冠心舒通胶囊抑制TGF-β/Smad信号通路改善异丙肾上腺素诱导的大鼠心脏重塑机制研究

Guanxin Shutong Capsule Improves Isoproterenol-induced Cardic Remodeling by Repressing the TGF-β/Smad Signaling Pathway

目的探讨冠心舒通胶囊对异丙肾上腺素(isproterenol,ISO)诱导的大鼠心脏重塑的改善作用和对TGF-β/Smad信号通路的影响。方法将SD雄性大鼠随机分为6组,即正常对照组,模型组,冠心舒通胶囊低、中、高剂量组(1.2、2.4、4.8g·kg^-1),普萘洛尔组(0.5g·kg^-1),采用大鼠皮下多点注射ISO(5mg·kg^-1)连续7d建立心脏重塑模型,造模启动后第2天开始灌胃给药,每日1次,连续1个月,采用心脏多普勒超声检测大鼠左室射血分数(EF)、左室短轴缩短率(FS)、左室舒张末期容积(EDV)、左室收缩末期容积(ESV)、左室舒张末期直径(LVIDd)、左室后壁厚度(LVPWd),测定全心质量指数(HMI),左心室质量指数(LVMI),评价大鼠心脏功能;Masson染色观察大鼠心肌病理形态学变化,计算心肌胶原容积分数(CVF);生化法检测心肌组织羟脯氨酸(Hydro)的含量;酶联免疫吸附试验(ELISA)法检测血清中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的水平;蛋白免疫印迹(Western Blot)法检测心肌组织中TGF-β、Smad2/3、Caspase-3蛋白表达水平。结果与正常对照组相比,模型组大鼠心脏EF、FS降低,EDV、ESV、LVIDd、LVPWd和HMI、LVM1值均显著升高;心肌纤维化加重,CVF值及Hydro的含量升高;血清炎症因子TNF-α、IL-6水平明显增加;心肌组织中TGF-β、Smad2/3、Caspase-3蛋白表达水平升高,差异均有统计学意义(P<0.01)。与模型组相比,冠心舒通胶囊中、高剂量组可改善ISO诱导的大鼠心脏功能异常;并降低心肌纤维化水平及Hydro含量;降低血清炎症因子TNF-α,IL-6含量;同时抑制心肌组织TGF-β、Smad2/3、Caspase-3蛋白表达,差异均有统计学意义(P<0.05,P<0.01)结论冠心舒通胶囊可改善ISO诱导的大鼠心脏重塑,其机制可能与抗炎、抑制TGF-β/Smad信号通路有关。

Objective To investigate the inhibitory effect of Guanxin Shutong capsule on cardie remodeling induced by isoproterenol (iso) via TGF-β/Smad signaling pathway.Methods SD rats were randomly divided into control group,model group,Guanxin Shutong capsule low,middle and high (1.2,2.4,4.8 g·kg^-1,respectively) dose groups,propranolol (0.5 g·kg^-1) group.The cardie remodeling model was established by subcutaneous injection of ISO (5 mg·kg^-1) for 7 days.From the second day of the start of modeling,drugs were given continuously for a month.The structure of myocardium was observed by echocardiography.EF,FS,EDV,ESV,LVIDd,LVPWd were measured;HMI,LVMI were measured and calculated.The morphological changes of myocardium were observed by Masson staining;biochemical method were used to detect the content of Hydro;the levels of serum interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α) were measured by ELISA;protein expression levels of TGF-β,Smad2/3,Caspase-3 were detected by Western Blot.Results Compared with the control group,the model group rats had decreased EF,FS,and increased EDV,ESV,LVIDd,LVPWd,HMI,LVMI.Cardiac Masson staining showed severe fibrosis;CVF,Hydro,serum contents of TNF-α and IL-6 were significantly increased,as well as the TGF-β,Smad2/3,Caspase-3 levels up-regulated (P < 0.01).Compared with the model group,the cardiac function was improved in the Guanxin Shutong capsule middle and high dose groups;the myocardial fibrosis level and the content of Hydro were decreased;levels of TNF-α and IL-6 were decreased;expressions of TGF-β,Smad2/3,Caspase-3 were decreased at the same time (P < 0.05,P < 0.01).Conclusion Guanxin Shutong capsule has a protective effect on isoproterenol-induced cardie remodeling,which is partially via inhibiting the TGF-β/Smad signaling pathway.