Effects Of Intermedin_1-53 Induced By AngⅡ On Myocardial Fibrosis And Extracellular Matrix

Objective:Myocardial fibrosis is lead to cardiac insufficiency, arrhythmia, and chronic heart failure is difficult to reverse one of the major pathological damage.Heart failure when damaged cardiac fibroblasts collagen synthesis and degradation of the balance between, changed the myocardial extracellular matrix structure and composition are important cause of ventricular remodeling.This experiment adopts the AngII induced myocardial fibroblasts(fibroblasts, CFS), fibrosis model was established, through the intervention Intermedin1-53 to observe Intermedin1-53 AnglI induced myocardial fibroblasts(CFS) collagen synthesis and the effect of extracellular matrix structure and composition, argument IMD1-53 is important for the body anti fibrosis factor, to delay the ventricular remodeling, inhibiting myocardial fibrosis provide theoretical basis for the prevention and control. Methods: 1. The isolation and culture of myocardial fibroblasts: clean level 1-4 days were male SD rats, low-temperature anesthesia, remove the ventricular muscle tissue cut into small pieces, with pancreatic enzyme digestion method through poor cultivation wall in cardiac fibroblasts. 2.Grouping: take the original generation of cardiac fibroblasts culture, good condition, wait 24 hours later, with CFS AngII stimulation. The experiment is divided into five groups:(1) normal control group(normal control) : 10% fetal bovine serum DMEM culture;(2) AngII function groups: DMEM medium, 10% fetal bovine serum to join concentration for 10-6mmol/L Ang IIgroup;(3) AngII + IMD1- low concentration groups: 53 DMEM medium, 10% fetal bovine serum to join concentration for 10-6 mmo l/l Ang IIgroup, 10 to 9 tendency at the same time to join/l IMD1-53;(4) AngII + IMD1-53 concentration groups: DMEM medium, 10% fetal bovine serum to join concentration for 10-6 mmo l/l Ang IIgroup, 10-8 tendency at the same time to join/l IMD1-53;(5) AngII + IMD1-53 high concentration groups: DMEM medium, 10% fetal bovine serum to join concentration for 10-6 mmo l/l Ang IIgroup. 3.Western blot imprinting method to detect myocardial fibroblasts Ⅰ and Ⅲ type collagen MRNA and protein expression and matrix components such as fibrils protein- 2 MRNA and protein expression of binding protein and fiber. 4. SYBR Green Ⅰ real-time fluorescent quantitative PCR method(real time PCR) to detect IMD1-53 receptor like receptor(CRLR) and transforming growth factor beta(TGF- beta) mRNA expression changes. Results: 1. IMD1-53 of AngII under the intervention of cardiac fibroblasts in type 1, type III collagen, collagen fibrils protein- 2 and fiber combined with protein content, the influence of the test shows that: compared with control group, group AngII myocardial fibroblasts under the action of AngII type I collagen, type III collagen, protein fiber binding protein and fibrils- 2 levels were significantly higher than the normal control group(P < o. 01); AngII + IMD1-53, concentration of 10 to 9 in the tendency for L type I collagen, type III collagen, protein fiber binding protein and fibrils- 2 levels were significantly lower than the AngII group(P < o. 01), while no statistical differences compared with normal control group(P > 0.05), and with the IMD1-53, concentration increases, the performance of concentration dependence to inhibit AngII induced myocardial fibroblasts collagen synthes. 2. As IMD1-53 concentration, the greater the inhibition of the expression of fibroblast TGF- beta, the more obvious, a dose dependent(P < 0.05). 3.Real-time fluorescent quantitative PCR results showed that in the control group as a reference, IMD role experimental ventricular muscle CRLR mRNA levels were significantly higher(n = 3, P < 0.05 or P < 0.05), Conclusion:1. IMD1-53 by reducing the type I collagen, type III collagen, such as myocardial fibroblasts composition and fibrinogen and extracellular matrix components, such as fiber combined with protein content, may reach the role of inhibition of myocardial fibrosis, and inhibition of concentration dependence IMD1-53 2.IMD1-53 promote fibrosis factor can be adjusted by the expression of TGF- beta, may inhibit Ang Ⅱ induced myocardial fibroblasts collagen synthesis and secretion of extracellular matrix, thereby IMD1-53 against myocardial fibrosis has certain intervention effect.