Study On The Effect And Mechanism Of Danzhi Jiangtang Capsule In Interfering With High Glucose-Induced Myocardial Fibrosis

ObjectiveTo study the effects of Danzhi Jiangtang Capsule on the myocardial tissue fibrosis and the regulation of high glucose-induced myocardial fibroblast proliferation in diabetic model rats,and to explore its mechanism through the TLR4-My D88-NF-?B signaling pathway.MethodsIn vivo experiment:Clean-type SD rats were given high-fat diets combined with streptozotocin to prepare a type 2 diabetic rat model.The rats successfully modeled were randomly divided into model groups?equivalent volume of distilled water?.The high-dose group?270,540,1080 mg/kg?,the positive drug group?150 mg/kg,metformin?,and a control group with ordinary feed?equivalent volume of distilled water?,drug intervention for 8 weeks,the heart was dissected,HE and Masson staining,Collagen I and Collagen?immunohistochemical detection were used to observe the morphological changes of myocardial tissue in each group,and to study the effect of Danzhijiangtang capsule on myocardial fibrosis in diabetic model rats.In vitro experiments:Neonatal SD neonatal rat myocardial fibroblasts were isolated and cultured to establish a high glucose-induced myocardial fibroblast proliferation model.After the intervention of Danzhi Jiangtang Capsule-containing serum and TLR4 specific blocker,cell viability and cell scratches were observed by MTT experiments.Experiments and immunofluorescence were used to observe cell proliferation.Western blot and RT-q PCR were used to detect changes in TLR4-My D88-NF-?B signaling pathway and myocardial fibrosis-related proteins and genes,and to clarify that TLR4-My D88-NF-?B signaling pathway regulates high glucose induction The molecular mechanism of myocardial fibroblast proliferation reveals the effect of Danzhijiangtang capsule on high glucose-induced myocardial fibrosis and its relationship with TLR4-My D88-NF-?B signaling pathway.ResultsThe first part of the effect of Danzhi Jiangtang capsule on myocardial injury in diabetic ratsMorphological observation of myocardial tissue:Compared with the control group,the results of HE and Masson showed that myocardial rupture,increased fibroblasts with inflammatory infiltration,collagen deposition with interstitial fibrosis,and immunohistochemistry in the model group.A large amount of Collagen I and Collagen III deposits can be seen in the myocardial tissue of model rats;compared with the model group,the myocardial tissues of the myocardial tissues of the Danzhijiangtang capsule group and the myocardial tissues of the positive drug group showed HE and Masson's results.The directions tend to be the same,the myocardial fiber breakage is reduced,the cells are arranged neatly,the collagen proliferation is significantly reduced,and there is a small amount of inflammatory cell infiltration.Immunohistochemical staining shows that the expression of Collagen I and Collagen?is significantly reduced.The second part of the mechanism of Danzhi Jiangtang Capsules regulating TLR4-My D88-NF-?B signaling pathway against high glucose-induced proliferation of cardiac fibroblasts?1?Different glucose concentrations?5.5,15,30,45,60 mmol/L?and hypertonic group?mannitol?intervention for 48 h.Observation of OD values of myocardial fibroblasts showed that they were similar to the normal group?5.5mmol/L?.Compared with the high glucose model group?30mmol/L?,the OD value increased significantly after 48h intervention?P<0.01?.The proliferation model of myocardial fibroblasts induced by high glucose was successfully prepared;Danzhi Jiangtang Capsules with different concentrations of serum(5%?10%,15%,20%,25%?intervention for 48 hours,only25%concentration group showed cytotoxicity;?2?Compared with the control group,the migration inhibition rate of myocardial fibroblasts in the high glucose model group was significantly reduced?P<0.05?,TGF-?₁,Vimentin,Collagen I and Collagen III protein expression,protein and gene expression of TLR4,My D88,NF-?B p65 all significantly increased?P<0.05?;?3?Compared with the high glucose model group,TLR4 specific resistance In the intervention group of TAK-242,the proliferation of myocardial fibroblasts was reduced,and the cell migration inhibition rate was significantly increased?P<0.05?.TGF-?1,Vimentin,Collagen I,and Collagen III proteins were expressed,and TLR4,My D88,and NF-?B p65 proteins and Gene expression decreased significantly?P<0.05?;?4?Compared with the high-glucose model group,the proliferation of myocardial fibroblasts and the cell migration inhibition rate of the Danzhi Jiangtang capsule-containing serum group were significantly reduced?P<0.05?.TGF-?₁,Vimentin,Collagen I,and Collagen III protein expression,protein and gene expression of TLR4,My D88,NF-?B p65 were significantly reduced?P<0.05?.ConclusionDanzhi Jiangtang Capsule can regulate the TLR4-My D88-NF-?B signal pathway,inhibit the proliferation of myocardial fibroblasts induced by high glucose,resist myocardial fibrosis,improve the morphology of myocardial tissue in diabetic model rats,and effectively interfere with myocardium complicated by diabetes Tissue damage has a significant role in preventing and treating diabetic cardiomyopathy.