Abstract
The purpose of the present study is to define the role of sevoflurane (SEV) in hepatic ischemia-reperfusion (I/R) injury as well as its underlying mechanism. Initially, hepatic I/R animal models and I/R hepatocyte models were established in C57BL/6 mice and normal mouse hepatocytes (BNL CL.2) after SEV preconditioning, respectively, followed by detection of microRNA-124-3p (miR-124-3p), TRAF3, and CREB expression by RT-qPCR and Western blot analysis. In addition, miR-124-3p, TRAF3 and CREB expression in hepatocytes was altered to identify their roles in modulating the levels of glutathione transferase (GST), aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and inflammation-related factors and hepatocyte apoptosis by ELISA and flow cytometry respectively. The effects of SEV on the miR-124-3p/TRAF3/CREB axis were also verified in vitro and in vivo. IP assay was performed to verify the effect of TRAF3 on CREB ubiquitination in BNL CL.2 cells, and the cycloheximide (CHX) intervention experiment to detect the stability of CREB protein. SEV augmented the miR-124-3p expression in I/R animal and cell models. Moreover, SEV was observed to suppress I/R-induced liver damage, GST, ALT, and AST levels, hepatocyte apoptosis and inflammation. Overexpression of miR-124-3p resulted in alleviation of hepatic I/R injury, which was countered by TRAF3 overexpression. miR-124-3p targeted TRAF3, while TRAF3 promoted CREB ubiquitination and reduced protein stability of CREB. SEV could impede I/R-induced liver damage, GST, ALT, and AST levels, hepatocyte apoptosis and inflammation via mediation of the miR-124-3p/TRAF3/CREB axis in vitro and in vivo. Collectively, SEV may upregulate miR-124-3p to inhibit TRAF3 expression, thereby reducing the ubiquitination and degradation of CREB, alleviating hepatic I/R injury.
Highlights
Hepatic ischemia-reperfusion (I/R) injury is a common occurrence following blood supply restoration after blood vessel occlusion during the course of countless liver surgeries [1]
Enzyme-linked immunosorbent assay (ELISA) depicted that ALT, AST, and GST levels were increased by SEV + miR-124-3p inhibitor in H2O2-treated hepatocytes, whereas oe-CREB declined the said levels in H2O2-treated hepatocytes in the presence of SEV + miR-124-3p inhibitor
Flow cytometric data demonstrated that hepatocyte apoptosis was notably increased upon SEV + miR-124-3p inhibitor in H2O2-treated hepatocyte, which was reversed by SEV + miR-1243p inhibitor + oe-CREB (Fig. 5C)
Summary
Hepatic ischemia-reperfusion (I/R) injury is a common occurrence following blood supply restoration after blood vessel occlusion during the course of countless liver surgeries [1]. Hepatic I/R injury is recognized as the leading cause of abnormal liver function and hepatic failure, whereas there are only a limited number of options to effectively tackle this major complication of liver surgery [2]. Hepatic I/R injury has long been a major cause of liver dysfunction and failure after surgical procedures [3, 4]. Hepatic I/R injury implicates a persistent pro-oxidant and pro-inflammatory mechanisms that result in hepatocyte dysfunction and inflammatory or apoptotic responses [5, 6]. Pretreatment with sevoflurane (SEV) has been reported to protect liver tissues against I/R injury, as well as confer protection to hepatocytes against I/R-triggered necrosis [7]. The molecular mechanisms of SEV underlying the highly-beneficial hepatoprotection remain largely unknown
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