松果菊苷对db/db糖尿病心肌病小鼠心肌细胞凋亡的改善作用  

Effects of Echinacoside on Apoptosis of Myocardial Cells in db/db Diabetic Cardiomyopathy Mice

作　　者：张祥 郝亚荣[1] ZHANG Xiang;HA0 Yarong(Department of Geriatrics,Renmin Hospital of Wuhan Uninersity,Hubei 430060,China)

机构地区：[1]武汉大学人民医院老年病科,430060

出　　处：《医学研究杂志》2023年第8期88-93,共6页Journal of Medical Research

基　　金：湖北省自然科学基金资助项目(面上项目)(2016CFB673)。

摘　　要：目的探讨肉苁蓉提取物松果菊苷(echinacoside,ECH)对db/db糖尿病心肌病小鼠心肌细胞的保护作用及其作用机制。方法选取29只SPF级6周龄雄性db/db小鼠和db/m小鼠适应性饲养1周后,按照随机数字表法取9只db/m小鼠作为正常对照组(db/m组,n=9),将db/db小鼠分为糖尿病心肌病组(db/db组,n=10)与ECH干预组(db/db+ECH组,n=10)。其中db/db组和db/m组小鼠每日按照0.05ml/10g标准行0.9%氯化钠溶液灌胃10周。db/db+ECH组小鼠按ECH 300mg/(kg·d)灌胃10周。每周监测3组的小鼠体质量,观察小鼠摄食、饮水及活动情况。于鼠龄第18周用2%戊巴比妥钠腹腔注射麻醉小鼠并处死,取尾静脉血测定空腹血糖和血脂。采用苏木精-伊红(hematoxylin eosin,HE)染色观察心肌组织病理改变,采用TUNEL染色法测定心肌细胞凋亡率,采用Western blot法检测凋亡相关蛋白p53、p38MAPK、caspase-3和caspase-8的表达水平,采用实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)测定心肌细胞p53m RNA、p38MAPK mRNA和caspase mRNA、Bcl-2mRNA、Bax mRNA的表达水平。结果与db/m组比较,db/db组小鼠的体质量、血糖和血脂水平显著升高(P<0.05或P<0.01)。HE染色结果显示,与db/m组比较,db/db组小鼠的心肌细胞可见明显肥大、坏死,心肌细胞排列结构紊乱;而db/db+ECH组小鼠的心肌细胞形态有所缓解,细胞间隙减少,排列较规则。TUNEL染色结果显示,与db/m组比较,db/db组小鼠的心肌细胞凋亡程度显著增高(P<0.01),同时伴有p53蛋白、p38MAPK蛋白、caspase-3和caspase-8蛋白表达量显著升高(P<0.01),且p53mRNA、p38MAPK mRNA和caspase mRNA、Bax mRNA表达量明显上升,Bcl-2mRNA水平下调(P<0.01)。与db/db组比较,db/db+ECH组小鼠上述指标均有明显降低,且TUNEL染色观察小鼠心肌细胞凋亡指数显著下降(P<0.01)。结论ECH能够明显改善db/db小鼠心肌细胞凋亡,其机制可能与通过p53/p38MAPK/caspase信号通路有关。Objective To investigate the protective effect of echinacoside(ECH)extracted from cistanche deserticola on myocardial cells of db/db diabetic cardiomyopathy mice and its possible mechanism.Methods Twenty-nine SPF 6-week-old male db/db mice and db/m mice were adapted to feeding for 1 week.According to the random number table method,9db/m mice were selected as the normal control group(db/m group,n=9),and db/db mice were divided into the diabetic cardiomyopathy group(db/db group,n=10)and the ECH intervention group(db/db+ECH group,n=10).The mice in the db/db group and the db/m group were given normal saline daily for 10 weeks according to the standard of 0.05ml/10g.The mice in the db/db+ECH group were given ECH 300mg/(kg·d)by gavage for 10 weeks.The weight of mice in three groups was monitored every week,and the food intake,water intake and activity of mice were observed.The mice were anesthetized by intraperitoneal injection of 2%pentobarbital sodium at 18th week of age and sacrificed.Tail venous blood was taken to measure fasting blood glucose and blood lipid.Hematoxylin eosin(HE)staining was used to observe the pathological changes of myocardial tissue,TUNEL staining was used to detect the apoptosis rate of myocardial cells,Western blot was used to detect the expression levels of apoptosis-related proteins p53,p38MAPK,caspase-3 and caspase-8,and real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression levels of p53mRNA,p38MAPK mRNA,caspase mRNA,Bcl-2mRNA and Bax mRNA.Resultss Compared with the db/m group,the body weight,blood glucose and blood lipid levels of the db/db mice were significantly increased(P<0.05 or P<0.01).HE staining showed that,compared with the db/m group,the myocardial cells in the db/db group showed obvious hypertrophy,necrosis and disordered arrangement of myocardial cells.However,the morphology of myocardial cells in db/db+ECH group was significantly improved,the intercellular space was reduced,and the arrangement was more regular.TUNEL staining showed that c

关 键 词：松果菊苷 糖尿病心肌病 凋亡 P53 P38MAPK 

分 类 号：R542.2[医药卫生—心血管疾病]