间充质干细胞治疗慢性间质性肾炎动物模型意义初探

发布时间：2018-05-28 19:20

本文选题：慢性间质性肾炎 + 间充质干细胞　；参考：《中国医科大学》2010年硕士论文

【摘要】： 目的通过成功制备腺嘌呤所致慢性间质性肾炎(CIN)的动物模型,将培养的间充质(MSCs)通过尾静脉的方法注射入大鼠体内,进一步评价干细胞评价间充质干细胞在治疗慢性间质性肾炎动物模型治疗效果。方法采用腺嘌呤(100mg／kg／d)持续灌胃的方法,连续4周,此后停药4周,检测部分大鼠的临床、生化指标,判断动物模型的情况,在制备模型的过程中,另取Wistar大鼠,成功剥离股骨,取出骨髓至细胞悬液后,于培养皿中分离培养,当细胞基本贴壁、铺满瓶底,且大部分细胞培养成梭形时进行传代,连续传3代后备用,此后采用免疫组化方法,鉴定培养细胞CD90、CD34的表达,确定模型、细胞培养均成功后,将剩余大鼠随机分为三组(A、B和C),A组(移植组)尾静脉注射培养好的间充质干细胞,B组(非移植组)注入等量生理盐水,C组(对照组)不予干预,每日观察大鼠的临床表现、体重变化,至观察4周后,将大鼠处死,检测大鼠处死前的临床表现、肾功情况以及处死后肾脏的病理变化。结果动物模型制备情况：腺嘌呤灌胃组大鼠表现出明显的体重下降、多饮多尿、毛发脱落、精神萎顿、抽搐甚至死亡,将上述症状自制成临床症状评分表格,同血清肌酐进行相关分析,发现两者呈现明显的相关性,判断临床症状评分表格成功,同对照组相关,腺嘌呤灌胃组无论从临床评分、生化以及病理评分同对照组相比均表现出统计学差异(P0.05),且病理表现以肾小管、间质损害为主,小球损害轻微,判断动物模型制作成功,间充质干细胞：可见细胞培养贴满瓶底,细胞变形成梭形,部分融合,抗体表达CA90+、CD34-,判断间充质干细胞培养成功。在进行间充质干细胞治疗过程中,与正常对照组相比,移植组、非移植组从临床表现、生化指标以及病理结果均有不同程度的恶化,三组相比存在统计学差异(P0.05),但当移植组与非移植组进行比较时,两组差异并不明显,未见统计学差异(P0.05)。结论腺嘌呤大鼠动物模型制备成功,间充质干细胞不能逆转慢性间质性肾炎的临床、病理变化,未能取得预期的试验效果,在慢性间质性肾炎治疗中作用有限。
[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.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R692.3;R-332

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