本文關(guān)鍵詞：糖腎顆粒對(duì)糖尿病腎病早期的干預(yù)及其機(jī)制的實(shí)驗(yàn)研究　出處：《湖北中醫(yī)藥大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 糖腎顆粒 糖尿病腎病 mRTOR/p70S6K信號(hào)通路

【摘要】：目的通過單側(cè)腎臟切除、高脂高糖飲食聯(lián)合尾靜脈注射STZ的方式將Wistar大鼠誘導(dǎo)為早期糖尿病腎病模型,觀察糖腎顆粒對(duì)早期糖尿病腎病的防治作用,并探討其可能的分子機(jī)制。方法SPF級(jí)雄性Wistar大鼠90只,適應(yīng)性喂養(yǎng)1周后,隨機(jī)分為正常組(A組)、模型組(B組)、糖腎顆粒低量組組(C組)、糖腎顆粒中劑量組(D組)、糖腎顆粒高量(E組)、貝那普利組(F組)各15只。除A組外,其余各組均將左腎摘除。術(shù)后1周,A組予常規(guī)飼料,其余各組予高脂高糖飼料。4周后,除A組外,各組大鼠按30mg/kg的計(jì)量尾靜脈注射STZ,72小時(shí)后測(cè)空腹血糖和尿糖,若達(dá)到以下標(biāo)準(zhǔn)則認(rèn)為DN模型成立:(1)空腹血糖≥16.7mmol/L;(2)尿糖陽(yáng)性;(3)尿量大于正常組的50%。A,B組予蒸餾水灌胃,C組、D組、E組采用低、中、高劑量糖腎顆粒灌胃,F組采用鹽酸貝那普利灌胃。各組均按10ml/kg劑量灌胃,治療時(shí)間為6周。給藥第6周末,留取尿液測(cè)微量白蛋白;禁食12h后,用1%戊巴比妥鈉麻醉,腹主動(dòng)脈取血,經(jīng)離心留取血清,用于測(cè)定各項(xiàng)生化指標(biāo);采用PBS(PH7.4)灌注2min至腎臟變白后快速鉗夾左腎稱重,縱切為兩部分,一部分保存于4%甲醛中,用于光鏡檢測(cè);另外一部分保存于液氮中,用于檢測(cè)mTOR/p70S6K信號(hào)通路中mTOR、p70S6K等蛋白的含量。結(jié)果1.一般情況與正常組大鼠比較,DN大鼠均有不同程度的飲食增多,飲水增多,尿量增多,形體消瘦,精神不振,活動(dòng)遲緩。2.體重與相對(duì)腎重的情況與正常組大鼠比較,模型組大鼠體重明顯下降,而相對(duì)腎重明顯增加,差異具有統(tǒng)計(jì)學(xué)意義(P0.05);各治療組(C組、D組、E組、F組)大鼠體重比模型組增加,相對(duì)腎重減輕,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。3.各項(xiàng)檢測(cè)指標(biāo)的比較與A組大鼠比較,B組大鼠血糖、血脂、尿白蛋白均升高,差異具有統(tǒng)計(jì)學(xué)意義(P0.05),血肌酐、血尿素氮無顯著性差異(P0.05)。與B組相比,中藥治療組(C組、D組、E組)尿蛋白均有不同程度下降,差異具有統(tǒng)計(jì)意義(P0.05),血肌酐、血尿素氮無顯著性差異(P0.05)。與B組相比,F組尿蛋白降低,差異具有統(tǒng)計(jì)學(xué)意義(P0.05),血糖、血脂有差異,但是無統(tǒng)計(jì)學(xué)意義(P0.05),血肌酐、血尿素氮無顯著性差異(P0.05)。4.腎臟病理形態(tài)的改變DN大鼠的腎小球明顯增大,系膜區(qū)增寬,著色加深,腎小管明顯擴(kuò)張。各治療組上述表現(xiàn)有不同程度的緩解。5.mTOR/p70S6K通路中mTOR、p70S6K等蛋白的表達(dá)情況各組腎臟組織中mTOR、p70S6K總蛋白含量無顯著性差異(P0.05);與A組比較,B組腎組織中p-mTOR、p-p70S6K含量明顯增多,差異具有統(tǒng)計(jì)學(xué)意義(P0.05);與B組相比,各治療組(C組、D組、E組、F組)腎組織中p-mTOR、p-p70S6K含量明顯減少,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論1.糖腎顆�？梢詼p少DN大鼠早期尿白蛋白,降低血糖,調(diào)節(jié)血脂代謝。2.糖腎顆�？梢砸种艱N大鼠mRTOR/p70S6K信號(hào)通路的活性,這可能是糖腎顆粒發(fā)揮治療作用的基礎(chǔ)。
[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.
【學(xué)位授予單位】：湖北中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R259

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