Role of GSK-3β in lipid emulsion-induced inhibition of bupivacaine-induced apoptosis in cardiomyocytes of rats: evaluation using RNA interference adenovirus infection method / 中华麻醉学杂志

ABSTRACT

Objective To evaluate the role of glycogen synthase kinase 3 beta (GSK-3β) in lipid emulsion-induced inhibition of bupivacaine-induced apoptosis in cardiomyocytes of rats using RNA interference (RNAi) adenovirus infection method.Methods H9C2 cells were transferred into 96-well cell plates at a density of 1× 105 cells/ml after culture and then divided into 8 groups (n =10 each) using a random number tablecontrol group (group C),bupivacaine group (group B),lipid emulsion group (group LE),bupivacaine plus lipid emulsion group (group B+LE),control plus GSK-3βRNAi adenovirus (GSK-3βi) group (group C+GSK-3βi),bupivacaine plus GSK-3βi group (group B+GSK-3βi),lipid emulsion plus GSK-3βi group (group LE+GSK-3βi) and bupivacaine plus lipid emulsion plus GSK-3βi group (group B+LE+GSK-3βi).ln B,LE and B+LE groups,the cells were incubated with culture medium containing 1 mmol/L bupivacaine,1％ lipid emulsion and 1 mmol/L bupivacaine plus 1％ lipid emulsion,respectively.In C+GSK-3βi,B+GSK-3βi,LE+GSK-3βi and B+LE+GSK-3βi groups,the cells were incubated with the drugs mentioned above on 2nd day after being infected by adenovirus.At 24 h after incubation with drugs,the expression of Bax and Bcl-2 was determined by Western blot,and the apoptosis rate was calculated using DAPI staining.Results Compared with group C,the expression of Bax was significantly upregulated,the expression of Bcl-2 was down-regulated,and the apoptosis rate was increased in group B (P＜0.05).Compared with group B,the expression of Bax was significantly down-regulated,the expression of Bcl-2 was up-regulated,and the apoptosis rate was decreased in group B+LE (P＜0.05).Compared with group B+LE,the expression of Bax was significantly up-regulated,the expression of Bcl-2 was downregulated and the apoptosis rate was increased in group B+LE+GSK-3βi (P＜0.05).Conclusion The mechanism by which lipid emulsion inhibits bupivacaine-induced apoptosis in cardiomyocytes of rats is associated with GSK-33.