【摘要】 目的:观察发酵虫草菌粉对糖尿病肾病模型大鼠的治疗效果。方法:实验于2004-02/05在新乡医学院第一附属医院中心实验室完成。①选用纯系SD大鼠45只,雄性23只,雌性22只。随机将大鼠分为3组:正常对照组、模型组、发酵虫草菌粉治疗组,每组15只。模型组和发酵虫草菌粉组:于禁食24h后行四氧嘧啶125mg/kg的剂量左下腹腔一次性注入制成糖尿病大鼠模型。正常对照组:注入同等剂量的生理盐水。72h后,随机血糖≥16.7mmol/L者为糖尿病模型造模成功。发酵虫草菌粉治疗组:给予发酵虫草菌粉[商品名:百令胶囊,主要成分为发酵虫草菌粉,杭州中美华东制药有限公司生产,批准文号:(91)卫药准字Z-81号,0.2g/粒]0.2g/(kg·d)灌胃,1次/d,共12周。模型组和正常对照组:灌入同等量的生理盐水,1次/d,共12周。②12周后,采用BACKMAN全自动生化分析仪测定血糖、血脂、肾功能、24h尿蛋白排泄量。测定双侧肾脏质量,计算肾重/体质量。采用Paisey方法测定组织中蛋白糖基化产物。采用光镜检查,HPIAS-1000型图像分析系统对病理切片进行分析,检测肾小球平均截面积,计算肾小球硬化指数(采用Raij等的半定量方法估计肾小球硬化程度,每张切片均观察20个肾小球,根据肾小球硬化灶占肾小球比例分为0～4分:0分,肾小球无病变;1分,受损范围≤25%;2分,受损范围26%50%;3分,受损范围51%～75%;4分受损范围≥76%)。电镜观察肾小球滤过屏障情况,测定肾小球基底膜平均厚度和平均肾小球截面积,并观察肾小管病变情况。③组间比较采用方差分析。结果:大鼠45只均进入结果分析。①模型组大鼠空腹血糖、三酰甘油和总胆固醇水平,肾质量/体质量,24h尿蛋白排泄量,肾脏组织中蛋白糖基化产物含量明显高于正常对照组(P<0.05～0.01)。发酵虫草菌粉治疗组大鼠症状明显较模型组减轻,空腹血糖、三酰甘油和总胆固醇水平,肾质量/体质量,24h尿蛋白排泄量,肾脏组织中蛋白糖基化产物含量低于模型组(P<0.05～0.01)。各组大鼠血尿素氮及血肌酐水平均在正常范围,组间比较,差异不明显(P>0.05)。②光镜检查结果显示,模型组大鼠肾小球截面积明显大于正常对照组(P<0.01),系膜区基质增生扩大,发酵虫草菌粉治疗组明显小于模型组(P<0.01);模型组大鼠肾小球硬化明显,平均肾小球硬化指数明显高于正常对照组(P<0.01),发酵虫草菌粉治疗组明显小于模型组(P<0.01)。③电镜结果显示,模型组大鼠肾小球基底膜显著的不均匀增厚,明显高于正常对照组(P<0.01),并伴有广泛的肾小管上皮细胞的髓样变性及胞浆空泡化变性。发酵虫草菌粉治疗组上述病变明显减轻(P<0.05～0.01)。结论:发酵虫草菌粉能显著改善糖尿病大鼠肾脏结构,防止系膜区扩大和基底膜增厚,具有显著地抗肾小球纤维化和防止肾小管病变的作用。更多还原

【Abstract】 AIM: To observe therapeutic effects of zymoyic fungal powder of cordyceps sinensis on diabetic nephropathy (DN) in rats. METHODS: The experiment was completed in the Central Laboratory of First Hospital Affiliated to Xinxiang Medical College from February to May 2004. ① Forty-five SD rats (23 males and 22 females) were divided randomly into 3 groups: normal control group, model group and drug group with 15 in each group. The diabetic models were established by intraperitoneal injection of 125 mg/kg alloxan in model group and drug group 24 hours after fasting. Rats in normal control group were injected with the same volume of saline. If volume of random blood glucose was not less than 16.7 mmol/L after 72 hours, the diabetic models were successful. Rats in drug group were treated with 0.2 g/(kg·d) zymoyic fungal powder of cordyceps sinensis [bailing capsule: main components of zymoyic fungal powder of cordyceps sinensis, provided by Hangzhou Chinese-American Huadong Pharmaceutical Company Limited, batch number: (91) wei yao zhun zi Z-81, 0.2 g/pill] once a day for 12 weeks. Rats in model group and normal control group were perfused with the same volume of saline once a day for 12 weeks.② After 12 weeks, levels of blood glucose, lipids, renal function and excretion of 24-hour urine protein were assayed with BACKMAN automatic biochemistry analyzer. The weight of both kidneys and the kidney weight/body mass were measured. Glucosylation of protein (GP) was assayed with Paisey method; pathological sections were analyzed with light microscope and HPIAS-1000 imaging analysis system; average section area of renal corpuscle was measured and index of glomerulosclerosis was calculated. Levels of glomerulosclerosis were evaluated with semi-quantitative method by Raij et al. Twenty renal corpuscles were observed in each section. According to ratio of renal corpuscle accounted for glomerulosclerosis focus, scores were classified 5 grades: 0 point: renal corpuscle was not in lesion; 1 point: damaged area ≤25%; 2 points: damaged area from 26% to 50%; 3 points: damaged area from 51% to 75%; 4 points: damaged area ≥76%. Renal corpuscle filtrating barrier was observed by electron microscope to measure average depth of glomerular basement membrane and average area of glomerular plana and to observe lesions of renal corpuscle. ③ Differences among groups were compared with analysis of variance. RESULTS: Totally 45 rats entered the final analysis. ① Contents of fasting blood glucose, triacylglycerol (TG) and total cholesterol (TC), ration between renal mass and body mass, excretion of 24-hour urine protein, and content of GP in renal tissue of rats in model group were higher than those in normal control group (P < 0.05-0.01). Symptoms in drug group were relieved as compared with those in model group, and contents of fasting blood glucose, TG and TC, ration between renal mass and body mass, excretion of 24-hour urine protein, and content of GP in renal tissue were lower than those in model group (P < 0.05-0.01). Contents of blood urea nitrogen and blood creatinine were both normal, and there were not significantly different among groups (P > 0.05). ② Results under light microscope showed that area of glomerular plana in model group was larger than that in normal control group (P < 0.01). Proliferation of glomerular basement membrane was extended and that was smaller in drug group than that in model group (P < 0.01). Glomerulosclerosis was obvious in model group, and average index of glomerulosclerosis was higher than that in normal control group (P < 0.01), and that in drug group was lower than that in model group (P < 0.01). ③ Results under electron microscope showed that glomerular basement membrane in model group was uneven thick significantly, and there was extensive vacuolar and medullary degeneration of epithelial cells in renal tubules. The thickness in model group was higher than that in normal control group (P < 0.01). Those abnormal changes were alleviated significantly in drug group (P < 0.05-0.01). CONCLUSION: Zymoyic fungal powder of cordyceps sinensis can alleviate the renal structure in diabetic rats, prevent enlargement of glomerular plana area and thickening of basement membrane, has some effects on anti-glomerulosclerosis and prevention of renal tubular lesion.更多还原