發(fā)布時(shí)間：2018-11-21 15:42

【摘要】：目的：觀察溫陽(yáng)活血利水法對(duì)阿霉素腎病大鼠腎小球足細(xì)胞的骨架蛋白CD2AP、 a-actinin-4、 synaptopodin、Arp3及cortactin表達(dá)的影響。探討溫陽(yáng)活血利水方如何通過(guò)Nephrin到足突骨架蛋白及接頭蛋白,以及信號(hào)轉(zhuǎn)導(dǎo)途徑治療兒童腎病綜合征的作用機(jī)制。 方法：SPF級(jí)SD雄性大鼠42只,體重115±15克,將其適應(yīng)性喂養(yǎng)十天,使其體重增至200±30克,按隨機(jī)分層分組的原則將其分為4組,其中中藥組12只、正常組6只、強(qiáng)的松組12只、模型組12只,除正常組外,其余各組均采用阿霉素(ADR)5.5mg/kg一次性尾靜脈注射,誘導(dǎo)一種類似人類微小病變型腎病綜合征的大鼠腎病模型,正常組尾靜脈一次性注射生理鹽水1毫升。注射ADR一周后開始治療,中藥組每日一次經(jīng)胃灌服溫陽(yáng)活血利水方顆粒劑混懸液3毫升,強(qiáng)的松組按12mg/kg·d,經(jīng)胃灌服強(qiáng)的松懸液3毫升(強(qiáng)的松劑量同前),正常組和模型組每日灌服3毫升生理鹽水,均持續(xù)4周。觀察實(shí)驗(yàn)大鼠的體重、二便、水腫、舌質(zhì)等一般情況；造模前、造模后第1周末及第5周末分別測(cè)大鼠24小時(shí)尿蛋白定量；實(shí)驗(yàn)第6周末處死大鼠取材。(1)麻醉后心臟取4m1血,分離血清,檢測(cè)總蛋白(TP)、總膽固醇(CHOL)、甘油三酯(TG)的水平。(2)采用免疫熒光染色測(cè)定各組大鼠腎小球足細(xì)胞中骨架蛋白CD2AP、 a-actinin-4、 synaptopodin、 Arp3及cortactin的表達(dá)水平。(3)取腎組織光鏡檢測(cè)組織形態(tài)學(xué)變化；電鏡下檢測(cè)超微結(jié)構(gòu)變化。 結(jié)果：(1)一般情況：正常組大鼠毛發(fā)潤(rùn)澤,活動(dòng)自如,精神狀態(tài)較好,食欲佳。其余各組大鼠注射阿霉素10～14天后大鼠出現(xiàn)體重不增,反應(yīng)遲鈍,活動(dòng)明顯減少,弓背蜷縮,聚堆,體毛稀疏,甚至片狀脫落,進(jìn)食量及飲水量明顯減少,時(shí)有便溏。注射阿霉素15天后大鼠開始出現(xiàn)四足水腫,足、尾紅紫,舌質(zhì)紫暗。治療期間,各治療組分別用中藥、強(qiáng)的松治療2周后大鼠反應(yīng)逐漸好轉(zhuǎn),進(jìn)食及飲水量較前增加,活動(dòng)增加；其中中藥組尿量明顯增多,體重緩慢回升。強(qiáng)的松組大鼠表現(xiàn)為躁動(dòng)不安,易激惹,攻擊性強(qiáng)。 (2)尿液及血液檢測(cè)結(jié)果：造模后1周,各造模組大鼠24小時(shí)尿蛋白定量與注射前比較有所升高,兩周后增加明顯。溫陽(yáng)活血利水方和強(qiáng)的松在治療4周后能顯著降低阿霉素腎病大鼠的尿蛋白水平,而中藥組和強(qiáng)的松組之間無(wú)明顯差異。阿霉素腎病大鼠存在明顯的低蛋白血癥和高脂血癥,溫陽(yáng)活血利水方和強(qiáng)的松都能在治療5周后升高阿霉素腎病大鼠的血清蛋白水平,但僅溫陽(yáng)活血利水方能降低其血清脂質(zhì)水平。 (3)免疫熒光染色：阿霉素腎病大鼠腎小球足細(xì)胞蛋白CD2AP、 synaptopodin、 Arp3及cortactin表達(dá)降低,而溫陽(yáng)活血利水方和強(qiáng)的松能在一定程度上抑制四種骨架蛋白表達(dá)的下降,而模型組及治療組a-actinin-4的表達(dá)均無(wú)改變,但其分布存在差異。 (4)病理：溫陽(yáng)活血利水方可以有效改善阿霉素腎病大鼠腎組織足細(xì)胞的足突融合。 結(jié)論：溫陽(yáng)活血利水方可以降低阿霉素腎病大鼠的蛋白尿；控制膽固醇、血脂；增加血清總蛋白；減輕其腎組織足細(xì)胞的足突融合；抑制阿霉素腎病大鼠腎組織足細(xì)胞蛋白CD2AP、synaptopodin、 Arp3及cortactin表達(dá)的下降；
[Abstract]:Objective: To observe the effect of Wenyang Huoxue water method on the expression of bone frame protein CD2AP, a-actin-4, synaphodin, Arp3 and cortin in adriamycin-induced nephropathy rats. Objective To study the mechanism of the effect of Wenyang Huoxue Lishui Recipe on the treatment of children's nephrotic syndrome through the treatment of the protein and its protein, and the signal transduction pathway in the treatment of children's nephrotic syndrome. Methods: 42 SD male rats, weighing 115 to 15 g, were fed for 10 days, the weight was increased to 200 to 30 g, and the body weight was divided into 4 groups according to the principle of random stratified grouping, 12 of the traditional Chinese medicine group, 6 in the normal group and 12 in the strong group, and 1 in the model group. 2. In addition to the normal group, the other groups were treated with adriamycin (ADR) of 5. 5 mg/ kg for one-time tail vein to induce a rat kidney disease model similar to the human micro-pathological type nephrotic syndrome, and the normal group tail vein was injected with normal saline at one time. One week after the injection of ADR, the treatment was started, and the traditional Chinese medicine group was given 3 ml of suspension liquid of Wenyang Huoxue Huoxue prescription granule once daily through the stomach, and the prednisone group was administered with 3 ml of strong suspension liquid (the same dose as before), the normal group and the model group with 3 ml of physiological saline in the normal group and the model group. Brine, continuous 4 weeks. The body weight, edema, tongue and other general conditions of the experimental rats were observed. The 24-hour urine protein was measured on the first weekend and the fifth weekend before the model, and the rats were sacrificed at the end of the first week. Materials are obtained. (1) After anesthesia, the heart takes 4m1 blood, separates serum, and detects total protein (TP), total cholesterol (CHOL), and triglyceride (TG). level. (2) The expression of bone frame protein CD2AP, a-actin-4, synaptopdin, Arp3 and cortin in the rat's glomerular foot cells was determined by immunofluorescence staining. level. (3) the morphological changes of the tissue were detected by the light microscope of the renal tissue; the ultrastructures were examined under the electron microscope. Change. Result: (1) General situation: the normal group of rats' hair is moist, the activity is free, the mental state is higher, Good appetite. The rest of the rats were injected with adriamycin for 10 to 14 days. The rats showed no increase in body weight, slow response, and reduced activity. After 15 days of injection of adriamycin, the rats began to have four-foot edema, foot, and red violet. and the urine volume of the traditional Chinese medicine group is obviously increased, and the volume of the traditional Chinese medicine group is obviously increased, The heavy and slow recovery of the rats. The rats in the high-pressure group showed restlessness and irritability. (2) The results of urine and blood test: 1 week after the model, the 24-hour urine protein of each model group was higher than that of the pre-injection. and the level of the urinary protein in the adriamycin-induced nephropathy rats can be significantly reduced after 4 weeks of treatment by the Wenyang Huoxue Lishui prescription and the prednisone, and the traditional Chinese medicine group and the prednisone group There was no significant difference between the groups. There were obvious hypoproteinemia and hyperlipoidemia in the rats with adriamycin nephrosis. The serum protein level of adriamycin-induced nephropathy rats could be increased after 5 weeks of treatment. (3) Immunofluorescence staining: the expression of CD2AP, synaptopdin, Arp3 and cortin in the glomerular podocyte protein of the adriamycin-induced nephropathy rats decreased, while the Wenyang Huoxue and the prednisone could inhibit the decrease of the expression of the four kinds of skeletal proteins to a certain extent, while the model group and the treatment group a-actin None of the in-4 expression Change, but there is a difference in its distribution. (4) Pathology: Wenyang Huoxue Lishui Prescription can effectively improve the adriamycin kidney Conclusion: Wenyang Huoxue Lishui prescription can reduce the albuminuria in the adriamycin-induced nephropathy rats, control the cholesterol and blood lipid, increase the serum total protein, reduce the apophysis fusion of the renal tissue foot cells, and inhibit the renal tissue foot cell protein CD2AP, synaptopdin, Ar in the adriamycin-induced nephropathy rats.
【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】：2012
【分類號(hào)】：R272

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