本文選題：慢性間質(zhì)性腎炎 + 間充質(zhì)干細胞��； 參考：《中國醫(yī)科大學》2010年碩士論文

【摘要】： 目的 通過成功制備腺嘌呤所致慢性間質(zhì)性腎炎(CIN)的動物模型,將培養(yǎng)的間充質(zhì)(MSCs)通過尾靜脈的方法注射入大鼠體內(nèi),進一步評價干細胞評價間充質(zhì)干細胞在治療慢性間質(zhì)性腎炎動物模型治療效果。 方法 采用腺嘌呤(100mg／kg／d)持續(xù)灌胃的方法,連續(xù)4周,此后停藥4周,檢測部分大鼠的臨床、生化指標,判斷動物模型的情況,在制備模型的過程中,另取Wistar大鼠,成功剝離股骨,取出骨髓至細胞懸液后,于培養(yǎng)皿中分離培養(yǎng),當細胞基本貼壁、鋪滿瓶底,且大部分細胞培養(yǎng)成梭形時進行傳代,連續(xù)傳3代后備用,此后采用免疫組化方法,鑒定培養(yǎng)細胞CD90、CD34的表達,確定模型、細胞培養(yǎng)均成功后,將剩余大鼠隨機分為三組(A、B和C),A組(移植組)尾靜脈注射培養(yǎng)好的間充質(zhì)干細胞,B組(非移植組)注入等量生理鹽水,C組(對照組)不予干預(yù),每日觀察大鼠的臨床表現(xiàn)、體重變化,至觀察4周后,將大鼠處死,檢測大鼠處死前的臨床表現(xiàn)、腎功情況以及處死后腎臟的病理變化。 結(jié)果 動物模型制備情況：腺嘌呤灌胃組大鼠表現(xiàn)出明顯的體重下降、多飲多尿、毛發(fā)脫落、精神萎頓、抽搐甚至死亡,將上述癥狀自制成臨床癥狀評分表格,同血清肌酐進行相關(guān)分析,發(fā)現(xiàn)兩者呈現(xiàn)明顯的相關(guān)性,判斷臨床癥狀評分表格成功,同對照組相關(guān),腺嘌呤灌胃組無論從臨床評分、生化以及病理評分同對照組相比均表現(xiàn)出統(tǒng)計學差異(P0.05),且病理表現(xiàn)以腎小管、間質(zhì)損害為主,小球損害輕微,判斷動物模型制作成功,間充質(zhì)干細胞：可見細胞培養(yǎng)貼滿瓶底,細胞變形成梭形,部分融合,抗體表達CA90+、CD34-,判斷間充質(zhì)干細胞培養(yǎng)成功。在進行間充質(zhì)干細胞治療過程中,與正常對照組相比,移植組、非移植組從臨床表現(xiàn)、生化指標以及病理結(jié)果均有不同程度的惡化,三組相比存在統(tǒng)計學差異(P0.05),但當移植組與非移植組進行比較時,兩組差異并不明顯,未見統(tǒng)計學差異(P0.05)。 結(jié)論 腺嘌呤大鼠動物模型制備成功,間充質(zhì)干細胞不能逆轉(zhuǎn)慢性間質(zhì)性腎炎的臨床、病理變化,未能取得預(yù)期的試驗效果,在慢性間質(zhì)性腎炎治療中作用有限。
[Abstract]:Purpose The rat model of chronic interstitial glomerulonephritis induced by adenine was successfully established, and the cultured mesenchymal cells (MSCs) were injected into the rat via tail vein. To evaluate the efficacy of mesenchymal stem cells in the treatment of chronic interstitial glomerulonephritis. Method After 4 weeks of continuous administration of adenine (100 mg / kg / d), the clinical and biochemical indexes of some rats were detected, and the animal model was judged. During the course of making the model, the femur was removed successfully from the Wistar rats. After the bone marrow was removed into the cell suspension, the cells were isolated and cultured in a culture dish. When the cells were basically adhered to the wall, covered with the bottom of the bottle, and most of the cells were cultured into fusiform shape, the cells were passed on for 3 consecutive generations, and then the cells were prepared by immunohistochemical method. The expression of CD90 and CD34 in cultured cells was identified, and the model was established. The remaining rats were randomly divided into three groups: caudal vein injection of cultured mesenchymal stem cells group B (non-transplantation group) and control group (control group). The clinical manifestations of the rats were observed daily. After 4 weeks of observation, the rats were killed and the clinical manifestations, renal function and pathological changes of the kidneys were measured before and after the rats were killed. Result Animal model preparation: rats in adenine group showed significant weight loss, polyuria, hair loss, mental collapse, convulsion and even death. Correlation analysis with serum creatinine showed that there was a significant correlation between them. The clinical symptom score table was successful. Compared with the control group, the biochemical and pathological scores showed statistical difference (P 0.05), and the pathological findings were mainly renal tubule and interstitial damage, and the lesion of the pellets was slight. The mesenchymal stem cells were observed to be covered with the bottoms of the bottle, and the mesenchymal stem cells were found to be successful in making the animal model. The cells were transformed into fusiform, partially fused, and the antibody expressed CA90 CD34. It was judged that the mesenchymal stem cells were cultured successfully. In the course of mesenchymal stem cell therapy, compared with the normal control group, the clinical manifestations, biochemical indexes and pathological results of the transplantation group and the non-transplantation group all deteriorated in varying degrees. There was a statistical difference between the three groups (P 0.05), but there was no significant difference between the two groups when compared with the non-transplantation group (P 0.05). Conclusion Adenine rat model was successfully established, mesenchymal stem cells could not reverse the clinical and pathological changes of chronic interstitial glomerulonephritis, failed to obtain the expected experimental results, and played a limited role in the treatment of chronic interstitial nephritis.
【學位授予單位】：中國醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R692.3;R-332

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