Roles Of MiR-486 In The Proliferation And Fibrosis Of Cardiac Fibroblast

Objective To investigate roles and mechanisms of miR-486 in cardiac fibrosis.Methods(1)To establish a mouse model of cardiac remodeling following myocardial infarction.We used qPCR to detect the differential expression of miR-486 in the model.(2)We increased or inhibited the expression of miR-486 in NRCF cells,then EdU staining,to detect the influence of miR-486 to the proliferation of NRCF.(3)We first increased or inhibited the expression of miR-486 in NRCF cells.Then we used qPCR to detect the differential expression of α-sma and col-1 while used Western blot to detect the differential expression of α-sma and col-1 in protein level.(4)We increased or inhibited the expression of miR-486 in NRCF cells,and SMAD2 expression level was detected by Western blot,then co-transfected with miR-486 inhibitor and SMAD2 siRNA can rescue the promotion of NRCF proliferation,observing whether SMAD2 is a target gene of mi R-486.Results(1)miR-486 expression would be down regulated in the model of cardiac remodeling following myocardial infarction.(2)Overexpression of mi R-486 can inhibit the EdU positive rate of NRCF.Down regulation of miR-486 can increase the EdU positive rate of NRCF.(3)a.qPCR:Overexpression of miR-486 can inhibit the expression of α-sma and col-1 while down regulating miR-486 can increase the expression of them.b.Western blot: Overexpression of mi R-486 can inhibit the expression of α-sma and col-1 while they are increased when down regulating miR-486.(4)Overexpression of miR-486 can inhibit the expression of SMAD2 while it is increased when down regulating miR-486.Down regulating miR-486 can increase the EdU positive rate of NRCF,and SMAD2 si RNA can inhibit the EdU positive rate of NRCF,while co-transfected with miR-486 inhibitor and SMAD2 siRNA can rescue the promotion of NRCF proliferation.Conclusions(1)The expression level of mi R-486 was down-regulated in the model of cardiac remodeling following myocardial infarction.(2)mi R-486 can inhibit the proliferation of nrcf.(3)miR-486 can inhibit nrcf transferring into fibrosis.(4)SMAD2 was a target gene of miR-486.