Attenuation of tacrolimus induced oxidative stress, mitochondrial damage, and cell cycle arrest by Boerhavia diffusa root fraction in mdck cell lines

Document Type : Original Article

Authors

1 Department of Biomedical Sciences, Sri Ramachandra Institute of Higher Education and Research (DU), Porur, Chennai 6000116, India

2 Department of Human Genetics, Sri Ramachandra Institute of Higher Education and Research (DU), Porur, Chennai, 600116, India

Abstract

Objective(s): The protective effect of ethyl acetate fraction (EAF) of Boerhavia diffusa roots against Tacrolimus (TAC) induced nephrotoxicity was studied using MDCK cell lines.
Materials and Methods: Ethanolic root extract of B. diffusa was fractionated using the liquid-liquid partition method. The cytotoxic effect of TAC and protective effect of EAF co-treatment were studied in MDCK cell lines by measuring ROS, LPO, and NO levels; collagen accumulation, effect on mitochondrial membrane integrity and cell cycle analysis were studied. The active component in EAF was quantified by HPLC analysis.
Results: TAC induced toxicity, leading to apoptosis and necrosis, was significantly reduced (p <0.001) in EAF co-treatment, with reversal of cell cycle arrest and reduced cell population at sub G0/G1 phase. Further, ROS (p <0.05), LPO and NO (p <0.001), were significantly reduced with EAF co-treatment compared with TAC individually treated cells. TAC induced mitochondrial membrane integrity loss was found to be significantly reduced in co-treated cells, as measured by rhodamine123 (p <0.05) and translocation of cytochrome c (p <0.001) from nucleus to cytoplasm, and caspase 3 release (p <0.001). The same was confirmed through annexin-FITC and PI staining (p <0.05) with reduced apoptotic and necrotic death in co-treated population. Interestingly, EAF co-treatment decreased collagen accumulation (p <0.001) with significant increase in the cell survival of tubular epithelial cells. HPLC analysis showed the presence of Quercetin (87.5 mg/g) in EAF, which may be responsible for the nephroprotective role.
Conclusion: Thus, these results provide sound evidence that EAF may be an effective adjuvant therapy to prevent nephrotoxicity induced by TAC.

Keywords

References

1. Naesens M, Kuypers DR, Sarwal M. Calcineurin inhibitor nephrotoxicity. Clin J Am Soc Nephrol 2009; 4:481-508.
2. Zhou X, Yang G, Davis CA, Doi SQ, Hirszel P, Wingo CS, et al. Hydrogen peroxide mediates FK506-induced cytotoxicity in renal cells. Kidney Int 2004; 65:139-147.
3. Matas AJ, Smith JM, Skeans MA, Lamb KE, Gustafson SK, Samana CJ, et al. OPTN/SRTR 2011 annual data report: Kidney. Am J Transplant 2013; 13 Suppl 1:11-46.
4. Hošková L, Málek I, Kautzner J, Honsová E, van Dokkum RPE, Husková Z, et al. Tacrolimus-induced hypertension and nephrotoxicity in Fawn-Hooded rats are attenuated by dual inhibition of renin–angiotensin system. Hypertens Res 2014; 37:724-732.
5. Khanna AK, Pieper GM. NADPH oxidase subunits (NOX-1, p22phox, Rac-1) and tacrolimus-induced nephrotoxicity in a rat renal transplant model. Nephrol Dial Transplant 2006; 22:376-385.
6. Ara C, Dirican A, Unal B, Bay Karabulut A, Piskin T. The effect of melatonin against fk506-induced renal oxidative stress in rats. Surg Innov 2011; 18:34-38.
7. Hisamura F, Kojima-Yuasa A, Kennedy DO, Matsui-Yuasa I. Protective effect of green tea extract and tea polyphenols against fk506-induced cytotoxicity in renal cells. Basic Clin Pharmacol Toxicol 2006; 98:192-196.
8. Li X, Zhuang S. Recent advances in renal interstitial fibrosis and tubular atrophy after kidney transplantation. Fibrogenesis & Tissue Repair 2014; 7:1-11.
9. Butani L, Afshinnik A, Johnson J, Javaheri D, Peck S, German JB, et al. Amelioration of tacrolimus-induced nephrotoxicity in rats using juniper oil. Transplantation 2003; 76:306-311.
10. Suman S, Hayagreeva Dinakar Y, Suhas reddy P V, Sai Sudha Yadav B, Venkateshwar Reddy V. Ameliorative effect of Cubeba Officinalis dried fruits against Tacrolimus induced nephrotoxicity in Wistar albino rats.Int. J Res Pharm Sci 2020; 11:596-602.
11. Oyouni A, Saggu S, Tousson E, Mohan A, Farasani A. Mitochondrial nephrotoxicity induced by tacrolimus (FK-506) and modulatory effects of Bacopa monnieri (Farafakh) of Tabuk Region. Pharmacognosy Res 2019; 11:20-24.
12. Oyouni AAA, Saggu S, Tousson E, Rehman H. Immunosuppressant drug tacrolimus induced mitochondrial nephrotoxicity, modified PCNA and Bcl-2 expression attenuated by Ocimum basilicum L. in CD1 mice. Toxicol Rep 2018; 5:687-694.
13. Jegan N, editor. Effect of Hydroalcoholic Extract of Boerhaavia Diffusa Linn against Cisplatin Induced Nephrotoxicity2015; 10:17-22.
14. Padmini P. An experimental study of biochemical and histopathological study on gentamycin induced renal failure in albino rat and the effectiveness of punarnava (Boerhaevia Diffusa) on reversal of renal damage. IOSR JDMS 2013; 9:17-21.
15. Sawardekar S, Patel T. Evaluation of the effects of boerhaavia diffusa on gentamicin induced nephrotoxicity in rats. J Ayurveda Integr Med 2015; 6:95-103.
16. Yadav HN, Sharma US, Singh S, Gupta YK. Effect of combination of Tribulus terrestris, Boerhavia diffusa and Terminalia chebula reverses mercuric chloride-induced nephrotoxicity and renal accumulation of mercury in rat. Orient Pharm Exp Med 2019; 19:497-507.
17. Karwasra R, Kalra P, Nag T, Gupta Y, Singh S, Panwar A. Safety assessment and attenuation of cisplatin induced nephrotoxicity by tuberous roots of Boerhaavia diffusa. Regul Toxicol Pharm 2016; 81:341-352.
18. Indhumathi T, Shipa K, Mohandass S. Evaluation of nephroprotective role of Boerhaavea diffusa leaves against mercuric chloride induced toxicity in experimental rats. J Pharm Res 2011; 4:1848-1850.
19. M.K K, Soundararajan P, Vasanthi HR, Arockiasamy S. In vitro nephroprotective role of ethanolic root extract of boerhaavia diffusa against cisplatin induced nephrotoxicity. Int J Phytomed 2015;7: 388-395.
20. Bowlekar PB, Gadgoli CH. Amelioration of cisplatin induced nephrotoxicity by standardized methanolic extract of roots of boerhaavia diffusa. Nat Prod J 2014; 4:217-223.
21. Kalaivani M, Arockiasamy S, John C, Vasanthi H, Soundararajan P. Therapeutic potential of Boerhavia diffusa L. against cyclosporine A-Induced mitochondrial dysfunction and apoptosis in madin–Darby canine kidney cells. Pharmacogn Mag 2018; 14:132-140.
22. Erdogan S, Doganlar O, Doganlar ZB, Turkekul K. Naringin sensitizes human prostate cancer cells to paclitaxel therapy. Prostate Int 2018; 6:126-135.
23. Repetto G, del Peso A, Zurita JL. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nat Protoc 2008; 3:1125-1131.
24. Elumalai P, Gunadharini DN, Senthilkumar K, Banudevi S, Arunkumar R, Benson CS, et al. Induction of apoptosis in human breast cancer cells by nimbolide through extrinsic and intrinsic pathway. Toxicol Lett 2012; 215:131-142.
25. Servais H, Ortiz A, Devuyst O, Denamur S, Tulkens PM, Mingeot-Leclercq MP. Renal cell apoptosis induced by nephrotoxic drugs: cellular and molecular mechanisms and potential approaches to modulation. Apoptosis 2008; 13:11-32.
26. Ligasová A, Koberna K. Quantification of fixed adherent cells using a strong enhancer of the fluorescence of DNA dyes. Sci. Rep 2019; 9:8701.
27.  Balaji C, Muthukumaran J, Vinothkumar R, Nalini N. Anticancer effects of sinapic acid on human colon cancer cell lines HT-29 and SW480. Int J Pharm Biol Arch 2014; 5: 176-183.
28. Hsieh C-Y, Chen C-L, Yang K-C, Ma C-T, Choi P-C, Lin C-F. Detection of reactive oxygen species during the cell cycle under normal culture conditions using a modified fixed-sample staining method. Immunoassay Immunochem 2015; 36:149-161.
29. Filipkowski RK, Hetman M, Kaminka B, Kaczmarek L. DNA fragmentation in rat brain after intraperitoneal administration of kainate. Neuroreport 1994; 5:1538-1540.
30. Satoh M, Kashihara N, Fujimoto S, Horike H, Tokura T, Namikoshi T, et al. A novel free radical scavenger, edarabone, protects against cisplatin-induced acute renal damage in vitro and in vivo. J Pharmacol Exp Ther 2003; 305:1183-1190.
31. Hortelano S, López-Collazo E, Boscá L. Protective effect of cyclosporin A and FK506 from nitric oxide-dependent apoptosis in activated macrophages. Br J Pharmacol 1999; 126:1139-1146.
32. Taso OV, Philippou A, Moustogiannis A, Zevolis E, Koutsilieris M. Lipid peroxidation products and their role in neurodegenerative diseases. Ann Res Hosp 2019; 3:2-10.
33. Zhong Z, Connor HD, Li X, Mason RP, Forman DT, Lemasters JJ, et al. Reduction of ciclosporin and tacrolimus nephrotoxicity by plant polyphenols. J Pharm Pharmacol 2006; 58:1533-1543.
34. Mishra S, Aeri V, Gaur PK, Jachak SM. Phytochemical, therapeutic, and ethnopharmacological overview for a traditionally important herb: Boerhavia diffusa linn. BioMed Research International 2014; 2014:1-19.
35. Ortiz A, Lorz C, Catalán M, Ortiz A, Coca S, Egido J. Cyclosporine A induces apoptosis in murine tubular epithelial cells: Role of caspases. Kidney Int 1998; 54:S25-S29.
36. Ali AS, Almalki AS, Alharthy BT. Effect of kaempferol on tacrolimus-induced nephrotoxicity and calcineurin B1 expression level in animal model. J Exp Pharmacol 2020; 12:397-407.
37. Park C, Kwon DH, Hwang SJ, Han MH, Jeong J-W, Hong SH, et al. Protective effects of nargenicin a1 against tacrolimus-induced oxidative stress in hirame natural embryo cells. International journal of environmental research and public health 2019; 16:1-13.
38. Kim ES, Lee JS, Akram M, Kim KA, Shin YJ, Yu JH, et al. Protective activity of dendropanax morbifera against cisplatin-induced acute kidney injury. Kidney Blood Press. Res 2015; 40:1-12.
39. Farris AB, Colvin RB. Renal interstitial fibrosis: Mechanisms and evaluation in: Current opinion in nephrology and hypertension. Curr Opin Nephrol Hypertens 2012; 21:289-300.
40. Zhu Q, Hu J, Meng H, Shen Y, Zhou J, Zhu Z. S-Phase cell cycle arrest, apoptosis, and molecular mechanisms of aplasia ras homolog member i–induced human ovarian cancer skov3 cell lines. Int J Gynecol Cancer 2014; 24:629-634.
41. Prathapan A, Vineetha VP, Raghu KG. Protective effect of Boerhaavia diffusa L. against mitochondrial dysfunction in angiotensin II induced hypertrophy in H9c2 cardiomyoblast cells. PLoS One 2014; 9:e96220.
42. Chaudhary S, Ganjoo P, Raiusddin S, Parvez S. Nephroprotective activities of quercetin with potential relevance to oxidative stress induced by valproic acid. Protoplasma 2015; 252:209-217.
43. Lim SW, Shin YJ, Luo K, Quan Y, Cui S, Ko EJ, et al. Ginseng increases Klotho expression by FoxO3-mediated manganese superoxide dismutase in a mouse model of tacrolimus-induced renal injury. Aging 2019; 11:5548-5569.
44. Abdel-Aziz A-A, El-Din M, Balah A, Akool E-S. Molecular mechanisms of the modulatory effect of vitamin E on tacrolimus (FK506)-induced renal injury in rats. Br J Pharm Res 2016; 9:1-9.
45. Wongmekiat O, Leelarugrayub N, Thamprasert K. Beneficial effect of shallot (Allium ascalonicum L.) extract on cyclosporine nephrotoxicity in rats. Food and chemical toxicology: An international journal published for the British Industrial Biological Research Association 2008; 46:1844-1850.
46. Hisamura F, Kojima-Yuasa A, Huang X, Kennedy DO, Matsui-Yuasa I. Synergistic effect of green tea polyphenols on their protection against FK506-induced cytotoxicity in renal cells. Am J Chin Med 2008; 36:615-624.
47. Jeon SH, Park HM, Kim SJ, Lee MY, Kim GB, Rahman MM, et al. Taurine reduces FK506-induced generation of ROS and activation of JNK and Bax in Madin Darby canine kidney cells. Hum Exp Toxicol 2010; 29:627-633.
48. Mehrotra S, Mishra KP, Maurya R, Srimal RC, Singh VK. Immunomodulation by ethanolic extract of Boerhaavia diffusa roots. Int Immunopharmacol 2002; 2:987-996.
49. Athira KV, Madhana RM, Lahkar M. Flavonoids, the emerging dietary supplement against cisplatin-induced nephrotoxicity. Chem Biol Interact 2016; 248:18-20.
50. Vargas F, Romecín P, García-Guillén AI, Wangesteen R, Vargas-Tendero P, Paredes MD, et al. Flavonoids in kidney health and disease. Front Physiol 2018; 9:1-12.
51. Zal F, Mostafavi-Pour Z, Vessal M. Comparison of the effects of vitamin E and/or quercetin in attenuating chronic cyclosporine A-induced nephrotoxicity in male rats. Clin Exp Pharmacol Physiol 2007; 34:720-724.
Iranian Journal of Basic Medical Sciences
Volume 24, Issue 8
August 2021
Pages 1087-1097

Files

Share

How to cite

Statistics