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Knockdown of Usp13 Promotes Palmitic Acid-induced Apoptosis in Mouse Hepatocytes |
WANG Ting1,LIU Kai2,LI Ke-ying3,CHEN Xu2,REN Guang-ming2,**(),YANG Xiao-ming1,2,3,**() |
1 School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China 2 Beijing Institute of Lifeomics, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China 3 Qingdao University, Qingdao 266071, China |
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Abstract Objective: To study the effect of ubiquitin-specific protease 13 (Usp13) on the apoptosis of mouse hepatocytes stimulated by palmitic acid,and to explore the underlying molecular mechanism. Methods: Mouse primary hepatocytes were isolated by two-step collagenase perfusion technique,and then stimulated with palmitic acid after cultured in vitro.Lipid accumulation was detected by triglyceride quantitative method and oil red O staining. Cell viability was measured by MTS and LDH assay. The apoptosis of hepatocytes was tested by Annexin V-FITC/7-AAD assay. Finally the expression levels of anti-apoptotic genes and pro-apoptotic genes were detected by qPCR and the activation of Caspase-3 was detected by Western blotting. Results: Knockdown of Usp13 did not affect the accumulation of triglyceride in palmitic acid-induced hepatocytes,but the cell viability was decreased and apoptosis was increased.Studies show that knockdown of Usp13 resulted in the down-regulated expression of anti-apoptotic genes Bcl-2 and Bcl-xl, and the up-regulated expression of pro-apoptotic genes Bid and p53.The Caspase-3 activity in Usp13 knockdown hepatocytes was enhanced after being exposed to palmitic acid. Conclusion: This study revealed the mechanism of Usp13 regulating the apoptosis of hepatocytes induced by palmitic acid, which laid a foundation for further study of Usp13 regulating the occurrence and development of non-alcoholic steatohepatitis.
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Received: 08 December 2021
Published: 05 May 2022
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Corresponding Authors:
Guang-ming REN,Xiao-ming YANG
E-mail: max19920503@163.com;xiaomingyang@sina.com
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