| Background:Chronic heart failure(CHF)is the final destination of all kinds of cardiovascular disease.The prognosis of CHF is poorer,at the same time the death rate of CHF is higher.CHF is called as the most important chronic cardiovascular disease in twenty-first Century.As is known to us,Ventricular remodeling is the basic mechanism of CHF,and myocardial fibrosis is the most important pathological changes in the process of ventricular remodeling.A large number of researches suggested that many factors involved in the process of myocardial fibrosis.Transforming growth factor beta 1(TGF-β1)has the most intimate relationship with myocardial fibrosis.TGF-β1 could induce the cell proliferation,collagen secretion and differentiation of cardiac fibroblasts in rats.TGF-β1 also cause the unblanance of synthesis and degradation of extracellular matrix(ECM)which leads to myocardial fibrosis in the process of ventricular remodeling.Therefore,it is the key to select a drug which could delay the development of myocardial fibrosis to prevent the process of ventricular remodeling.Jia-shen prescription(JSP),a kind of Chinese medicine,has been routinely used in treating patients with heart failure.JSP has the function of Yiqiwenyang and Huoxue lishui.Experimental studies has shown that JSP can protect rats from hypoxia and hypothermia and promote diuresis in rats.This study was aimed to determine the effects of JSP on TGF-β1 induced cell proliferation,collagen secretion and differentiation of cardiac fibroblasts in rats,At the same time,explore its possible mechanisms involved.Objective: CFs were obtained from neonatal SD rats and digested by trypsin.JSP was pre-treated the CFs with or without the 10 ng/m L TGF-β1 for 24 hours.This study was aimed to determine the effects of JSP on TGF-β1 induced cell proliferation,collagen secretion and differentiation of cardiac fibroblasts in rats.Moreover,the expressions of Smad2/3 m RNA and P-Smad2/3 protein were also analyzed.At the same time,explore its possible mechanisms involved.Methods : 1.Isolation and culture of cardiac fibroblasts 2.Morphological observation and identification of CFs 3.Experimental groups and intervention Screening of different concentration JSP groups:(1)0mg/ml JSP groups;(2)0.25mg/ml JSP groups;(3)0.5mg/ml JSP groups;(4)1.0mg/ml JSP groups.Experimental grouping:(1)normal group(normal culture);(2)model group(10 ng/m LTGF-β1);(3)JSP intervene group(10ng/m LTGF-β1+0.25mg/ml JSP)4.Cell Proliferation Assay 5.Immunofluorescence Assay for α-Smooth Muscle Actin and Collagen I 6.Measurement of Hydroxyproline 7.Total RNA extraction and RT-PCR for Smad2/3 8.Western Blott analyses for TGF-β1 and p-Smad2/3Results: 1.Characterization of cultured neonatal rat CFs.2.Identification of cardiac fibroblasts.3.Experimental grouping.4.Proliferation was detected by CCK-8.5.The expression of α-SMA and Collagen I of CFs was detected by immunofluorescence quantification.6.Hydroxyproline secretion of CFs was detected by Kit.7.The expression of Smad2/3 m RNA was detected by RT-PCR.8.The expression of P-Smad2/3 protein was detected by Western-blot.Conclusion: 1.On the basis of culture myocardial fibroblast,cell proliferation,collagen secretion and differentiation of cardiac fibroblasts in rats induced by 10ng/ml TGF-β1.2.On the basis of culture myocardial fibroblast,Jia-Shen Prescription inhibited cell proliferation,collagen secretion and differentiation of cardiac fibroblasts in rats induced by TGF-β1;At the same time,Jia-Shen Prescription reduced the expression level of Smad2/3 m RNA and P-Smad2/3 protein of cardiac fibroblasts in rats induced by TGF-β1.3.Jia-Shen Prescription inhibited cell proliferation,collagen secretion and differentiation of cardiac fibroblasts in rats induced by TGF-β1.So we guessed the mechanism of Jia-Shen Prescription depent on inhibition of TGF-β1/Smads signalling way. |
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