Blood Circulation Has Been Effective Ingredient Fibroblast Wnt Signaling Pathway In Myocardial High Glucose

Diabetic cardiomyopathy (diabetic cardiomyopathy, DCM) is independent of the complications of diabetes and myocardial fibrosis is one of the pathogenic factor. Myocardial fibrosis is accompanied by decreased collagen network remodeling and impairment of cardiac function, than cause increased myocardial stiffeness, arrhythmia, ventricular diastolic dysfunction, deceased coronary flow reserve, even cause sudden death. High blood glucose is an important inducement of myocardial fibrosis. The doctor of traditional Chinese medicine thinks that diabetes belongs to the category of "Xiaoke", which cause is Yin deficiency and flourishing and throughout the pathogenesis of blood stasis. Supplementing qi and nourishing Yin, promoting blood circulation and removing blood stasis is the basic treatment of diabetic cardiomyopathy. Abnormal activation of Wnt pathway may be one of the causes of myocardial fibrosis. This topic discussion the anti myocardial fibrosis mechanism about the effective component of promoting blood circulation and removing blood stasis to the Wnt pathway.The first part Myocardial fibroblasts proliferation model have established by induced with high glucoseBy improving the differential adhesion method, successfully cultured neonatal rat cardiac fibroblasts. We establish the proliferation cell model successfully induced by high glucose. Then choose the optimal concentration of glucose by MTT method to the proliferation induced by high glucose. To provide the experimental basis for further research of experiment.The second part The effect on Wnt signal pathway in cardiac fibroblasts which cultured by high glucose of DHCBy high glucose in vitro fibroblast proliferation model induced by myocardial, then detection of cell viability by CCK8assay, measurement of cell proliferation by MTT assay, the cell cycle detected by flow cytometry, to observe the cell collagen secretion ability of the type Ⅰ、type III and TGF-β1, detection of the protein expression of β-catenin, GSK-3β and P-GSK-3β by western blot, to observe the effects of dehydrocorydaline on fibroblast Wnt signal pathway. The experimental results show that, DHC can inhibit cell proliferation; reduce type I, type III collagen and TGF-β1secretion; inhibit the expression of P-catenin, upregulation of the expression of P-GSK-3β. The results show that DHC has inhibitory effect on cultured myocardial cells into Wnt signal pathway.The third part The effect on Wnt signal pathway in cardiac fibroblasts which cultured by high glucose of Sa ABy high glucose in vitro fibroblast proliferation model induced by myocardial, then detection of cell viability and measurement of cell proliferation by MTT assay, the cell cycle detected by flow cytometry, to observe the cell collagen secretion ability of the type Ⅰ、type III and TGF-β1, detection of the protein expression of β-catenin, GSK-3P and P-GSK-3β by western blot, to observe the effects of Salvianolic acid A on fibroblast Wnt signal pathway. The experimental results show that, Salvianolic acid A can inhibit cell proliferation; reduce type I, type III collagen and TGF-β1secretion; inhibit the expression of β-catenin, upregulation of the expression of P-GSK-3β. The results show that Sa A has inhibitory effect on cultured myocardial cells into Wnt signal pathway.The fourth part The effect on Wnt signal pathway in cardiac fibroblasts which cultured by high glucose of Ginsenoside ReBy high glucose in vitro fibroblast proliferation model induced by myocardial, then detection of cell viability and measurement of cell proliferation by MTT assay, the cell cycle detected by flow cytometry, to observe the cell collagen secretion ability of the type Ⅰ、type Ⅲ and TGF-β1, detection of the protein expression of P-catenin, GSK-3β and P-GSK-3β by western blot, to observe the effects of Ginsenoside Re on fibroblast Wnt signal pathway. The experimental results show that, Ginsenoside Re can inhibit cell proliferation; reduce type I, type III collagen and TGF-β1secretion; inhibit the expression of β-catenin, upregulation of the expression of P-GSK-3β. The results show that Ginsenoside Re has inhibitory effect on cultured myocardial cells into Wnt signal pathway.The fifth part The effect on Wnt signal pathway in cardiac fibroblasts which cultured by high glucose of Ginsenoside RglBy high glucose in vitro fibroblast proliferation model induced by myocardial, then detection of cell viability and measurement of cell proliferation by MTT assay, the cell cycle detected by flow cytometry, to observe the cell collagen secretion ability of the type Ⅰ、type Ⅲ and TGF-β1, detection of the protein expression of β-catenin,GSK-3β and P-GSK-3β by western blot, to observe the effects of Ginsenoside Rgl on fibroblast Wnt signal pathway. The experimental results show that. Ginsenoside Rgl can inhibit cell proliferation; reduce type I, type III collagen and TGF-β1secretion; inhibit the expression of β-catenin, upregulation of the expression of P-GSK-3(3at the molecular level. The results show that Ginsenoside Rgl has inhibitory effect on cultured myocardial cells into Wnt signal pathway. The research shows that, the effective component of promoting blood circulation and removing blood stasis recipe(DHC, Salvianolic acid A, Ginsenoside Re, ginsenoside Rgl) have different degrees of inhibition on in vitro cultured cardiac fibroblasts model. Myocardial fibrosis in diabeticis closely related to abnormal activation of Wnt signaling pathway at the same time. In the process of myocardial fibrosis in diabetic, Wnt signaling pathway and TGF-β1signal pathway may have synergistic effects. These results are significance for treatment of myocardial fibrosis in diabetic.