本文選題：阿霉素模型 + 局灶節(jié)段性腎小球硬化��； 參考：《北京中醫(yī)藥大學》2017年碩士論文

【摘要】：目的:通過觀察益腎活血方對FSGS大鼠模型足細胞損傷的保護作用,探討其機制研究。方法:采用隨機對照的實驗方法,實驗動物選用SD大鼠兩次尾靜脈注射阿霉素(4mg/kg/次)的方法制備的FSGS大鼠模型。二次尾靜脈注射間隔2周,于第二次尾靜脈注射后7、14天測24小時尿蛋白定量,大于100mg/24h提示造模成功,將提示造模成功的大鼠常規(guī)飼養(yǎng)至第二次尾靜脈注射后8周末,檢測24小時尿蛋白定量。剔除造模失敗、尾部潰爛嚴重或死亡大鼠,造模組剩余35只,計入實驗。根據(jù)大鼠第二次尾靜脈注射后第8周末24小時尿蛋白定量、體重,隨機分為:模型組(9只)、貝那普利組(8只)、益腎活血方低劑量組(9只)、益腎活血方高劑量組(9只);另設10只同周齡正常SD大鼠為正常組;各組連續(xù)干預8周。實驗過程中觀察記錄大鼠一般情況、體重、24小時尿蛋白定量水平;治療前眼眶取血檢測血漿白蛋白(ALB)、血肌酐(Scr)、尿素氮(BUN)水平;治療8周后大鼠麻醉、腹主動脈取血,分別凍存和固定腎臟,檢測血清中ALB、Scr、BUN水平;行HE、PAS染色觀察腎臟病理形態(tài)變化,檢測腎小球硬化程度;行Western blotting檢測WT1(Wilms' tumor 1)蛋白表達;行逆轉(zhuǎn)錄-多聚酶鏈反應(RT-PCR)檢測大鼠腎臟podocin、nephrinmRNA表達。所有數(shù)據(jù)采用SPSS 20.0統(tǒng)計學軟件進行分析處理。結(jié)果:①益腎活血方能夠明顯改善大鼠的一般生存狀態(tài)。②體重上,成模后,正常組與造模組之間即有統(tǒng)計學差異(P0.01);藥物干預后,高劑量組與貝那普利組體重仍有增加,模型組自治療后4周(實驗第12周)開始有下降趨勢,治療8周高劑量組、貝那普利組與模型組之間有統(tǒng)計學差異(P0.05)。③尿蛋白上,益腎活血方治療組、貝那普利組與模型組之間有統(tǒng)計學差異(P0.05);各治療組之間沒有差異。④血漿白蛋白上,治療后高劑量組與貝那普利組均較前有改善,與正常組之間沒有統(tǒng)計學差異(P0.05)。⑤腎功能上,血肌酐:高劑量組改善最為明顯,與模型組有明顯統(tǒng)計學差異(P0.01),并且與低劑量組與統(tǒng)計學差異(P0.05);尿素氮:貝那普利組改善最為明顯,與模型組相比,高劑量、貝那普利組有統(tǒng)計學差異(P0.05)。⑥組織病理學上,腎小球硬化指數(shù)、基質(zhì)比值治療后均有改善,雖然與正常組相比仍有明顯統(tǒng)計學差異(P0.01),但較模型組有統(tǒng)計學改善(P0.05)。⑦蛋白及基因表達上,治療后,益腎活血方治療組、貝那普利組WT1表達較模型組增加,有統(tǒng)計學差異(P0.05);益腎活血方高劑量組、貝那普利組nephrin和podocinmRNA水平與模型組相比有明顯統(tǒng)計學差異(P0.01)。結(jié)論:益腎活血方具有改善FSGS大鼠低蛋白血癥、蛋白尿和腎功能,減輕腎小球硬化程度的作用,其機制可能是增加nephrin、podocin基因表達和WT1蛋白表達。
[Abstract]:Aim: to investigate the protective effect of Yishen Huoxue recipe on podocyte injury in FSGS rats. Methods: the FSGS rat model was established by injecting adriamycin 4 mg / kg twice into the tail vein of SD rats. Two weeks after the second caudal vein injection, 24 hours urine protein was measured at 714 days after the second caudal vein injection, which was larger than that indicated by 100mg/24h. The rats who indicated the success of the second caudal vein injection were fed regularly until 8 weeks after the second caudal vein injection. Urine protein was measured in 24 hours. The remaining 35 rats in the model group were included in the experiment. According to the 24 hour urine protein quantity at the end of the eighth week after the second caudal vein injection, The rats were randomly divided into three groups: model group (n = 9), benazepril group (n = 8), low dose group (n = 9), high dose group (n = 9) and control group (n = 10). During the course of the experiment, we observed and recorded the general situation of rats, the weight of rats was measured by 24 hours urine protein; before treatment, the blood samples were taken from orbit to detect the levels of plasma albumin, creatinine, bun and bun; after 8 weeks of treatment, the rats were anesthetized, and the abdominal aorta was taken to get blood. Frozen and fixed kidneys were used to detect the level of serum ALB Scrn bun, the histopathologic changes of kidney and glomerulosclerosis were observed by HEPAS staining, and WT1 Wilms' tumor 1 protein expression was detected by Western blotting. The expression of podocinine mRNA in rat kidney was detected by reverse transcription-polymerase chain reaction (RT PCR). All the data were analyzed by SPSS 20.0 statistical software. Results the general living state of rats was significantly improved by using the "1: 1 Yishen Huoxue recipe". After the model was established, there was a significant difference between the normal group and the model group (P 0.01), and after the intervention of the drug, the weight of the high dose group and benazepril group still increased. In the model group, there was a downward trend since 4 weeks after treatment (the 12th week of the experiment). After 8 weeks of treatment, there was a significant difference between the benazepril group and the model group (P 0.05.3 urine protein), and the treatment group with Yishen Huoxue recipe. There was statistical difference between benazepril group and model group (P 0.05), there was no difference of 4. 4 plasma albumin between each treatment group, after treatment, there was improvement in high dose group and benazepril group, and there was no significant difference in renal function between benazepril group and normal group. Serum creatinine: the improvement was most obvious in the high dose group, which was significantly different from the model group (P 0.01), and was significantly different from the low dose group and the statistical difference (P 0.05), the urea nitrogen: benazepril group was the most obvious improvement, compared with the model group, the high dose was higher than that in the model group. In benazepril group, the histopathology of benazepril was significantly different. Glomerulosclerosis index and matrix ratio were all improved after treatment. Compared with the normal group, the expression of WT1 in the Yishen Huoxue prescription group and benazepril group was increased after treatment, but the protein and gene expression of P0.05.7 protein were significantly improved compared with the model group, although there was still significant difference between the treatment group and the normal control group, the expression of WT1 in the benazepril group was significantly higher than that in the model group. The levels of nephrin and podocinmRNA in the high dose group and benazepril group were significantly different from those in the model group. Conclusion: Yishen Huoxue recipe can improve hypoproteinemia, proteinuria and renal function and alleviate glomerular sclerosis in FSGS rats. Its mechanism may be to increase the expression of nephrininpodocin gene and WT1 protein.
【學位授予單位】：北京中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R285.5

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