Study On The Effect Of Danshen Tongluo Jiedu Decoction Combined With METTL3 On Angiogenesis Of CMECs In Rats With Hypoxia/reoxygenatio

Objective: To investigate the effect and mechanism of Danshen Tongluo Jiedu Decoction combined with methyltransferase-like 3(METTL3)on angiogenesis of myocardial microvascular endothelial cells(CMECs)in rats with hypoxia/reoxygenation(H/R).Methods:1.The H/R injury CMECs model was established,and the cells were randomly divided into blank group,model group,silence group,overexpression group,Chinese medicine + silence group,Chinese medicine + overexpression group.2.LDH kit was used to detect the content of lactate dehydrogenase(LDH)in the supernatant of CMECs.3.Real-time fluorescence quantification-polymerase chain reaction(RT-PCR)was used to detect the transfection effect of METTL3 lentivirus.4.CMECs migration in each group was observed by cell scratch test.5.The proliferation ability of CMECs in each group was detected by CCK-8 method.6.m RNA expression of vascular endothelial growth factor(VEGF)in CMECs of each group was detected by real-time fluorescence quantification and polymerase chain reaction(RT-PCR).7.The protein expression of endothelial nitric oxide synthase(e NOS)in CMECs of each group was detected by Western blot.Results:1.LDH detection results showed that compared with blank group,LDH content in model group was significantly increased(P<0.01).2.The results of RT-PCR detection of METTL3 lentivirus showed that: compared with model group,m RNA expression of METTL3 in overexpression group was significantly increased(P<0.01),while m RNA expression of METTL3 in silence group was significantly decreased(P<0.01).3.Cell scratch test showed that CMECs mobility in model group was significantly decreased compared with blank group(P<0.01).Compared with model group,cell mobility in silencing group was decreased(P<0.05),while CMECs mobility in overexpression group,Chinese medicine+ overexpression group and Chinese medicine + silencing group was significantly increased(P<0.01).4.CCK-8 test results showed that the activity of CMECs in model group was significantly decreased compared with blank group(P< 0.01).Compared with the model group,the cell viability of the silencing group was decreased(P<0.05),and the CMECs activity of the overexpression group,the Chinese medicine + overexpression group and the Chinese medicine + silencing group was significantly increased(P<0.01).5.The results of RT-PCR showed that compared with blank group,the expression of VEGF in model group was significantly decreased(P< 0.01).Compared with model group,VEGF expression in overexpression group,Chinese medicine + overexpression group and Chinese medicine + silence group was significantly increased(P< 0.01,P<0.05).6.Western blot results showed that compared with blank group,e NOS protein expression in model group was significantly decreased(P<0.01).Compared with model group,the expression of silent histone decreased(P<0.05),but the expression of overexpression group,Chinese medicine +overexpression group,Chinese medicine + silent histone increased significantly(P<0.01).Conclusion:1.Hypoxia/reoxygenation model can damage CMECs.2.Danshen Tongluo Jiedu Decoction combined with METTL3 can improve the proliferation and migration ability of anoxic/reoxygenated CMECs,and has obvious protective effect on damaged CMECs.3.The protective effect of Danshen Tongluo Jiedu Decoction combined with METTL3 on hypoxic/reoxygenated CMECs may be related to up-regulation of VEGF and e NOS.