鬼针草总黄酮调控PI3K/Akt/mTOR自噬通路对高血压大鼠心肌纤维化的影响  

Effects of total flavonoids of Stipium bidens bipinnata L on myocardial fibrosis in hypertensive rats by regulating autophagy pathway of PI3K/Akt/mTOR

作　　者：董凤梅 韩凌 DONG Feng-mei;HAN Ling(Department of Internal Medicine-Cardiovascular,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,Henan,China)

机构地区：[1]郑州大学附属郑州中心医院心内科,河南郑州450007

出　　处：《广东医学》2023年第6期720-728,共9页Guangdong Medical Journal

基　　金：河南省医学科技攻关省部共建重点项目(SBGJ202102152)。

摘　　要：目的探讨鬼针草总黄酮(TFB)调控磷脂肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/雷帕霉素靶蛋白(mTOR)自噬通路对自发性高血压(SHR)大鼠心肌纤维化(MF)的影响。方法将30只SHR大鼠随机分为5组:SHR组、SHR+坎地沙坦组、SHR+TFB组、SHR+IGF-1(PI3K/Akt/mTOR通路激活剂)组、SHR+TFB+IGF-1组,每组6只。另取6只年龄匹配的雄性WKY大鼠作为WKY大鼠对照组。WKY大鼠对照组和SHR组大鼠予剂量匹配的生理盐水灌胃,SHR+坎地沙坦组予溶于生理盐水的坎地沙坦(40 mg/kg)灌胃,SHR+TFB组予溶于生理盐水的TFB(240 mg/kg)灌胃,SHR+IGF-1组大鼠经侧脑室注射20μg/kg IGF-1,SHR+TFB+IGF-1组大鼠经侧脑室注射20μg/kg IGF-1后予240 mg/kg TFB灌胃,每日1次,连续12周。给药结束后,检测各组大鼠血压、左心室功能指标;HE和Masson染色观察组织病理变化;TUNEL染色观察组织细胞凋亡;试剂盒法测量抗氧化状态;RT-qPCR检测Ⅰ型胶原蛋白(CollagenⅠ)、α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(Fibronectin)和波形蛋白(Vimentin)mRNA水平;Western blot检测CollagenⅠ、α-SMA、Fibronectin、Vimentin、自噬标志蛋白(Beclin-1、LC3B、p62)、PI3K/Akt/mTOR信号通路蛋白水平。结果与SHR组相比,SHR+坎地沙坦组、SHR+TFB组大鼠左室射血分数(EF)、左室短轴缩短率(FS)明显升高,收缩压、左室舒张末期内径(LVEDD)、左室舒张末期容积(LVEDV)、左室收缩末期内径(LVESD)、左室收缩末期容积(LVESV)、左心室质量指数(LVMI)明显降低,MF程度减轻,CollagenⅠ、α-SMA、Fibronectin、Vimentin表达显著下调,凋亡小体百分比显著降低,谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)显著升高,内源性过氧化氢(H_(2)O_(2))、丙二醛(MDA)和活性氧(ROS)显著降低,Beclin-1、LC3B-Ⅱ/LC3B-Ⅰ显著升高,p62表达显著降低,p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR比值显著降低(P<0.05)。与SHR+TFB组相比,SHR+TFB+IGF-1组EF、FS明显降低,收缩压、LVEDD、LVEDV、LVESD、LVObjective To investigate the effects of total flavonoids of Stipium bidens bipinnata L(TFB)on myocardial fibrosis(MF)in spontaneously hypertensive(SHR)rats by regulating the autophagy pathway of phosphoinositide-3 kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR).Methods Thirty SHR rats were randomly divided into 5 groups,SHR group,SHR+candesartan group,SHR+TFB group,SHR+IGF-1(PI3K/Akt/mTOR pathway activator)group and SHR+TFB+IGF-1 group(n=6),and 6 age-matched male WKY rats were enrolled as control group.After the medication was administered,systolic blood pressure and left ventricular function indicators of rats were measured.Histopathological changes were observed by HE and Masson staining.Tissue cell apoptosis were observed by TUNEL staining.Antioxidant status were measured by kit method.The mRNA levels of CollagenⅠ,α-SMA,Fibronectin,Vimentin were measured by RT-QPCR.The levels of CollagenⅠ,α-SMA,Fibronectin,Vimentin,autophagy marker proteins(Beclin-1,LC3B,p62)and PI3K/Akt/mTOR signaling pathway proteins were measured by Western Blot.Results Compared with SHR group,EF and FS in SHR+TFB group were significantly increased;systolic blood pressure,LVEDD,LVEDV,LVESD,LVESV and LVMI were significantly reduced;MF degree was significantly alleviated;CollagenⅠ,α-SMA,Fibronectin and Vimentin expressions were significantly down-regulated;the percentage of apoptotic bodies was significantly reduced;GSH-Px and SOD were significantly increased;H_(2)O_(2),MDA and ROS were significantly reduced;Beclin-1 and LC3B-Ⅱ/LC3B-Ⅰwere significantly increased;p62 was significantly reduced;and the ratios of p-PI3K/PI3K,p-Akt/Akt and p-mTOR/mTOR were significantly reduced(P<0.05).Compared with SHR+TFB group,EF and FS in SHR+TFB+IGF-1 group were significantly decreased;systolic blood pressure,LVEDD,LVEDV,LVESD,LVESV and LVMI were significantly increased;MF degree was significantly aggravated;CollagenⅠ,α-SMA,Fibronectin and Vimentin expressions were significantly up-regulated;the percentage of apoptotic b

关 键 词：鬼针草总黄酮 PI3K/Akt/mTOR自噬通路 高血压 心肌纤维化 

分 类 号：R544.1[医药卫生—心血管疾病] R285.5[医药卫生—内科学]