松果菊苷对db/db糖尿病小鼠心肌细胞脂质代谢紊乱的改善作用  

Protective Effect of Echinacoside on Lipid Metabolism Disorders in Cardiomyocytes of db/db Diabetic Mice

作　　者：张祥 杨洛 廖敏 郝亚荣[1] Zhang Xiang;Yang Luo;Liao Min(Renmin Hospital of Wuhan University,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》2020年第4期87-92,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：目的探究肉苁蓉提取物松果菊苷(echinacoside)对糖尿病心肌病db/db小鼠模型心肌细胞糖脂代谢紊乱是否具有保护作用及可能作用机制。方法将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,按照数字表法随机取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)和松果菊苷干预组(db/db+ECH组,n=10)。其中db/db组和db/m组每天按照0.05ml/10g标准进行0.9%NaCl溶液灌胃10周。db/db+ECH组小鼠按松果菊苷300mg/(kg·d)灌胃干预10周。每周监测各组小鼠体质量,观察小鼠摄食、饮水及活动情况。于小鼠第18周龄用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(FBG);分离血清检测各组小鼠血清脂质水平变化;采用HE染色观察心肌组织病理改变;苏丹红Ⅱ染色观察心肌细胞脂质蓄积情况;免疫印迹(Western blot)法检测糖脂代谢相关蛋白PPAR-α、CPT-Ⅰ、CD36和GLUT-4的表达水平;real-time PCR测定心肌细胞PPAR-α、CPT-Ⅰ、CD36和GLUT-4 mRNA的表达水平。结果与db/m组比较,db/db组小鼠体质量、血糖和血脂水平显著升高(P<0.05);同时HE染色切片显示,与db/m组比较,db/db组小鼠心肌细胞可见明显的肥大、坏死,心肌细胞排列结构紊乱;而db/db+ECH组小鼠心肌细胞形态有所缓解,细胞间隙减少,排列较规则。苏丹红Ⅱ染色观测心肌细胞脂质蓄积结果提示相较于db/m组,db/db组小鼠心肌细胞发生严重的脂质变性,可见不同大小的脂质液泡,同时伴有PPAR-α、CPT-Ⅰ和GLUT-4蛋白表达量显著降低(P<0.01),CD36蛋白表达量显著增加(P<0.01),且PPAR-α、CPT-Ⅰ、GLUT-4 mRNA表达量明显降低,CD36 mRNA水平上调(P<0.01)。与db/db组比较,db/db+ECH组上述指标均有明显逆转,且切片观察小鼠心肌细胞脂质沉积显著减少。结论松果菊苷能够明显改善db/db小鼠心肌细胞糖脂代谢紊乱,其机制可能与通过调控PPAR-α/CPT-ⅠObjective To investigate the protective effect and possible mechanism of echinacoside(ECH)on lipid metabolism disorders in cardiomyocytes of db/db diabetic mice.Methods Twenty healthy male db/db mice aged 6 weeks and weighing from 40g to 48g were divided into db/db+ECH group[n=10,ECH,300mg/(kg·d)]and db/db group(n=10,given the corresponding vplume of saline)by random number table.db/m mice were served as normal control group,that was db/m group(n=9,given the corresponding volume of saline).Three groups of mice were monitored weekly to observe the diet,water and activity of the mice.The mice were injected with 2%of pentobarbital sodium in week 10 and executed.The weight of mice was monitored weekly and the free blood glucose(FBG)was measured.HE staining and SudanⅡstaining were used to evaluate the myocardial tissue pathological changes and lipid accumulation in cardiomyocytes.Western blot method was uesd to detect the related protein expression of PPAR-α,CPT-Ⅰ,CD36 and GLUT-4.Total RNA of the myocardial cell was extracted and the mRNA of PPAR-α,CPT-Ⅰ,CD36 and GLUT-4 were detected by real-time quantitative PCR.Results Compared with the db/m group,the weight,blood glucose and serum lipid levels in the db/db group were significantly increased(P<0.05).Moreover,HE staining showed that compared with the db/m group,cardiomyocytes in the db/db group showed obvious hypertrophy,necrosis and disordered arrangement of cardiomyocytes.However,the morphology of cardiomyocytes in the db/db+ECH group was somewhat relieved,with reduced intercellular space and regular arrangement.SudanⅡmyocardial cell lipid accumulation observations showed that compared with the db/m group,db/db mice cardiomyocytes serious lipid degeneration,the lipid vacuoles of different sizes,accompanied by PPAR-α,CPT-Ⅰand GLUT-4 protein expression significantly reduced(P<0.01),the amount of CD36 protein expression significantly increased(P<0.01),and PPAR-α,CPT-Ⅰ,GLUT-4 mRNA expression quantity to decrease,CD36 mRNA level raised(P<0.01).Compared

关 键 词：糖尿病心肌病 松果菊苷 糖脂代谢紊乱 PPAR-Α DB/DB小鼠 

分 类 号：R587[医药卫生—内分泌]