山药苦瓜复方制剂降低2型糖尿病大鼠血糖和血脂的实

背景药食两用中药山药、苦瓜和甘蔗纤维等降糖和降脂作用明显,但作用机制仍不十分清楚.目的观察山药苦瓜复方制剂对2型糖尿病大鼠血糖、血脂、血胰岛素水平及抗感染能力的影响.设计随机对照动物实验.单位广东医学院科技开发中心、药理教研室、实验动物中心、中心实验室.材料选用SPF级雌性SD大鼠80只,鼠龄4个月.二甲双胍(吉林省东北亚药业股份有限公司,批号040126).总胆固醇、高密度脂蛋白胆固醇、三酰甘油试剂盒购于北京中生生物技术股份有限公司;Surestep Life scan血糖仪及试纸(美国强生公司);胰岛素放射免疫试剂盒(上海海军医学研究所);UV-3010紫外分光光度计(日本岛津);山药苦瓜复方制剂由广东医学院科技开发中心提供,批号040321,混悬液,由山药、苦瓜和甘蔗渣膳食纤维等成分经水提而成,其中生药含量1 kg/L.方法实验中的动物饲养和标本采集于2004-06/2005-12在广东医学院实验动物中心和科技开发中心实验室完成,指标的检测在广东医学院药理研究室和中心实验室完成.①随机选取20只为正常对照组,每天灌胃生理盐水5 mL/kg.其余60只大鼠灌胃脂肪乳剂5 mL/kg,1次/d,连续4周后,大鼠禁食12 h,腹腔注射2g/L链脲佐菌素30 mg/kg.正常对照组腹腔注射等体积柠檬酸缓冲液.3 d后测随机血糖,2周后重复测血糖,以血糖水平高于13.5 mmol/L或尿糖＞++以上且持续2周为造模成功(n=48).②将48只大鼠按随机摸球法分为3组模型组,二甲双胍组和山药苦瓜复方制剂组,每组16只.模型组灌胃脂肪乳剂5 mL/kg;二甲双胍组和山药苦瓜复方制剂组灌胃脂肪乳剂5 mL/kg后分别灌胃二甲双胍1 mg/kg和含生药1 kg/L的山药苦瓜复方制剂5 mL/kg.正常对照组灌胃生理盐水5 mL/kg.各组均1次/d,连续6周.③在实验结束前1天测随机血糖.禁食12 h用放射免疫法测定血浆胰岛素水平,用分光光度法测定血浆总蛋白、白蛋白水平,按相应试剂盒说明测定三酰甘油、总胆固醇、高密度脂蛋白胆固醇水平.④计量资料差异比较采用方差分析,先用Levene'stest对数据进行方差齐性检验,如方差齐用Bonferroni检验,如方差不齐则用Tamhane's T2检验.主要观察指标山药苦瓜复方制剂对2型糖尿病大鼠血糖、胰岛素、血脂和血浆蛋白水平的影响.结果造模失败脱失12只,模型组和二甲双胍组在实验过程有4,2只动物死亡,最终62只进入结果分析.①空腹血糖和血浆胰岛素测定结果正常对照组大鼠空腹血糖水平明显低于其他3组(t=2.673～4.224,＜0.05～0.01),血胰岛素水平明显高于其他3组(t=3.780～5.824,P＜0.05～0.01).模型组大鼠空腹胰岛素水平明显低于山药苦瓜复方制剂组(t=2.825,P＜0.05),空腹血糖水平明显高于二甲双胍组和山药苦瓜复方制剂组(t=3.906,3.056,P＜0.05).二甲双胍组大鼠胰岛素水平明显低于山药苦瓜复方制剂组(t=3.014,P＜0.05),但空腹血糖水平与山药苦瓜复方制剂组相近(P＞0.05).②血脂水平测定结果正常对照组大鼠血浆总胆固醇和三酰甘油水平明显低于其他3组,高密度脂蛋白胆固醇水平明显高于其他3组(t=2.521～4.892,P＜0.05～0.01).模型组大鼠血浆总胆固醇水平明显高于二甲双胍组和山药苦瓜复方制剂组(t=2.466,2.512,P＜0.05),三酰甘油水平明显高于山药苦瓜复方制剂组(t=2.612,P＜0.05),高密度脂蛋白胆固醇水平明显低于山药苦瓜复方制剂组(t=3.688,P＜0.05).二甲双胍组大鼠血浆三酰甘油水平明显高于山药苦瓜复方制剂组(t=2.620,P＜0.05).③血浆蛋白水平测定结果各组大鼠血浆总蛋白水平相近(P＞0.05);正常对照组大鼠血浆白蛋白水平明显高于模型组和二甲双胍组(t=3.773,3.104,P＜0.05),与山药苦瓜复方制剂组相近(P＞0.05),白蛋白与球蛋白比值明显高于其他3组(t=2.830～3.056,P＜0.05).模型组大鼠血浆白蛋白水平和白蛋白与球蛋白比值明显低于山药苦瓜复方制剂组(t=2.604,3.808,P＜0.05).结论山药苦瓜复方制剂可降低2型糖尿病模型大鼠血糖、血脂水平,升高胰岛素水平,提高抗感染能力.

Effect of compound preparation of common yam rhizome and balsampear fruit on blood glucose and lipid in rats with type 2 diabetes mellitus

BACKGROUND: Chinese herbs of common yam rhizome, balsampear fruit and bagasse fiber have good effects on decreasing blood glucose and lipid, but its mechanism is unclear.OBJECTIVE: To observe the effect of compound preparation of common yam rhizome and balsampear fruit on blood glucose, lipid, blood insulin and anti-infection of rats with type 2 diabetes mellitus (DM).DESIGN: Randomized controlled animal study.SETTING: Technological Developing Center, Pharmacological Department, Experimental Animal Center, and Central Laboratory of Guangdong Medical College.MATERIALS: A total of 80 female SD rats with 4 months old and of SPF grade were selected in this study. Flumamine (Jilin Dongbeiya Pharmaceutical Co., Ltd., batch number: 040126); total cholesterol (TC), high density lipoprotein cholesterol (HDL-C) and triacylglycerol (TG) (Beijing Zhongsheng Biotechnological Co. Ltd.); Surestep Life scan blood glucose meter and test paper (Johnson Company, USA); insulin radio-immunity kit (Shanghai Navy Medical Institute); UV-3010 ultraviolet spectrophotometer (Japan Shimadzu); compound preparation of common yam rhizome and balsampear fruit (Technological Developing Center of Guangdong Medical College, batch number: 040321); suspension was extracted from common yam rhizome, balsampear fruit and dietary fiber of bagasse through water with 1 kg/L raw materials.METHODS: Animal breeding and samples collecting were carried out in the Experimental Animal Center and Technological Developing Central Laboratory of Guangdong Medical College from June 2004 to December 2005; meanwhile, detection of marker was carried out in the Pharmacological Department and Central Laboratory of Guangdong Medical College. ①Twenty rats were randomly selected as normal control group and perfused with 5 mL/kg saline every day. Other 60 rats were perfused with 5 mL/kg fat emulsion once a day for 4 weeks, and then, rats were fasted for 12hours and peritoneally injected with 2 g/L streptozotocin (30 mg/kg). Rats in normal control group were peritoneally injected with the same volume of citromalic acid buffer. Three days later, blood glucose was measured ranomly and measured again after 2 weeks. If level of blood glucose was igher than 13.5 mmol/L or urinal glucose was ＞ ++ for two weeks, the models were successful (n=48). ② According to random lot method, 48 rats were divided into three groups: model group, flumamine group and comound preparation group with 16 in each group. Rats in model group were perfused with 5 mL/kg fat emulsion; moreover, rats in flumamine groupr and compound preparation group were perfused with 5 mL/kg fat emulsion and then with 1 mg/kg flumamine and 5 mL/kg compound preparation including 1 kg/L raw drug, respectively. Rats in normal control group were perfused with 5 mL/kg saline. All rats in each group were perfused once a day for 6 weeks in total. ③ Value of blood glucose was measured at one day before the experiment finished. Twelve hours after fasting, level of plasma insulin was measured with radio-immunity method; levels of plasma total protein and albumin were measured with spectrophotography; levels of TG, TC and HDL-C were measured with the related kits. ④ Measurement data were compared with analysis of variance (ANOVA). Levene's test was firstly used to evaluate regularity of variance. Bonferroni test was used for regular variance; however, Tamhane's T2 was used.MAIN OUTCOME MEASURES: Effect of compound preparation on levels of blood glucose, insulin, lipid and plasma protein of rats with type 2 DM.RESULTS: Twelve rats were lost because of failure in modeling, and 4rats in model group and 2 in flumamine group died during the experiment,respectively. Therefore, 62 rats were involved in the final analysis. ①Measurement of fasting blood glucose and plasma insulin: Value of fasting blood glucose in normal control group was lower than that in other three groups (t=2.673-4.224, P ＜ 0.05-0.01), but level of plasma insulin was higher than that in other three groups (t=3.780-5.824, P ＜ 0.05-0.01).Fasting insulin in model group was lower than that in compound prepara tion group (t=2.825, P ＜ 0.05); fasting blood glucose was higher than that in flumamine group and compound preparation group (t=3.906, 3.056, P ＜ 0.05); * level of insulin in flumamine group was lower than that in compound preparation group (t=3.014, P ＜ 0.05); level of fasting blood glucose in flumamine group was close to that in compound preparation group (P ＞ 0.05). ② Measurement of lipid: Levels of TC and TG in normal control group were lower than those in other three groups, but level of HDL-C was higher than that in other three groups (t=2.521-4.892, P ＜ 0.05-0.01).Plasma TC in model group was higher than that in flumamine group and compound preparation group (t=2.466-2.512, P ＜ 0.05), value of TG was higher than that in compound preparation group (t=2.612, P ＜ 0.05), and level of HDL-C was lower than that in compound preparation group (t =3.688, P ＜ 0.05). Plasma TG in flumamine group was higher than that in compound preparation group (t=2.620, P ＜ 0.05). ③ Measurement of plasma protein: Levels of plasma total protein were close to each other (P ＞ 0.05). Plasma albumin in normal control group was higher than that in model group and flumamine group (t=3.773, 3.104, P ＜ 0.05), but that was close to that in compound preparation group (P ＞ 0.05). Ratio between albumin and globulin in normal oln that in other groups (t=2.830-3.056, P ＜ 0.05). Level of plasma albumin and ratio between albumin and globulin were lower in model group than those in compound preparation group (t=2.604, 3.808, P ＜ 0.05).CONCLUSION: Compound preparation can decrease levels of blood glucose and lipid, increase content of insulin, and improve anti-infection ability of rats with type 2 DM.