N-acetylcysteine protects septic acute kidney injury by inhibiting SIRT3-mediated mitochondrial dysfunction and apoptosis

Document Type : Original Article

Authors

Department of Intensive Care Unit, The First Affiliated Hospital of Ningbo University, Ningbo, Zhejiang Province, P.R China

10.22038/ijbms.2024.72882.15853

Abstract

Objective(s): To investigate the protective effect of N-acetylcysteine (NAC) on septic acute kidney injury (SAKI) via regulating Sirtuin3 (SIRT3)-mediated mitochondrial dysfunction and apoptosis.
Materials and Methods: By constructing SIRT3 knockout mice and culturing kidney tubular epithelial cells (KTECs), we assessed the changes of renal function and detected the protein expression of adenine nucleotide translocator (ANT), cyclophilin (CypD) and voltage-dependent anion channel (VDAC) using western-blotting, and simultaneously detected toll-like receptor 4 (TLR4), inhibitor of kappa B kinase (IKKβ), inhibitor of Kappa Bα (IκBα), and p65 protein expression. We observed mitochondrial damage of KTECs using a transmission electron microscope and assessed apoptosis by TdT-mediated dUTP Nick-End Labeling and flow cytometry. 
Results: SIRT3 deficiency led to the deterioration of renal function, and caused a significant increase in inducible nitric oxide synthase production, a decrease in mitochondrial volume, up-regulation of TLR4, IκBα, IKKβ, and p65 proteins, and up-regulation of ANT, CypD and VDAC proteins. However, NAC significantly improved renal function and down-regulated the expression of TLR4, IκBα, IKKβ, and p65 proteins. Furthermore, SIRT3 deficiency led to a significant increase in KTEC apoptosis, while NAC up-regulated the expression of SIRT3 and inhibited apoptosis.
Conclusion: NAC has a significant protective effect on SAKI by inhibiting SIRT3-mediated mitochondrial dysfunction and apoptosis of KTECs. 

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References

1. Ronco C, Bellomo R, Kellum JA. Acute kidney injury. Lancet 2019;394:1949-1964.
2. Fan H, Le JW, Sun M, Zhu JH. Sirtuin 3 deficiency promotes acute kidney injury induced by sepsis via mitochondrial dysfunction and apoptosis. Iran J Basic Med Sci 2021;24:675-681.
3. Jacob J, Dannenhoffer J, Rutter A. Acute kidney injury. Prim Care 2020;47:571-584. 
4. Fan H, Le JW, Zhu JH. Protective Effect of N-acetylcysteine pretreatment on acute kidney injury in septic rats. J Surg Res 2020; 254:125-134.
5. Bellomo R, Kellum JA, Ronco C, Wald R, Martensson J, Maiden M, et al. Acute kidney injury in sepsis. Intensive Care Med 2017; 43:816-828.
6. Zhao WY, Zhang L, Sui MX, Zhu YH, Zeng L. Protective effects of sirtuin 3 in a murine model of sepsis-induced acute kidney injury. Sci Rep 2016; 6:33201-33211.
7. Klimova N, Fearnow A, Long A, Kristian T. NAD+precursor modulates post-ischemic mitochondrial fragmentation and reactive oxygen species generation via SIRT3 dependent mechanisms. Exp Neurol 2020; 325:113144-113180.
8. Wang Z, Sun R, Wang G, Chen Z, Li Y, Zhao Y, et al. SIRT3-mediated deacetylation of PRDX3 alleviates mitochondrial oxidative damage and apoptosis induced by intestinal ischemia/reperfusion injury. Redox Biol 2020; 28:101343-101355.
9. Wei S, Gao Y, Dai X, Fu W, Cai S, Fang H, et al. SIRT1-mediated HMGB1 deacetylation suppresses sepsis-associated acute kidney injury. Am J Physiol Renal Physiol 2019; 316:F20-F31.
10. Fan H, Zhao Y, Zhu JH. S-nitrosoglutathione protects lipopolysaccharide-induced acute kidney injury by inhibiting toll-like receptor 4-nuclear factor-κB signal pathway. J Pharm Pharmacol 2019; 71:1255-1261.
11. Zhao W, Zhang L, Chen R, Lu H, Sui M, Zhu Y, et al. SIRT3 protects against acute kidney injury via AMPK/mTOR-regulated autophagy. Front Physiol 2018; 9:1526-1535.
12. Perico L, Morigi M, Benigni A. Mitochondrial Sirtuin 3 and Renal Diseases. Nephron 2016; 134:14-19.
13. Perico L, Remuzzi G, Benigni A. Sirtuin 3 in acute kidney injury. Oncotarget 2015;6:16814-16815.
14. Li Y, Ye Z, Lai W, Rao J, Huang W, Zhang X, et al. Activation of sirtuin 3 by silybin attenuates mitochondrial dysfunction in cisplatin-induced acute kidney injury. Front Pharmacol 2017;8:178-189.
15. Xu S, Gao Y, Zhang Q, Wei S, Chen Z, Dai X, et al. SIRT1/3 activation by resveratrol attenuates acute kidney injury in a septic rat model. Oxid Med Cell Longev 2016; 2016:7296092.
16. Peerapanyasut W, Kobroob A, Palee S, Chattipakorn N, Wongmekiat O. N-acetylcysteine attenuates the increasing severity of distant organ liver dysfunction after acute kidney injury in rats exposed to bisphenol A. Antioxidants (Basel) 2019;8:497-511.
17. Allison SJ. Acute kidney injury: Sirtuin 3-a master regulator of mitochondrial integrity in AKI. Nat Rev Nephrol 2015;11:197. 
18. Huang YT, Chen YY, Lai YH, Cheng CC, Lin TC, Su YS, et al. Resveratrol alleviates the cytotoxicity induced by the radiocontrast agent, ioxitalamate, by reducing the production of reactive oxygen species in HK-2 human renal proximal tubule epithelial cells in vitro. Int J Mol Med 2016;37:83-91.
19. Morigi M, Perico L, Benigni A. Sirtuins in renal health and disease. J Am Soc Nephrol 2018; 29:1799-1809.
20. Morigi M, Perico L, Rota C, Longaretti L, Conti S, Rottoli D, et al. Sirtuin 3-dependent mitochondrial dynamic improvements protect against acute kidney injury. J Clin Invest 2015;125:715-726.
21. Ouyang J, Zeng Z, Fang H, Li F, Zhang X, Tan W. SIRT3 Inactivation promotes acute kidney injury through elevated acetylation of SOD2 and p53. J Surg Res 2019;233:221-230.
22. Yuan Y, Zhu L, Li L, Liu J, Chen Y, Cheng J, et al. S-sulfhydration of sirt3 by hydrogen sulfide attenuates mitochondrial dysfunction in cisplatin-induced acute kidney injury. Antioxid Redox Signal 2019;31:1302-1319.
23. Wang Q, Xu J, Li X, Liu Z, Han Y, Xu X, et al. Sirt3 modulate renal ischemia-reperfusion injury through enhancing mitochondrial fusion and activating the ERK-OPA1 signaling pathway. J Cell Physiol 2019;234:23495-23506.
24. Li M, Li CM, Ye ZC, Huang J, Li Y, Lai W, et al. Sirt3 modulates fatty acid oxidation and attenuates cisplatin-induced AKI in mice. J Cell Mol Med 2020;24:5109-5121.
25. Huang Z, Li Q, Yuan Y, Zhang C, Wu L, Liu X, et al. Renalase attenuates mitochondrial fission in cisplatin-induced acute kidney injury via modulating sirtuin-3. Life Sci 2019;222:78-87.
26. Zhang Q, Liu X, Li N, Zhang J, Yang J, Bu P. Sirtuin 3 deficiency aggravates contrast-induced acute kidney injury. J Transl Med 2018;16:313-324.
27. Tan C, Gu J, Li T, Chen H, Liu K, Liu M, et al. Inhibition of aerobic glycolysis alleviates sepsis-induced acute kidney injury by promoting lactate/Sirtuin 3/AMPK-regulated autophagy. Int J Mol Med 2021; 47:19-30.
 
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 05 March 2024

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