JNK and p38 gene and protein expression during liver ischemia-reperfusion in a rat model treated with silibinin

Document Type : Original Article

Authors

1 Department of Clinical Biochemistry and Genetics, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran

2 Genetic Biology, Islamic Azad University of Tonekabon, Tonekabon, Iran

3 Department of Anatomy and Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran

4 Faculty of Medicine, Immunogenetic Research Center (IRC), Mazandaran University of Medical Sciences, Sari, Iran

Abstract

Objective(s): Signal transduction of mitogen-activated protein kinases (MAPKs) is activated during ischemia. In this study, c-Jun N-terminal Kinase (JNK) and p38 MAPK (p38) gene and protein expression were evaluated as two members of the MAPK family during liver ischemia-reperfusion in rats.
Materials and Methods: Thirty-two male Wistar rats were divided into four groups of eight: Vehicle, ischemia-reperfusion (IR), ischemia-reperfusion+silibinin (IR+SILI), and SILI. The IR and IR+SILI groups differed from the other two groups in that they underwent one hour of ischemia followed by three hr of reperfusion. The Vehicle and IR groups received normal saline while the SILI and IR+SILI groups were treated with silibinin (50 mg/kg). At the end of the reperfusion time, blood and ischemic liver tissue were collected for further experiments.
Results: The expression of JNK and p38 gene, the amount of serum hepatic injury indices, and malondialdehyde (MDA) in the IR group increased significantly compared with the vehicle group. The JNK and p38 gene expression decreased significantly in the IR + SILI group compared with the IR group. Glutathione peroxidase (GPx) and total antioxidant capacity (TAC) levels decreased in the IR group while increasing in the IR+SILI group. Histological examination showed that silibinin significantly reduced the severity of hepatocyte degradation. Western blot results were completely consistent with real-time PCR results.
Conclusion: The possible pathways of the protective effect of silibinin against hepatic ischemia damages is to reduce the expression of the p38 and JNK gene and protein. 

Keywords

References

1. Saidi RF, Kenari SKH. Liver ischemia/reperfusion injury: An overview. J Invest Surg 2014;27:366-379.
2. Xie J, Guo Q. Apoptosis antagonizing transcription factor protects renal tubule cells against oxidative damage and apoptosis induced by ischemia-reperfusion. J Am Soc Nephrol 2006;17:3336-3346.
3. Feng J, Zhang Q, Mo W, Wu L, Li S, Li J, et al. Salidroside pretreatment attenuates apoptosis and autophagy during hepatic ischemia–reperfusion injury by inhibiting the mitogen-activated protein kinase pathway in mice. Drug Des Devel Ther 2017;11:1989-2006.
4. Du X, Shi Z, Peng Z, Zhao C, Zhang Y, Wang Z, et al. Acetoacetate induces hepatocytes apoptosis by the ROS‐mediated MAPKs pathway in ketotic cows. J Cell Physiol 2017;232:3296-3308.
5. Oltean M. Silybin Against Liver Ischemia-Reperfusion Injury: Something Old, Something New. J Invest Surg 2018;31:521-522.
6. Qajari NM, Shafaroudi MM, Gholami M, Khonakdar-Tarsi A. Silibinin treatment results in reducing OPA1&MFN1 genes expression in a rat model hepatic ischemia-reperfusion. Mol Biol Rep 2020;47:3271-3280.
7. Rao X, Huang X, Zhou Z, Lin X. An improvement of the 2ˆ (–delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath 2013;3:71-85.
8. Serracino-Inglott F, Habib NA, Mathie RT. Hepatic ischemia-reperfusion injury. Am J Surg 2001;181:160-166.
9. Jaeschke H, Farhood A, Smith CW. Neutrophils contribute to ischemia/reperfusion injury in rat liver in vivo. FASEB J 1990;4:3355-3359.
10. Vascotto C, Cesaratto L, D’Ambrosio C, Scaloni A, Avellini C, Paron I, et al. Proteomic analysis of liver tissues subjected to early ischemia/reperfusion injury during human orthotopic liver transplantation. Proteomics. 2006;6:3455-3465.
11. Grace P. Ischaemia‐reperfusion injury. Br J Surg 1994;81:637-647.
12. Chouchani ET, Pell VR, Gaude E, Aksentijević D, Sundier SY, Robb EL, et al. Ischaemic accumulation of succinate controls reperfusion injury through mitochondrial ROS. Nature 2014;515:431-435.
13. Zhang C, Xu X, Potter BJ, Wang W, Kuo L, Michael L, et al. TNF-α contributes to endothelial dysfunction in ischemia/reperfusion injury. Arterioscler Thromb Vasc Biol 2006;26:475-480.
14. Semenza GL. Cellular and molecular dissection of reperfusion injury: ROS within and without. Circ Res 2000;86:117-118.
15. Takada M, Nadeau KC, Shaw GD, Marquette KA, Tilney NL. The cytokine-adhesion molecule cascade in ischemia/reperfusion injury of the rat kidney. Inhibition by a soluble P-selectin ligand. J Clin Invest 1997;99:2682-2690.
16. Brar BK, Jonassen AK, Stephanou A, Santilli G, Railson J, Knight RA, et al. Urocortin protects against ischemic and reperfusion injury via a MAPK-dependent pathway. J Biol Chem 2000;275:8508-8514.
17. Jeong C-W, Yoo KY, Lee SH, Jeong HJ, Lee CS, Kim SJ. Curcumin protects against regional myocardial ischemia/reperfusion injury through activation of RISK/GSK-3β and inhibition of p38 MAPK and JNK. J Cardiovasc pharmacol Ther 2012;17:387-394.
18. Yoshinari D, Takeyoshi I, Kobayashi M, Koyama T, Iijima K, Ohwada S, et al. Effects of a P38 mitogen-activated protein kinases inhibitor as an additive to university of Wisconsin solution on reperfusion injury in liver transplantation. Transplantation 2001;72:22-27.
19. Ocuin LM, Zeng S, Cavnar MJ, Sorenson EC, Bamboat ZM, Greer JB, et al. Nilotinib protects the murine liver from ischemia/reperfusion injury. J Hepatol 2012;57:766-773.
20. Kang HH, Kim IK, in Lee H, Joo H, Lim JU, Lee J, et al. Chronic intermittent hypoxia induces liver fibrosis in mice with diet-induced obesity via TLR4/MyD88/MAPK/NF-kB signaling pathways. Biochem Biophys Res Commun 2017;490:349-355.
21. King LA, Toledo AH, Rivera-Chavez FA, Toledo-Pereyra LH. Role of p38 and JNK in liver ischemia and reperfusion. J hepatobiliary Pancreat Surg 2009;16:763-770.
22. Dong X, Swaminathan S, Bachman L-A, Croatt A-J, Nath K-A, Griffin M-D. Resident dendritic cells are the predominant TNF-secreting cell in early renal ischemia–reperfusion injury. Kidney Int 2007;71:619-628.
23. Fujimoto H, Ohno M, Ayabe S, Kobayashi H, Ishizaka N, Kimura H, et al. Carbon monoxide protects against cardiac ischemia-reperfusion injury in vivo via MAPK and Akt-eNOS pathways. Arterioscler Thromb Vasc Biol 2004;24:1848-1853.
24. Xu S, Niu P, Chen K, Xia Y, Yu Q, Liu N, et al. The liver protection of propylene glycol alginate sodium sulfate preconditioning against ischemia reperfusion injury: Focusing MAPK pathway activity. Sci Rep 2017;7:1-12.
25. Weston CR, Wong A, Hall JP, Goad ME, Flavell RA, Davis RJ. JNK initiates a cytokine cascade that causes Pax2 expression and closure of the optic fissure. Genes Dev 2003;17:1271-1280.
26. Ozbek E, Cekmen M, Ilbey YO, Simsek A, Polat EC, Somay A. Atorvastatin prevents gentamicin-induced renal damage in rats through the inhibition of p38-MAPK and NF-kB pathways. Ren Fail 2009;31:382-392.
27. Sironi L, Banfi C, Brioschi M, Gelosa P, Guerrini U, Nobili E, et al. Activation of NF-kB and ERK1/2 after permanent focal ischemia is abolished by simvastatin treatment. Neurobiol Dis 2006;22:445-451.
28. Baker SJ, Reddy EP. Transducers of life and death: TNF receptor superfamily and associated proteins. Oncogene 1996;12:1-9.
29. Kim J, Lee S, Park J, Yoo Y. TNF-α-induced ROS production triggering apoptosis is directly linked to Romo1 and Bcl-XL. Cell Death Differ 2010;17:1420-1434.
30. Feng X. Regulatory roles and molecular signaling of TNF family members in osteoclasts. Gene 2005;350:1-13.
31. Jupp OJ, McFarlane SM, Anderson HM, Littlejohn AF, Mohamed AA, MacKay RH, et al. Type II tumour necrosis factor-α receptor (TNFR2) activates c-Jun N-terminal kinase (JNK) but not mitogen-activated protein kinase (MAPK) or p38 MAPK pathways. Biochem J 2001;359:525-535.
32. Liz-Grana M, Gómez-Reino Carnota J. Tumour necrosis factor. Genetics, cell action mechanism and involvement in inflammation. Allergol Immunol Clin 2001;16:140-149.
33. Mandic A. Elucidation of pro-apoptotic signaling induced by cisplatin. Karolinska Institute, Stockholm, 2003.
34. Liang Q, Molkentin JD. Redefining the roles of p38 and JNK signaling in cardiac hypertrophy: dichotomy between cultured myocytes and animal models. J Mol Cell Cardiol 2003;35:1385-1394.
35. Matsuda S, Moriguchi T, Koyasu S, Nishida E. T lymphocyte activation signals for interleukin-2 production involve activation of MKK6-p38 and MKK7-SAPK/JNK signaling pathways sensitive to cyclosporin A. J Biol Chem 1998;273:12378-12382.
36. Gwak G-Y, Moon TG, Lee DH, Yoo BC. Glycyrrhizin attenuates HMGB1-induced hepatocyte apoptosis by inhibiting the p38-dependent mitochondrial pathway. World J Gastroenterol  2012;18:679-684.
37. Cannistrà M, Ruggiero M, Zullo A, Gallelli G, Serafini S, Maria M, et al. Hepatic ischemia reperfusion injury: A systematic review of literature and the role of current drugs and biomarkers. Int J Surg 2016;33:S57-S70.
38. Feng L, Ke N, Cheng F, Guo Y, Li S, Li Q, et al. The protective mechanism of ligustrazine against renal ischemia/reperfusion injury. J Surg Res 2011;166:298-305.
39. Sanches SC, Ramalho LNZ, Mendes-Braz M, Terra VA, Cecchini R, Augusto MJ, et al. Riboflavin (vitamin B-2) reduces hepatocellular injury following liver ischaemia and reperfusion in mice. Food Chem Toxicol 2014;67:65-71.
40. Zhong Z, Froh M, Connor HD, Li X, Conzelmann LO, Mason RP, et al. Prevention of hepatic ischemia-reperfusion injury by green tea extract. Am J Physiol Gastrointest Liver Physiol 2002;283:G957-G964.
41. Flora K, Hahn M, Rosen H, Benner K. Milk thistle (Silybum marianum) for the therapy of liver disease. Am J Gastroenterol 1998;93:139-143.
42. Liu S, Chen Z, Cao D, Liu F, Wang X, Zhao L, et al. Study on protective effect of extract on hepatic ischemia-reperfusion injury. Afr J Tradit Complement Altern Med 2013;10:310-312.
43. Post-White J, Ladas EJ, Kelly KM. Advances in the use of milk thistle (Silybum marianum). Integr Cancer Ther 2007;6:104-109.
44. Freitag AF, Cardia GFE, da Rocha BA, Aguiar RP, Silva-Comar FMdS, Spironello RA, et al. Hepatoprotective effect of silymarin (Silybum marianum) on hepatotoxicity induced by acetaminophen in spontaneously hypertensive rats. Evid Based Complement Alternat Med 2015;2015:538317.
45. Çağlayan F, Çağlayan O, Gunel E, Elcuman Y, Çakmak M. Intestinal ischemia-reperfusion and plasma enzyme levels. Pediatr Surg Int 2002;18:255-257.
46. Yuan G-J, Ma J-C, Gong Z-J, Sun X-M, Zheng S-H, Li X. Modulation of liver oxidant-antioxidant system by ischemic preconditioning during ischemia/reperfusion injury in rats. World J Gastroenterol 2005;11:1825.
47. Sewerynek E, Reiter RJ, Melchiorri D, Ortiz GG, Lewinski A. Oxidative damage in the liver induced by ischemia-reperfusion: Protection by melatonin. Hepatogastroenterology 1996;43:898-905.
48. Czubkowski P, Socha P, Pawlowska J. Current status of oxidative stress in pediatric liver transplantation. Pediatr Transplant 2010;14:169-177.
49. Senturk H, Kabay S, Bayramoglu G, Ozden H, Yaylak F, Yucel M, et al. Silymarin attenuates the renal ischemia/reperfusion injury-induced morphological changes in the rat kidney. World J Urol 2008;26:401-407.
50. Chen P-N, Hsieh Y-S, Chiou H-L, Chu S-C. Silibinin inhibits cell invasion through inactivation of both PI3K-Akt and MAPK signaling pathways. Chem Biol Interact 2005;156:141-150.
51. Deng Y, Ren X, Yang L, Lin Y, Wu X. A JNK-dependent pathway is required for TNFα-induced apoptosis. Cell 2003;115:61-70.
52. Tournier C, Hess P, Yang DD, Xu J, Turner TK, Nimnual A, et al. Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. Science 2000;288:870-874.
53. Single B, Leist M, Nicotera P. Simultaneous release of adenylate kinase and cytochrome c in cell death. Cell Death Differ 1998;12:1001-1003.
54. Tsuruta F, Sunayama J, Mori Y, Hattori S, Shimizu S, Tsujimoto Y, et al. JNK promotes Bax translocation to mitochondria through phosphorylation of 14‐3‐3 proteins. EMBO J 2004;23:1889-1899.
55. Uehara T, Bennett B, Sakata ST, Satoh Y, Bilter GK, Westwick JK, et al. JNK mediates hepatic ischemia reperfusion injury. J Hepatol 2005;42:850-859.
56. Uehara T, Peng XX, Bennett B, Satoh Y, Friedman G, Currin R, et al. c-Jun N-terminal kinase mediates hepatic injury after rat liver transplantation. Transplantation 2004;78:324-332.
57. Marderstein EL, Bucher B, Guo Z, Feng X, Reid K, Geller DA. Protection of rat hepatocytes from apoptosis by inhibition of c-Jun N-terminal kinase. Surgery 2003;134:280-284.
58. Manna SK, Mukhopadhyay A, Aggarwal BB. IFN-α suppresses activation of nuclear transcription factors NF-κB and activator protein 1 and potentiates TNF-induced apoptosis. J Immunol 2000;165:4927-4934.
59. Kobayashi M, Takeyoshi I, Yoshinari D, Matsumoto K, Morishita Y. P38 mitogen-activated protein kinase inhibition attenuates ischemia-reperfusion injury of the rat liver. Surgery 2002;131:344-349.
60. Deng J, Feng J, Liu T, Lu X, Wang W, Liu N, et al. Beraprost sodium preconditioning prevents inflammation, apoptosis, and autophagy during hepatic ischemia-reperfusion injury in mice via the P38 and JNK pathways. Drug Des Devel Ther 2018;12:4067-4082.
Iranian Journal of Basic Medical Sciences
Volume 25, Issue 11
November 2022
Pages 1373-1381

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