Effect of bupivacaine mediated apelin on gene expression of rat cardiomyocytes
Objective To investigate the effect of bupivacaine on Apelin/APJ pathway and subsequent gene expression in Sprague-Dawley(SD)rat cardiomyocytes.Methods H9c2 cardiomyocytes were cultured in vitro and treated with 0-1 mmol/L bupivacaine for 6 h.The concentration of Apelin was detected by enzyme-linked immunosorbent assay(ELISA),and the expression of APJ was detected by Western blot;Fifteen adult male SD rats were randomly divided into sham group(sham group),bupivacaine 30 mg/kg group(Bupi group),bupivacaine 30 mg/kg+[Pyr1]apelin-13 0.15 mg/kg(Bupi-A)group.After corresponding treatment,myocardial tissue was taken to detect the expression of Apelin and APJ,and high-throughput sequencing was performed.Results Western blot and EUSA results showed that bupivacaine down regulated the expression of Apelin and APJ in H9c2 cardiomyocytes(all P＜0.05).The expression levels of Apelin and APJ in the myocardial tissue of SD rats in Bupi group were lower than those in sham group(all P＜0.05).The expression of Apelin in myocardial tissue of Bupi-A group was higher than that of Bupi group(P=0.006),but the expression of APJ had no significant difference(P＞0.05).High throughput sequencing revealed that compared with sham group,the top five differentially expressed genes that were significantly upregulated in Bupi group but significantly downregulated in Bupi-A group were ubiquinone NADH dehydrogenase Fe-S protein 3,enolase 1,aquaporin 1,ATP synthase F0 complex C subunit 1 lipid binding protein,and peroxidase 2(all P＜0.001);The top five genes that were significantly down regulated in Bupi group but significantly up-regulated in Bupi-A group were mesothelin,Rho GTPase activating protein 29,sten20 like kinase,carbonic anhydrase,and paraplatelet lysin(all P＜0.001).These genes were associated with increased cardiomyocyte apoptosis,energy metabolism disorders and exercise attenuation.Conclusions bupivacaine can reduce the expression of Apelin,leading to the changes of gene expression related to increased apoptosis,energy metabolism disorder and exercise attenuation in rat cardiomyocytes.
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