Experimental Studies Of RNF122 In Myocardial Ischemia And Reperfusion Injury

Objective Oxygen-reoxygenation treatment was used to simulate the pathological process of diabetes mellitus,myocardial ischemia-reperfusion,and diabetes mellitus combined with myocardial ischemia-reperfusion.The m RNA expression level of RNF122 in each group was detected,and the specific siRNA of RNF122 was used to down-regulate the expression in each model.The relevant indicators were detected in the group to understand the effects of RNF122 on cardiomyocyte apoptosis,oxidative stress,and ERK pathway.Methods(1)Selection of rat cardiomyocyte cell line H9c2 The model of high glucose,hypoxia and reoxygenation,and high glucose combined with hypoxia and reoxygenation were established to simulate the pathological process of diabetes,myocardial ischemia and reperfusion,and diabetes combined with myocardial ischemia and reperfusion,and the cell growth of each model was detected by flow cytometry.(2)Using real-time quantitative PCR to detect changes in m RNA expression of RNF122 in different cell models;(3)Specific siRNA transfected with RNF122 down-regulates its expression.Western blot and PCR techniques are used to detect the oxidation of cells after down-regulation.Intensity,apoptosis,and changes in ERK pathway-related indicators.Results(1)High-glucose,hypoxia-reoxygenation and high-glucose combined hypoxia-reoxygenation models were successfully established.Flow cytometry showed that apoptosis increased with prolonged hypoxia,but cell death rate There was no significant change,suggesting that cell hypoxia mainly leads to apoptosis rather than necrosis;6 hours after hypoxia,the cells were reoxygenated,and as the time of reoxygenation prolonged,apoptosis was further aggravated,indicating that reoxygenation aggravated the deficiency.(2)RT-PCR was used to detect the change of m RNA expression of RNF122 in each cell model,suggesting that high glucose(HG)and hypoxia-reoxygenation(H/R)treatment resulted in intracellular RNF122 m RNA level.Significantly up-regulated,and high glucose combined with hypoxia and reoxygenation will further increase its expression;(3)Detecting the expression of oxidative stress-related factor GRP78 by down-regulating the expression of RNF122,suggesting that downregulation of RNF122 can significantly improve cell oxidation.Excited situation(4)The expression of apoptosis-related factor Cleaved-casp3 was detected by down-regulating the expression of RNF122,suggesting that the downregulation of RNF122 could significantly improve the apoptosis of cells;(5)To detect the ERK pathway in cells after down-regulating the expression of RNF122;The protein expression changes of related markers(ERK1,p-ERK1,ERK2,p-ERK2)showed that the protein level of ERK1/2 did not change significantly after down-regulation of the RNF122 gene level,but the protein expression of p-ERK1/2 was significantly up-regulated.Conclusion(1)RNF122 causes apoptosis and oxidative stress in the pathology of diabetes mellitus,myocardial ischemia-reperfusion injury,and diabetes combined with myocardial ischemia-reperfusion injury;(2)RNF122 regulates the pathological process of diabetes mellitus,myocardial ischemia-reperfusion injury,and myocardial ischemiareperfusion injury in diabetes by regulating the phosphorylation of the ERK pathway.