Study on Mechanism of Cinnamomi Oleum Anti-arrhythmia of H9c2 Cells by Regulating miR-328 Gene
Objective:To investigate the possible mechanism of Cinnamomi Oleum anti-arrhythmia of H9c2 cells by regulating miR-328 gene.Methods;The miR-328 over-expression H9c2 cell line was constructed by lentivirus transfection,and the arrhythmia model of H9c2 cells and miR-328 over-expression H9c2 cells was constructed by biological function experiment system.H9c2 cells were divided into normal control group,model group,Cinnamomi Oleum group,miR-328 over-expression group and miR-328 over-expression with Cinnamomi Oleum group.After the culture was finished according to the group conditions,the relative expression of miR-328 in H9c2 cells was detected by qPCR.The contents of IL-1β,IL-6 and TGF-β were detected by ELISA.SOD activity and MDA concent were detected by colorimetry.DCFH fluorescence was used to detect the level of ROS,the apoptosis rate was detected by flow cytometry.Western Blot was used to detect the expression of L-type calcium channel-related proteins in H9c2 cells.Results:The arrhythmia model of H9c2 cells and miR-328 over-expression H9c2 cells was successfully constructed.Compared with the normal control group,the TGF-β,SOD activity and CaM,CaMK Ⅱ,p-CaMK Ⅱ protein expressions in the model group and miR-328 over-expression group were signifi-cantly decreased,the apoptosis rate,protein expressions of miR-328 and CaVl.2 and the levels of IL-1β,IL-6,MDA and ROS were sig-nificantly increased(P＜0.01).After Cinnamomi Oleum intervention,the morphology of cells in arrhythmia model group was improved,and the above indexes were significantly reversed(P＜0.01).Conclusion;Cinnamomi Oleum can improve arrhythmia H9c2 cells,and its mechanism may be related to the regulation of miR-328 gene to reduce the level of inflammatory factors,regulate oxidative stress,inhibit apoptosis and regulate L-type calcium channel protein expressions.
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