丹瓜方对ApoE^-/-糖尿病模型小鼠糖脂代谢及血管细胞黏附分子-1及mRNA表达水平的影响  

Effect of Dangua Recipe on Glycolipid Metabolism and VCAM-1 and Its mRNA Expression Level in Apo E^(-/-)Mice with Diabetes Mellitus

作　　者：衡先培[1,2] 李亮[3] 黄苏萍 陈燕[3] 林妙娴[3] 庄怀山[3] 颜群芳 杨柳清[1,2] 陈玲[5] 林青[1,2] 程心玲[1,2] 陈敏灵[1,2] 陈依楚 蓝元隆[3] 王志塔 姚淑红[3] 张治三 

机构地区：[1]福建中医药大学附属人民医院 [2]福建省人民医院内分泌科,福州350004 [3]福建中医药大学研究生部,福州350108 [4]福建中西医结合研究院,福州350108 [5]福建中医药大学生理学教研室,福州350108

出　　处：《中国中西医结合杂志》2014年第9期1086-1095,共10页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81173179);福建省自然科学基金资助项目(No.2011J01198);福建省科技社会发展重点项目(2012Y0037);陈可冀中西医结合发展基金资助项目(No.CKJ2009004)

摘　　要：目的研究丹瓜方对载脂蛋白E基因敲除(ApoE^(-/-))小鼠复合糖尿病模型糖脂代谢指标、血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)及其mRNA表达水平的影响,探讨丹瓜方防治糖尿病合并血管病变的部分机制。方法 Apo E^(-/-)小鼠皮下注射链脲佐菌素(STZ)制备糖尿病模型。将32只成模鼠按照体重分层,再按血糖水平以随机数字表法分为模型组[无菌水(SW),15 m L/(kg·d)]、丹瓜方组[丹瓜方(DG),15 m L/(kg·d)]、联合用药组[吡格列酮(PG),4.3 mg/(kg·d)加DG1 5 mL/(kg·d)]、吡格列酮组[PG,4.3 mg/(kg·d)],每组8只。选血糖正常的8只同龄同品系C 57小鼠作为对照组。灌胃干预12周。监测各组小鼠空腹血糖(FBG)、体重、摄食量及饮水量;测量皮温及鼠尾长度,并通过肉眼观察肝脂肪变程度;测定各组小鼠血清HbA1 c、TC、LDL-C水平,采用放射免疫法测定ET-1,生化法—硝酸还原酶法测定NO,E LISA法测定VCAM-1,应用实时荧光定量PCR(qPCR)检测肾组织VCAM-1 mRNA表达。结果与对照组比较,模型组小鼠体重及摄食量减少,饮水量增加(P<0.05),且血糖值曲线全程高于对照组(P<0.01)。与模型组比较,丹瓜方组小鼠体重增量、HbA1 c、TC、LDL-C、ET-1、VCAM-1水平显著降低,皮温升高(P<0.05或P<0.01)。与吡格列酮组比较,丹瓜方组小鼠体重、体重增量、FBG、TC及LDL-C水平降低(P<0.05或P<0.01)、饮水量和摄食量增加,鼠尾较长(P<0.01);各组小鼠NO水平比较,差异无统计学意义。与对照组比较,模型组肝脂肪变程度较重(P<0.05);与模型组及吡格列酮组比较,丹瓜方组脂肪变程度明显减轻(P<0.05,P<0.01),吡格列酮组肝脂肪变程度重于模型组(P<0.01),而联合用药组肝脂肪变程度处于丹瓜方组和吡格列酮组之间。丹瓜方组VCAM-1 mRNA的表达显著低于模型组(P<0.01),且低于联合用药组和吡格列酮组(P<0.05)。结论丹瓜方对自发性动脉粥样硬化和高脂血症的ApoE^(-/-)Objective To study the effect of Dangua Recipe（DGR） on glycolipid metabolism,vascular cell adhesion molecule-1（VCAM-1） and its mRNA expression level of transgenic Apo E^-/-mouse with spontaneous atherosclerosis,thus revealing its partial mechanism for curing diabetes mellitus（DM） with angiopathy.Methods Diabetic model was prepared by peritoneally injecting streptozotocin（STZ） to Apo E^-/-mouse.Totally 32 modeled mice were stratified by body weight,and then divided into 4 groups referring to blood glucose levels from low to high by random digit table,i.e.,the model group（MOD,fed with sterile water,at the daily dose of 15 mL/kg）,the DGR group（fed with DGR at the daily dose of 15 mL/kg）,the combination group（COM,fed with DGR at the daily dose of 15 mL/kg and pioglitazone at the daily dose of 4.3 mg/kg）,and the pioglitazone group（PIO,at the daily dose of 4.3 mg/kg）,8in each group.Another 8 normal glucose C57 mouse of the same age and strain were recruited as the control group.All interventions lasted for 12 weeks by gastrogavage.The fasting blood glucose（FBG）,body weight,food intake,water intake,skin temperature,the length of tail,and the degree of fatty liver were monitored.The hemoglobin A1c（HbA1c）,total cholesterol（TC）,and LDL-C were determined.Endothelin-1（ET-1） was determined by radioimmunoassay.Nitrogen monoxidum（NO） was determined by nitrate reductase.The kidney tissue VCAM-1 level was analyzed with ELISA.The expression of VCAM-1mRNA in the kidney tissue was detected with real time quantitative PCR.Results Compared with the control group,the body weight and food intake decreased,water intake increased in all the other model groups（P0.05）.Besides,the curve of blood glucose was higher in all the other model groups than in the control group（P〈0.01）.Compared with the model group,the body weight increased;levels of HbA1 c,TC,LDL-C,ET-1,and VCAM-1 were significantly lower;and skin temperature was higher in the DGR group（P〈0.05,P〈0.01）.Compared wit

关 键 词：丹瓜方 糖尿病 血管病变 高糖无害化 糖脂代谢 

分 类 号：R285.5[医药卫生—中药学]