糖肾颗粒对糖尿病肾病早期的干预及其机制的实验研究

发布时间：2017-12-28 08:22

本文关键词：糖肾颗粒对糖尿病肾病早期的干预及其机制的实验研究　出处：《湖北中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的通过单侧肾脏切除、高脂高糖饮食联合尾静脉注射STZ的方式将Wistar大鼠诱导为早期糖尿病肾病模型,观察糖肾颗粒对早期糖尿病肾病的防治作用,并探讨其可能的分子机制。方法SPF级雄性Wistar大鼠90只,适应性喂养1周后,随机分为正常组(A组)、模型组(B组)、糖肾颗粒低量组组(C组)、糖肾颗粒中剂量组(D组)、糖肾颗粒高量(E组)、贝那普利组(F组)各15只。除A组外,其余各组均将左肾摘除。术后1周,A组予常规饲料,其余各组予高脂高糖饲料。4周后,除A组外,各组大鼠按30mg/kg的计量尾静脉注射STZ,72小时后测空腹血糖和尿糖,若达到以下标准则认为DN模型成立:(1)空腹血糖≥16.7mmol/L;(2)尿糖阳性;(3)尿量大于正常组的50%。A,B组予蒸馏水灌胃,C组、D组、E组采用低、中、高剂量糖肾颗粒灌胃,F组采用盐酸贝那普利灌胃。各组均按10ml/kg剂量灌胃,治疗时间为6周。给药第6周末,留取尿液测微量白蛋白;禁食12h后,用1%戊巴比妥钠麻醉,腹主动脉取血,经离心留取血清,用于测定各项生化指标;采用PBS(PH7.4)灌注2min至肾脏变白后快速钳夹左肾称重,纵切为两部分,一部分保存于4%甲醛中,用于光镜检测;另外一部分保存于液氮中,用于检测mTOR/p70S6K信号通路中mTOR、p70S6K等蛋白的含量。结果1.一般情况与正常组大鼠比较,DN大鼠均有不同程度的饮食增多,饮水增多,尿量增多,形体消瘦,精神不振,活动迟缓。2.体重与相对肾重的情况与正常组大鼠比较,模型组大鼠体重明显下降,而相对肾重明显增加,差异具有统计学意义(P0.05);各治疗组(C组、D组、E组、F组)大鼠体重比模型组增加,相对肾重减轻,差异具有统计学意义(P0.05)。3.各项检测指标的比较与A组大鼠比较,B组大鼠血糖、血脂、尿白蛋白均升高,差异具有统计学意义(P0.05),血肌酐、血尿素氮无显著性差异(P0.05)。与B组相比,中药治疗组(C组、D组、E组)尿蛋白均有不同程度下降,差异具有统计意义(P0.05),血肌酐、血尿素氮无显著性差异(P0.05)。与B组相比,F组尿蛋白降低,差异具有统计学意义(P0.05),血糖、血脂有差异,但是无统计学意义(P0.05),血肌酐、血尿素氮无显著性差异(P0.05)。4.肾脏病理形态的改变DN大鼠的肾小球明显增大,系膜区增宽,着色加深,肾小管明显扩张。各治疗组上述表现有不同程度的缓解。5.mTOR/p70S6K通路中mTOR、p70S6K等蛋白的表达情况各组肾脏组织中mTOR、p70S6K总蛋白含量无显著性差异(P0.05);与A组比较,B组肾组织中p-mTOR、p-p70S6K含量明显增多,差异具有统计学意义(P0.05);与B组相比,各治疗组(C组、D组、E组、F组)肾组织中p-mTOR、p-p70S6K含量明显减少,差异具有统计学意义(P0.05)。结论1.糖肾颗粒可以减少DN大鼠早期尿白蛋白,降低血糖,调节血脂代谢。2.糖肾颗粒可以抑制DN大鼠mRTOR/p70S6K信号通路的活性,这可能是糖肾颗粒发挥治疗作用的基础。
[Abstract]:Objective to induce Wistar rats into an early diabetic nephropathy model by unilateral nephrectomy, high fat and high carbohydrate diet combined with tail vein injection of STZ, observe the preventive and therapeutic effects of Tang Shen Granule on early diabetic nephropathy, and explore its possible molecular mechanism. Methods 90 SPF male Wistar rats were randomly divided into normal group (group A), model group (group B), sugar kidney granule low volume group (C group), sugar kidney granule middle dose group (D group), sugar kidney granule high (E group), benazepril group (F group) after 15 weeks of adaptive feeding for 1 weeks. Except for group A, the left kidney was removed in all the other groups. 1 weeks after the operation, group A was given conventional feed, and the other groups were given high fat and high sugar diet. After 4 weeks, except group A, rats in each group according to the measurement of intravenous injection of STZ 30mg/kg, after 72 hours of fasting blood glucose and urine glucose, should meet the following criteria is that the DN model established: (1) fasting blood glucose was more than 16.7mmol/L; (2) positive urine; (3) the amount of urine is higher than the normal group of 50%. Group B, group C, group D and E group were fed with low, medium and high dose of sugar kidney granules in group C, group E and group F with benazepril hydrochloride gavage. All groups were given gastric perfusion at 10ml/kg dose, and the treatment time was 6 weeks. Treatment for sixth weeks, urine trace albumin; after fasting 12h, with 1% sodium pentobarbital anesthesia, abdominal aortic blood serum after centrifugation, for the determination of biochemical indicators; using PBS (PH7.4) 2min to white kidney perfusion after rapid clamping left renal weight, cut into two parts, a partially preserved in 4% formaldehyde, used for light detection; the other part preserved in liquid nitrogen, for measuring the content of mTOR and p70S6K in the mTOR/p70S6K signaling pathway protein. Results of the 1. general and normal rats, DN rats have different levels of diet increased, drinking water increased, urine volume increased, emaciation, listlessness, slow. Comparison of 2. body weight and relative kidney weight and normal rats, body weight of rats in model group decreased significantly, while the relative kidney weight increased significantly, the difference was statistically significant (P0.05); the treatment group (C group, D group, E group, F group) increased the body weight of rats than in model group, relative kidney weight, the difference was statistically significant (P0.05). 3., compared with the A group, the blood glucose, blood lipid and urinary albumin in the B group increased. The difference was statistically significant (P0.05), but there was no significant difference in serum creatinine and blood urea nitrogen (P0.05). Compared with group B, the urine protein of Chinese medicine treatment group (group C, group D and group E) decreased in varying degrees, the difference was statistically significant (P0.05), but there was no significant difference in serum creatinine and blood urea nitrogen (P0.05). Compared with group B, the urine protein of group F was decreased, the difference was statistically significant (P0.05), blood glucose and blood lipid were different, but there was no statistical significance (P0.05), there was no significant difference in serum creatinine and blood urea nitrogen (P0.05). 4. the pathological changes of the kidneys were changed in DN rats. The glomerulus was enlarged, the mesangial region widened, the coloring was deepened, and the renal tubule dilated obviously. The above performance in the treatment group was relieved in varying degrees. MTOR and p70S6K protein in 5.mTOR/p70S6K pathway expression in kidney tissue were mTOR, p70S6K total protein content had no significant difference (P0.05); compared with group A, renal B group p-mTOR, p-p70S6K content increased, the difference was statistically significant (P0.05); compared with B group, the treatment group (C group, D group, E group, F group) in renal tissue p-mTOR, p-p70S6K content decreased significantly, the difference was statistically significant (P0.05). Conclusion 1. sugar kidney granules can reduce the early urinary albumin, reduce the blood sugar and regulate the metabolism of blood lipid in DN rats. 2. sugar kidney granules can inhibit the activity of mRTOR/p70S6K signaling pathway in DN rats, which may be the basis for the therapeutic effect of sugar kidney granules.
【学位授予单位】：湖北中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R259

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