基于miRNA21-5p调控TGF-β1/Smads信号通路探讨苓桂气化方抗射血分数保留心力衰竭心肌纤维化的机制  

Exploring Anti-Myocardial Fibrosis Mechanism of Linggui Qihua Formula(苓桂气化方)Based on miRNA21-5p Regulating TGF-β1/Smads Pathway

作　　者：董国菊[1,2] 石玉姣 刘春秋 杨晨光 乔文博 刘永成 刘思雨 刘剑刚[2] DONG Guoju;SHI Yujiao;LIU Chunqiu;YANG Chenguang;QIAO Wenbo;LIU Yongcheng;LIU Siyu;LIU Jiangang(Xiyuan Hospital,Chinese Academy of Traditional Chinese Medicine,Beijing 100091,China;National Clinical Research Center for Chinese Medicine Cardiology,Xiyuan Hospital,Chinese Academy of Traditional Chinese Medicine,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院,北京100091 [2]中国中医科学院西苑医院国家中医心内科临床研究中心,北京100091

出　　处：《中华中医药学刊》2024年第4期1-6,I0001-I0005,共11页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074423);中国中医科学院科技创新重大攻关项目(C12021A00903)。

摘　　要：目的基于miRNA21-5p调控转化生长因子-β1(transforming growth factor beta 1,TGF-β1)/Smads信号通路探讨苓桂气化方抗射血分数保留心力衰竭(heart failure with preserved ejection fraction,HFpEF)心肌纤维化的机制。方法40只4周龄自发性高血压大鼠(spontaneously hypertensive rats,SHR)平均分为HFpEF组、沙库巴曲缬沙坦组(LCZ696,0.018 g·kg^(-1))、苓桂气化方低剂量组(LGQH-L,3.87 g·kg^(-1))和苓桂气化方高剂量组(LGQH-H,7.74 g·kg^(-1)),给予高脂、高盐及高糖饮食16周及腹腔注射链脲霉素溶液8周建立HFpEF大鼠模型。10只威斯塔京都(Wistar Kyoto,WKY)大鼠和10只SHR大鼠作为对照组,以普通饲料喂养至实验结束。造模成功后,WKY、SHR和HFpEF组给予等剂量生理盐水,其他3组按照预先规定的干预措施,每天灌胃1次,持续6周。干预结束后,行超声心动图测量左心室(left ventricle,LV)前壁厚度(LV end-diastolic anterior wall thickness,LVAWd)、LV后壁厚度(LV end-diastolic posterior wall thickness,LVPWd)、LV舒张末内径(LV end-diastolic internal diameter,LVIDd)、LV射血分数(LV ejection fraction,LVEF)、LV舒张早期二尖瓣流入峰值速度(E)、舒张晚期二尖瓣流入峰值速度(A)和LV舒张早期二尖瓣环运动速度(e'),并计算E/A和E/e';酶联免疫吸附试验检测血清心房钠尿肽(atrial natriuretic peptide,ANP)、B型利钠肽(B-type brain natriuretic peptide,BNP)及半乳糖凝集素3(Galectin-3,Gal-3);病理切片进行苏木精伊红及马松染色观察心肌肥厚及纤维化,并计算LV室壁厚度(LV wall thickness,LVWT)、胶原体积分数(collagen volume fraction,CVF)及血管周围纤维化比率(perivascular fibrosis ratio,PFR);实时定量聚合酶链反应检测LV心肌miRNA21-5p及TGF-β1/Smads信号通路相关mRNA表达;蛋白印迹检测LV心肌TGF-β1/Smads信号通路相关蛋白表达。结果与对照组比较,HFpEF组的LVAWd、LVPWd、LVIDd、E/A、E/e'、ANP、BNP、Gal-3、LVWT、CVF和PFR显著升高(P<0.05或P<0.0Objective To explore the anti-myocardial fibrosis mechanism of Linggui Qihua Formula(苓桂气化方,LGQH)based on miRNA21-5p regulating transforming growth factor-β1(TGF-β1)/Smads signaling pathway.Methods Forty 4-week-old spontaneously hypertensive rats(SHR)were equally divided into the heart failure with preserved ejection fraction(HFpEF)group,sacubitril/valsartan group(LCZ696,0.018 g·kg^(-1)),low dose LGQH group(LGQH-L,3.87 g·kg^(-1))and high dose LGQH group(LGQH-H,7.74 g·kg^(-1)).HFpEF rat model was established by feeding with the high-fat-salt-sugar diet for 16 weeks and intraperitoneal injection of streptozotocin solution for 8 weeks.10 Wistar Kyoto(WKY)rats and 10 SHRs were used as the control group and were given the regular feed until the end of the experiment.After successful molding,the WKY,SHR and HFpEF groups were given the equal dose of saline,and the other three groups followed the pre-specified intervention by gavage once a day for 6 weeks.After the intervention,echocardiography was performed to measure left ventricle(LV)end-diastolic anterior wall thickness(LVAWd),LV end-diastolic posterior wall thickness(LVPWd),LV end-diastolic internal diameter(LVIDd),LV ejection fraction(LVEF),diastolic mitral inflow peak velocity(E),late diastolic mitral inflow peak velocity(A),early diastolic mitral annular velocity(e')and E/A and E/e'were calculated.ELISA was used to detect serum atrial natriuretic peptide(ANP),B-type brain natriuretic peptide(BNP)and galectin-3(Gal-3).Hematoxylin eosin and Masson staining for pathology was used to visualize myocardial hypertrophy and fibrosis and calculate the LV wall thickness(LVWT),collagen volume fraction(CVF)and perivascular fibrosis ratio(PFR).The qPCR method was used to detect the expressions of miRNA21-5p and mRNA related to TGF-β1/Smads pathway in LV myocardium.Western blot was performed to detect the expression of protein related to TGF-β1/Smads pathway.Results Compared with the control group,the HFpEF group showed significantly higher levels of LVAWd,LVPW

关 键 词：miRNA21-5p TGF-β1/Smads信号通路 射血分数保留的心力衰竭 苓桂气化方 心肌纤维化 

分 类 号：R289.5[医药卫生—方剂学]