摘要
目的:观察具有化痰利湿、活血化瘀功效的二陈汤方加减干预2型糖尿病并发脂肪肝大鼠血糖、血脂、胰岛素的抵抗状况,以及脂肪肝模型病理形态学的变化。方法:实验于2004-04/11在厦门大学医学院动物实验中心完成。①选用健康Wistar大鼠40只,7周龄,雌雄各半。随机抽取12只为正常对照组,喂以普通饲料(其中各成分质量分数如下:碳水化合物0.6、蛋白质0.22,脂肪0.1,其他0.08,脂肪以豆油为主),其余则喂养高热量饲料(其中各成分质量分数如下:碳水化合物0.4,蛋白质0.13,脂肪0.4,其他0.07,脂肪以动物油脂为主)。2个月后,正常对照组的大鼠一次性腹腔注射0.1mol/LpH4.2枸椽酸缓冲液,其余28只大鼠则一次性腹腔注射链脲佐菌素25mg/kg。72h后查血糖≥16.7mmol/L为糖尿病大鼠,造模成功24只。②将24只糖尿病大鼠随机分为2组:模型组和二陈汤方加减组,每组12只。正常对照组喂以普通饲料,其余2组均喂养高热量饲料。二陈汤方加减组大鼠按8mL/(kg·d)剂量灌胃二陈汤方加减药物,由陈皮、半夏、茯苓、僵蚕及地龙等组成(上述中药由本院中药房提供),由本院药剂科制备成浓缩液(含生药2g/mL),1次/d;正常对照组和模型组灌胃等量的自来水,均连续干预16周。③然后采用ELISA法测血胰岛素水平,计算肝指数(肝湿重/体质量×100%)和胰岛素敏感指数ln｛[1/(空腹血糖×空腹胰岛素)]｝;光镜下评估肝脂肪变性和炎症坏死程度。炎症活动度计分[慢性肝炎炎症活动度及纤维化计分方案炎症活动度计分分为汇管区(P)、小叶内(L)、碎屑坏死(PN)及桥接坏死(BN)4项,每项依病变轻、中、重程度分别计以1,3,4分,计分公式为P+L+2(PN+BN)]。按照上海九强生物技术有限公司提供相应试剂盒说明书测定血清三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、谷草转氨酶、谷丙转氨酶、白蛋白及总蛋白水平。分别于造模前、造模后8周及造模后16周采用氧化酶法测定空腹血糖。④计量资料、等级资料、率差异比较分别采用两样本均数t检验、秩和检验、两样本率差别的统计意义检验。结果:由于造模失败脱失4只,最终进入结果分析36只。①肝指数:造模后16周模型组和二陈汤方加减组明显高于正常对照组(P<0.01)。②空腹血糖水平:造模后8和16周模型组及二陈汤方加减组均明显高于造模前和正常对照组(P<0.01)。造模后8和16周二陈汤方加减组明显低于模型组(P<0.05)。③血胰岛素水平:造模后16周模型组明显高于正常对照组和二陈汤方加减组(P<0.01)。④胰岛素敏感指数:造模后16周模型组明显低于正常对照组和陈汤方加减组(P<0.01)。⑤血清三酰甘油和低密度脂蛋白胆固醇水平:造模后16周,模型组和二陈汤方加减组明显高于正常对照组(P<0.01),二陈汤方加减组明显低于模型组(P<0.05)。⑥血清高密度脂蛋白胆固醇水平:造模后16周,模型组和二陈汤方加减组明显低于正常对照组(P<0.05),二陈汤方加减组明显高于模型组(P<0.05)。⑦血清谷丙转氨酶和谷草转氨酶活性:造模后16周,模型组和二陈汤方加减组明显高于正常对照组(P<0.05～0.01),二陈汤方加减组低于模型组,但差异不明显(P>0.05)。⑧肝脏炎症活动度计分:造模后16周,模型组及二陈汤方加减组明显高于正常对照组(P<0.01),但模型组与二陈汤方加减组差异不明显。⑨肝脏病理学形态变化:造模后16周,正常对照组大鼠肝脏无异常病变。模型组及二陈汤方加减组均出现弥漫性肝细胞脂肪变性,且两组肝脂变情况无区别。结论:二陈汤方加减可在一定程度上能改善糖尿病并发脂肪肝大鼠的血糖、血脂水平,胰岛素抵抗状况及肝功能指标,但不能减轻肝脏组织学病变。
AIM： To investigate effect of pingtang recipe characterized by resolving phlegm, removing damp by diuresis and promoting blood circulation by removing blood stasis, on blood glucose, lipid, resistance of insulin, liver function and pathomorphological changes of hepatic lipid in rats with type 2 diabetes mellitus （DM） complicated with fatty liver.
METHODS： The experiment was completed in the Animal Experimental Center in Medical College of Xiamen University from April to November of 2004. ① Totally 40 healthy Wistar rats aged 7 weeks old of half gender were selected in this study. Twelve rats were selected as normal control group and fed with ordinary diet （mass of main components： carbohydrate 0.6, protein 0.22, fat 0.1, others 0.08; soya bean oil especially in fat）, and other rats were fed with high-calorie diet （mass of main components： carbohydrate 0.4, protein 0.13, fat 0.4, others 0,07; animal lipid especially in fat）. After 2 months, rats in normal control group were injected with 0.1 mol/L pH4.2 citric-acid buffer solution and other 28 rats were injected with 25 mg/kg streptozotocin （STZ）. Blood glucose was tested after 72 hours, if fasting blood glucose （FBG） ≤ 16.7 mmol/L, the rats were diagrnosed as DM. So 24 rats successfully made into diabetic models.② Totally 24 diabetic rats were divided randomly into model group and pingtang recipe group with 12 in each group. Rats in normal group were fed with ordinary diet, but other rats were fed with high-calorie diet. Rats in pingtang recipe group were perfused with 8 mL/（kg·d） pingtang recipe once a day which consisted of chenpi, banxia, fuling, jiangchan and dilong provided by dispensary of our hospital and made into concintration of 2 g/mL raw materials. Rats in normal control group and model group were perfused with the same volume of water for 16 weeks. ③ Enzyme-linked immunoadsordent assay （ELISA） was used to assay levels of insulin, calculate hepatic index （hepatic wet weight/body mass × 100%） and sensitive index of insulin In [1/（fasting blood glucose × fasting insulin）]. Meanwhile, degeneration of hepatic fat and degrees of inflammatory necrosis were observed under light microscope. Scores of inflammatory activity： Inflammatory activity of chronic hepatitis and inflammatory activity of fibrosis were contained average area （P）, internal lobule （L）, debris necrosis （PN） and bridging necrosis （BN）. According to mild, moderate and severe degrees, each item was regarded as 1, 3 and 4 points as the following formula： P＋L＋2 （PN＋BN）. Contents of serum triacylglycerol （TG）, high density lipoprotein-cholesterol （HDL-C）, low density lipoproteincholesterol （LDL-C）, glutamic oxalaeetic transaminase （GOT）, glutamicpyruvic transaminase （GPT）, albumin and total protein were assayed with relevant kit provided by Shanghai Jiuqiang Biotechnology Company Limited. Fasting blood glucose was measured with oxidase method 8 weeks before modeling and 16 weeks after modeling. ④ Measurement data, ranked data and rate difference were compared with average t test, rank sum test and statistical test respectively.
RESULTS： Four rats failed to be models, 24 rats entered the final analysis.① Hepatic index in model and pingtang recipe group was higher than that in normal control group 16 weeks after modeling （P 〈 0.01）. ② Level of fasting blood glucose in model group and pingtang recipe group was higher 8 and 16 weeks after modeling than that before modeling and in normal control group （P 〈 0.01）, but that in pingtang recipe group was lower than that in model group 8 and 16 weeks after modeling （P 〈 0.05）. ③Level of blood insulin in model group was higher than that in normal control group and pingtang recipe group 16 weeks after modeling （P〈 0.01）. ④ Sensitive index of insulin in model group was lower than that in normal control group and pingtang recipe, group 16 weeks after modeling （P 〈 0.01）. ⑤ Levels of serum TG and LDL-C were higher in model group and pingtang recipe group than those in normal control group 16 weeks after modeling （P 〈 0.01）, but those in pingtang recipe group were lower than those in model group （P 〈 0.03）. ⑥ Level of HDL-C in model group and pingtang recipe group was lower than that in normal control group 16 weeks after modeling （P 〈 0.05）, but that in pingtang recipe group was higher than that in model group （P 〈 0.05）. ⑦ Activities of GPT and GOT in model group and pingtang recipe group were higher than those in normal control group 16 weeks after modeling （P 〈 0.05-0.01）, but those in pingtang recipe group were lower than those in model group, and there was not significantly different （P 〉 0.05）. ⑧ Scores of inflammatory activity in liver： Those in model group and pingtang recipe group were higher than those in normal control group 16 weeks after modeling （P 〈 0.01）, but there was no significant difference between model group and pingtang recipe group.⑨ Pathomorphological changes in liver：. There was no abnormality in normal control group 16 weeks after modeling. Diffuse fatty degeneration was observed in hepatocyte of rats in model group and pingtang recipe group, but changes of hepatic lipid was not different between two groups.
CONCLUSION： Pingtang recipe can improve blood glucose, lipid, resistance of insulin and liver function in rats with type 2 diabetes mellitus complicated with fatty liver, but cannot relieve the damage of hepatic histology.
出处
《中国临床康复》
CAS
CSCD
北大核心
2006年第11期77-80,共4页
Chinese Journal of Clinical Rehabilitation
基金
福建省卫生厅基金资助项目(2002Y05)~~
