侧柏叶乙酸乙酯提取物缓解小鼠糖尿病心肌病  

Ethyl acetate extract from Platycladus orientalis leaves alleviates diabetic cardiomyopathy in mice

作　　者：刘梦晴 肖子怡 黄依芳 刘文丽 谷雨 王叶凌 沈哲慧 李丽[1] LIU Mengqing;XIAO Ziyi;HUANG Yifang;LIU Wenli;GU Yu;WANG Yeling;SHEN Zhehui;LI Li(College of Pharmacy,Jining Medical University,Rizhao 276826,China)

机构地区：[1]济宁医学院药学院,山东日照276826

出　　处：《中国病理生理杂志》2024年第8期1417-1425,共9页Chinese Journal of Pathophysiology

基　　金：济宁医学院大学生创新创业项目(No.cx2022078z);山东省中医药科技项目(No.M-2023181)。

摘　　要：目的:研究侧柏叶乙酸乙酯提取物(EAEPOL)缓解糖尿病心肌病(DCM)的作用及其机制。方法:将健康成年C57BL/6小鼠随机分为正常对照组、DCM组和EAEPOL组。以小鼠心脏超声心动图为指标观察心脏结构和功能。以心肌羟脯氨酸含量、心肌组织Masson染色和天狼星红染色及心肌I型胶原(Col I)和III型胶原(Col III)免疫组化染色评价心肌纤维化程度。试剂盒检测心肌丙二醛(MDA)、超氧化物歧化酶(SOD)和总抗氧化能力(T-AOC),以及qRT-PCR和Western blot检测心肌血红素加氧酶1(HO-1)和核因子E2相关因子2(Nrf-2)表达水平,评价心肌氧化应激状态。Western blot检测CD31和α-平滑肌肌动蛋白(α-SMA)表达,qRT-PCR检测钙黏素5(CDH5)和成纤维细胞特异性蛋白1(FSP1)的mRNA水平,以及心肌石蜡切片α-SMA免疫荧光染色,评价心肌内皮-间充质转化(EndMT)程度。结果:(1)小鼠心脏超声心动图显示,与正常对照组相比,DCM组小鼠心率(HR)和射血分数(EF)显著下降(P<0.05或P<0.01),收缩期和舒张期左心室前壁厚度(LVAWs和LVAWd)及收缩期和舒张期左心室后壁厚度(LVPWs和LVPWd)均显著增大(P<0.05);与DCM组相比,EAEPOL组小鼠HR和EF显著增大(P<0.01);LVAWs、LVAWd、LVPWs和LVPWd显著减小(P<0.05)。(2)与正常对照组相比,DCM组小鼠心肌羟脯氨酸含量、天狼星红和Masson染色所示胶原面积比及Col Ⅰ和Col Ⅲ免疫组化阳性面积比均显著增加(P<0.01);与DCM组相比,EAEPOL组小鼠以上心肌纤维化指标均显著下降(P<0.05或P<0.01)。(3)与正常对照组相比,DCM组小鼠心肌MDA含量和Nrf-2表达显著增加,SOD活性、T-AOC水平和HO-1表达显著下降(P<0.01);与DCM组相比,EAEPOL组小鼠心肌MDA含量显著减少,SOD、T-AOC、Nrf-2和HO-1水平显著增加(P<0.05或P<0.01)。(4)与正常对照组相比,DCM组小鼠心肌CD31和CDH5的表达显著减弱,α-SMA和FSP1的表达显著增强(P<0.05或P<0.01),α-SMA免疫荧光阳性面积显著增大(P<0.01);与DCM组相比,EAIM:To investigate the alleviating effect of ethyl acetate extract from Platycladus orientalis leaves(EAEPOL)on diabetic cardiomyopathy(DCM)and its mechanisms.METHODS:Healthy adult C57BL/6 mice were randomly divided into normal control group,DCM group,and EAEPOL group.Cardiac structure and function of the mice were assessed by echocardiography.Myocardial fibrosis was evaluated though myocardial hydroxyproline content determination,myocardial Masson and Sirius red staining,and collagen type I(Col I)and collagen type III(Col III)immunohistochemistry.The degree of myocardial oxidative stress was assessed by measuring malondialdehyde(MDA),superoxide dismutase(SOD)and total antioxidative capacity(T-AOC)levels using kits,as well as detection of nuclear factor E2-related factor-2(Nrf-2)and heme oxygenase-1(HO-1)expression by qRT-PCR and Western blot.Endothelial-mesenchy mal transition(EndMT)was evaluated by detecting CD31 andα-smooth muscle actin(α-SMA)protein expression by Western blot,and cadherin 5(CDH5)and fibroblast specific protein 1(FSP1)mRNA expression by qRT-PCR,as well asα-SMA immunofluorescence staining.RESULTS:(1)Mouse echocardiography revealed that compared with normal control group,heart rate(HR)and ejection fraction(EF)were significantly reduced in DCM group(P<0.05 or P<0.01),while left ventricular anterior wall thickness at systole and diastole(LVAWs and LVAWd)and left ventricular posterior wall thickness at systole and diastole(LVPWs and LVPWd)were significantly increased(P<0.05).Compared with DCM group,the mice in EAEPOL group showed significant increases in HR and EF(P<0.01),and marked decreases in LVAWs,LVAWd,LVPWs and LVPWd(P<0.05).(2)Compared with normal control group,the content of hydroxyproline in mouse myocardium,the collagen area ratio shown by Sirius red and Masson staining,and the immunohistochemical positive area ratio of Col Ⅰ and Col Ⅲ in DCM group were significantly increased(P<0.01).Compared with DCM group,the above myocardial fibrosis indicators in EAEPOL group were significantl

关 键 词：侧柏叶乙酸乙酯提取物 糖尿病心肌病 心肌纤维化 氧化应激 内皮-间充质转化 

分 类 号：R587.1[医药卫生—内分泌] R541[医药卫生—内科学] R285.5[医药卫生—临床医学]