The Role Of MiR-26a In Regulating The Expression Of FLI1 In The Treatment Of Fibrosis Of Scleroderma By Bushenyijing Therapy

ObjectiveIn order to elucidate the mechanism of Bushen Yijing therapy in treating scleroderma for its clinical application.Furthermore,for the sake of provided a strong theoretical basis for promoting the inheritance and innovation of excellent TCM treatment.The research studied the effect of Bushenyijing therapy on the expression of FLI1 regulated by miR-26 a and epigenetic mechanism of scleroderma.What’s more,The study provided the new ideas and new ways for the modern treatment of scleroderma through studying the role of miR-26 a in the mechanism of fibrosis of scleroderma and exploring new intervention targets for the treatment of scleroderma.MethodsThe fibroblasts were taken from patients with systemic scleroderma(SFB)and the normal human skin fibroblasts were isolated from healthy people(NFB).MiR-26 a mimics and MiR-26 a inhibitor were used to regulate the expression of miR-26 a of fibroblasts.Then the expression of miR-26 a,FLI1,Collagen I and Fibronectin in SFB group were detected before and after transfection.Hereafter,all fibroblasts were divided into 6 groups,including NFB group,which were supplied with serum.SFB were divided into 5 groups,which were added with serum,astragaloside IV,containing serum of Ruanpi Decoction,containing serum of Ruanpi Decoction + miR-26 a mimics,containing serum of Ruanpi Decoction + miR-26 a mimics inhibitor,respectively.At last,the expression of miR-26 a,FLI1,Collagen I and Fibronectin in each group was detected.Results1.It successfully established scleroderma and normal human dermal fibroblast cell lines and they were identified as fibroblasts by immunocytochemistry: CK(-),Vimentin(+).2.Compared with NFB,the overexpression of miR-26 a and the low expression of FLI1 in SFB were observed,the difference was statistically significant(P<0.05).3.Compared with SFB treated with serum,miR-26 a overexpression,FLI1 low expression,Collagen I and Fibronectin expression elevationin of SFB were obtained after being transfected with miR-26 a mimics.While miR-26 a low expression,FLI1 high expression,Collagen I,Fibronectin expression decline of SFB were obtained after being transfected with miR-26 a inhibitor.The difference was statistically significant(P<0.05).4.Compared with NFB,the expression of FLI1 in SFB decreased and the expression of Collagen Iand Fibronectin increased.The expression of FLI1 aslo increased and Collagen Iand Fibronectin decreased after SFB cells were treated with drugs(astragaloside IV and containing serum of Ruanpi Decoction).After being treated with the serum of Ruanpi Decoction and miR-26 a inhibitor,the expression of FLI1 was up-regulated and the expression of Collagen Iand Fibronectin was down-regulated compared with the SFB group.After being transfected with the serum of Ruanpi Decoction and miR-26 a mimics,the expression of FLI1 was not significantly different,and the expression of Collagen I and Fibronectin was higher than the SFB group.ConclusionIn the fibrolasts of sclerodema,the over-expression of miR-26 a may promote fibrosis by down-regulating the expression of FLI1.Bushenyijing therapy may play an important role in inhibiting fibrosis by down-regulating the expression of miR-26 a and up-regulating the expression of FLI1.