS. Defensive impact of propolis against CCl4 actuated rats’ testicular damage. J Adv Vet Anim Res 2021; 8: 70-77. 
30. Abdel Moneim AE. Prevention of carbon tetrachloride (CCl4)-induced toxicity in testes of rats treated with Physalis peruviana L. fruit. Toxicol Ind Health 2016; 32: 1064-1073. 
31. Tucker PS, Scanlan AT, Dalbo VJ. Chronic kidney disease influences multiple systems: describing the relationship between oxidative stress, inflammation, kidney damage, and concomitant disease. Oxid Med Cell Longev 2015; 2015. 
32. Dutta S, Sengupta P, Slama P, Roychoudhury S. Oxidative stress, testicular inflammatory pathways, and male reproduction. Int J Mol Sci 2021; 22: 10043. 
33. Liu T, Zhang L, Joo D, Sun SC. NF-κB signaling in inflammation. Signal Transduct Target Ther 2017; 2: 17023. 
34. Kim DS, Lee HJ, Jeon YD, Han YH, Kee JY, Kim HJ, et al. Alpha-pinene exhibits anti-inflammatory activity through the suppression of MAPKs and the NF-κB pathway in mouse peritoneal macrophages. Am J Chinese Med 2015; 43: 731-742. 
35. Aktan F. iNOS-mediated nitric oxide production and its regulation. Life Sci 2004; 75: 639-653. 
36. Al-Shabanah OA, Alam K, Nagi MN, Al-Rikabi AC, Al-Bekairi AM. Protective effect of aminoguanidine, a nitric oxide synthase inhibitor, against carbon tetrachloride induced hepatotoxicity in mice. Life Sci 1999; 66: 265-270. 
37. Chandra J, Samali A, Orrenius S. Triggering and modulation of apoptosis by oxidative stress. Free Radic Biol Med 2000; 29: 323-333. 
38. Van den Berg R, Haenen GR, Van den Berg H, Bast A. Transcription factor NF-κB as a potential biomarker for oxidative stress. Br J Nutr 2001; 86: S121- S127. 
39. Pentikäinen V, Suomalainen L, Erkkilä K, Martelin E, Parvinen M, Pentikäinen MO, Dunkel L. Nuclear factor- κB activation in human testicular apoptosis. Am J Pathol 2002; 160: 205-218. 
40. White S, Lin L, Hu K. NF-κB and tPA signaling in kidney and other diseases. Cells 2020; 9: 1348. 
41. Ma JQ, Ding J, Zhang L, Liu CM. Hepatoprotective properties of sesamin against CCl4 induced oxidative stress-mediated apoptosis in mice via JNK pathway. Food Chem Toxicol 2014; 64: 41-48. 
42. Raji-Amirhasani A, Khaksari M, Soltani Z, Saberi S, Iranpour M, Darvishzadeh Mahani F, et al. Beneficial effects of time and energy restriction diets on the development of experimental acute kidney injury in Rat: Bax/Bcl-2 and histopathological evaluation. BMC nephrol 2023; 24: 59-71. 
43. Rehman MU, Tahir M, Khan AQ, Khan R, Oday-O-Hamiza, Lateef A, et al. D-limonene suppresses doxorubicin-induced oxidative stress and inflammation via repression of COX-2, iNOS, and NFκB in kidneys of Wistar rats. Exp Biol Med 2014; 239: 465-476. 
44. Shereen MS, Mahmoud A, Mohamed ZB, Mohamed A. Effect of D-limonene on the age-related androgenic changes in male rats. Med J Cairo Uni 2020; 88: 599-609. 
45. Said MM. The protective effect of eugenol against gentamicin‐induced nephrotoxicity and oxidative damage in rat kidney. Fundam Clin Pharmacol 2011; 25: 708-716. 
46. Ekinci Akdemir FN, Yildirim S, Kandemir FM, Aksu EH, Guler MC, Kiziltunc Ozmen H, et al. The antiapoptotic and anti-oxidant effects of eugenol against cisplatin‐induced testicular damage in the experimental model. Andrologia 2019; 51: e13353. 
47. Dayangac A, Bahsi M, Ozkaya A, Yilmaz O. Linalool improve biochemical damage and fatty acids composition of testes on fasting male rats. J Anim Vet Adv 2011; 10: 1232- 1238. 
48. Altinoz E, Oner Z, Elbe H, Uremis N, Uremis M. Linalool exhibits therapeutic and protective effects in a rat model of doxorubicin-induced kidney injury by modulating oxidative stress. Drug Chem Toxicol 2022; 45: 2024-2030. 
49. Lin TC, Lu CW, Chang KF, Lee CJ. Juniperus communis extract ameliorates lipopolysaccharide‐induced acute kidney injury through the adenosine monophosphate–activated protein kinase pathway. Food Sci Nutr 2022; 10: 3405-3414. 
50. Saied M, Ali K, Mosayeb A. Rosemary (Rosmarinus officinalis L.) essential oil alleviates testis failure induced by etoposide in male rats. Tissue Cell. 2023: 102016.