汉黄芩素调节Hippo/YAP信号通路对慢性心力衰竭大鼠心肌纤维化的影响

Effect of Wogonin on Myocardial Fibrosis in Rats with Chronic Heart Failure by Regulating Hippo/YAP Signaling Pathway

目的:探究汉黄芩素调节Hippo/Yes相关蛋白(YAP)信号通路对慢性心力衰竭(CHF)大鼠心肌纤维化的影响。方法:采用腹腔注射阿霉素的方式构建大鼠CHF模型,成功造模36只大鼠,将其随机分为模型组、汉黄芩素组(汉黄芩素100 mg/kg)、汉黄芩素+维替泊芬组(100 mg/kg汉黄芩素+100 mg/kg维替泊芬),每组12只。剩余12只大鼠作为对照组。采用彩色多普勒超声系统检测各组大鼠心功能参数:左室射血分数(LVEF)、左室短轴缩短率(LVFS)、左室舒张末期内径(LVEDD)、左室收缩末期内径(LVESD);称重并计算各组大鼠心脏质量指数(HMI)和左心室质量指数(LVMI);酶联免疫吸附法(ELISA)检测心肌组织超氧化物歧化酶(SOD)、丙二醛(MDA)水平;Masson染色法检测大鼠心肌胶原容积分数(CVF)和血管周围纤维面积和血管管腔面积比值(PVCA/LA);苏木精-伊红(HE)染色观察大鼠心肌组织病理学变化;蛋白免疫印迹(Western Blot)法检测大鼠心肌组织Hippo/YAP信号通路相关蛋白、基质金属蛋白酶(MMP)-2、MMP-9、结缔组织生长因子(CTGF)表达。结果:与对照组相比,模型组大鼠LVEF、LVFS、心肌组织SOD水平及哺乳动物ste-20样激酶(MST)1/2、大型肿瘤抑制因子(LATS)1/2磷酸化水平明显降低(P<0.05),LVEDD、LVESD、HMI、LVMI、心肌组织MDA水平、CVF、PVCA/LA、心肌组织YAP、MMP-2、MMP-9、CTGF蛋白表达明显升高(P<0.05),心肌组织出现大量胶原纤维沉积和损伤。与模型组相比,汉黄芩素组LVEF、LVFS、心肌组织SOD水平、MST1/2、LATS1/2磷酸化水平明显升高(P<0.05),LVEDD、LVESD、HMI、LVMI、心肌组织MDA水平、CVF、PVCA/LA、心肌组织YAP、MMP-2、MMP-9、CTGF蛋白表达明显降低(P<0.05),心肌组织胶原纤维沉积和损伤减少。维替泊芬增强了汉黄芩素对CHF大鼠心肌纤维化的改善作用。结论:汉黄芩素可能通过激活Hippo/YAP信号通路、抑制YAP表达来减轻CHF大鼠心肌纤维化。

Objective:To explore the effect of Wogonin on myocardial fibrosis in rats with chronic heart failure(CHF)regulating Hippo/Yes associated protein(YAP)signaling pathway.Methods:The rat CHF model was established by intraperitoneal injection of adriamycin.Thirty-six rats were successfully modeling and randomly divided into model group,wogonin group(100 mg/kg baicalein)and wogonin+vetipofen group(100 mg/kg baicalein+100 mg/kg vetipofen),with 12 rats in each group.The remained 12 rats served as the control group.Left ventricular ejection fraction(LVEF),left ventricular brachyaxis shortening rate(LVFS),left ventricular end-diastolic diameter(LVEDD)and left ventricular end-systolic diameter(LVESD)were measured by color Doppler ultrasound.Heart mass index(HMI)and left ventricular Mass index(LVMI)were detected.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in myocardial tissue were detected by enzyme-linked immunosorbent assay(ELISA).Masson staining was used to detect myocardial collagen volume fraction(CVF)and the ratio of perivascular fiber area to vascular lumen area(PVCA/LA).Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of rat myocardium.The expressions of Hippo/YAP signaling pathway related proteins,matrix metalloproteinase(MMP-2),MMP-9 and connective tissue growth factor(CTGF)were detected by Western Blot.Results:Compared with the control group,LVEF,LVFS,SOD in myocardial tissue and the phosphorylation levels of STE-20-like kinase(MST)1/2 and large tumor suppressor factor(LATS)1/2 in the model group significantly decreased(P<0.05),LVEDD,LVESD,HMI,LVMI,MDA,CVF,PVCA/LA,YAP,MMP-2,MMP-9,and CTGF protein expressions in myocardial tissue significantly increased(P<0.05),and a large number of collagen fiber deposition and injury appeared in myocardial tissue.Compared with the model group,LVEF,LVFS,SOD,MST1/2 and LATS1/2 phosphorylation in myocardial tissue in the wogonin group significantly increased(P<0.05),LVEDD,LVESD,HMI,LVMI,MDA,CVF,PVCA/LA,YAP,MMP-2,MMP-9,and CTGF protein expressions in myocardial tissue significantly decreased(P<0.05),and collagen fiber deposition and injury in myocardial tissue decreased.Vitipofen enhanced the ameliorative effect of wogonin on myocardial fibrosis in CHF rats.Conclusion:Wogonin might reduce myocardial fibrosis in CHF rats by activating Hippo/YAP signaling pathway and inhibiting the expression of YAP.