扶正化瘀胶囊调节特异性蛋白1相关通路抗心肌纤维化作用机制研究

Research on the Mechanism of Fuzheng Huayu Capsule Regulating Specific Protein 1 Related Pathway Against Myocardial Fibrosis

目的:观察扶正化瘀胶囊对心梗大鼠特异性蛋白1相关通路的影响,探讨其防治心肌纤维化的可能作用机制。方法:结扎左冠脉前降支建立大鼠急性心梗模型,分为模型组和扶正化瘀组。假手术组只穿线不结扎。扶正化瘀组按0.4 g/(kg·d)灌胃,模型组和假手术组给予等体积去离子水,共8周。HE和Masson染色检测病理学变化;Western blot和实时荧光定量PCR法分别检测特异性蛋白1(Sp1)、Gata4、转化生长因子(TGF)-β1、Ⅰ型胶原(ColⅠ)、血小板反应蛋白1(Tsp1)及其mRNA表达。结果:与假手术组比较,模型大鼠心肌细胞破碎,纤维结缔组织大量增生,胶原容积分数(CVF)增加(P<0.01),TGF-β1、ColⅠ、Tsp1 mRNA及Sp1、Gata4、TGF-β1、ColⅠ蛋白表达明显升高(P<0.05或P<0.01),Tsp1蛋白表达降低(P<0.01);与模型组比较,扶正化瘀组细胞膜相对完整,胶原相对减少,CVF下降(P<0.01),Sp1、Gata4、TGF-β1、ColⅠ、Tsp1 mRNA及Gata4、ColⅠ蛋白表达明显降低(P<0.05或P<0.01)。结论:扶正化瘀胶囊在一定程度上改善心梗后心肌纤维化的作用机制可能与Sp1-Gata4-ColⅠ-Tsp1轴有关。

Objective:To observe the effect of Fuzheng Huayu Capsule on specific protein 1 related pathway in myocardial infarction rats,and to explore its possible mechanism of prevention and treatment of myocardial fibrosis.Methods:Rats model of acute myocardial infarction were established by ligating the left anterior descending coronary artery,which were divided into model group and Fuzheng Huayu group.The sham operation group only threaded without ligation.The Fuzheng Huayu group was given intragastrically by 0.4 g/(kg·d),the model group and the sham operation group were given equal volume of deionized water for 8 weeks.Pathological changes were detected by HE and Masson staining.Western blot and real-time fluorescent quantitative PCR were used to detect the protein and mRNA expression of specific protein 1(Sp1),Gata4,transforming growth factor(TGF)-β1,typeⅠcollagen(ColⅠ)and thrombospondin 1(Tsp1),respectively.Results:Compared with the sham operation group,the model rats had myocardial cells broken,fibrous connective tissue proliferated,and collagen volume fraction(CVF)increased(P<0.01),the expression of TGF-β1,ColⅠ,Tsp1 mRNA and the expression of Sp1,Gata4,TGF-β1,and ColⅠprotein were significantly increased(P<0.05 or P<0.01),and the expression of Tsp1 protein was decreased(P<0.05).Compared with the model group,the cells membrane of Fuzheng Huayu group were relatively intact,collagen was relatively reduced,and CVF decreased(P<0.01),the expression of Sp1,Gata4,TGF-β1,ColⅠ,Tsp1 mRNA and the expression of Gata4,ColⅠprotein were significantly decreased(P<0.05 or P<0.01).Conclusion:The mechanism of Fuzheng Huayu Capsule in improving myocardial fibrosis after myocardial infarction may be related to the Sp1-Gata4-ColⅠ-Tsp1 axis.