人脐带间充质干细胞对IgA肾病模型小鼠的保护作用研究

发布时间：2018-05-31 00:05

本文选题：IgA肾病 + 小鼠　；参考：《辽宁医学院》2012年硕士论文

【摘要】：目的探讨脐带间充质干细胞对IgA肾病小鼠模型尾静脉注射后干细胞的归巢及对肾小球系膜细胞的影响及其可能机制，，为干细胞移植在肾脏病治疗领域中的应用提供新的途径及理论依据。方法 SPF级雄性昆明小鼠60只，按随机数字法随机分为对照组、模型组和人脐带间充质干细胞治疗组（以下简称治疗组）。利用牛血清白蛋白灌胃、皮下注射CCl4，应用脂多糖的复合方法造成小鼠IgA肾病模型。14周造模成功，尾静脉注射人脐带间充质干细胞，于第7、21、56天处死小鼠取材，做免疫荧光染色证实造模成功，Masson染色观察肾组织纤维化情况，免疫组化检测VEGF、CTGF，免疫组化检测CD44确定间充质干细胞归巢定位情况。Western Blot检测VEGF、CTGF蛋白表达量，探讨应用人脐带间充质干细胞治疗IgA肾病的可能机制。结果造模成功后，模型组和治疗组小鼠均出现蛋白尿，25只小鼠出现镜下血尿，15只小鼠肉眼血尿，其中模型组13只小鼠出现镜下血尿，7只小鼠出现肉眼血尿，治疗组12只小鼠出现镜下血尿，8只小鼠出现肉眼血尿；免疫荧光显示IgA沉积于肾小球系膜区；干细胞治疗后，MASSON染色显示人间充质干细胞治疗组纤维化程度明显较阴性对照组减轻，尿常规：24h蛋白尿治疗组（7.15±0.31）、（4.87±0.22）、（2.95±0.16）g/24h，模型组（12.00±1.38）g/24h,均有P0.05。免疫组化检测CD44观察间充质干细胞定位于肾小球及肾间质内，免疫组化定性观察VEGF、CTGF在三组中分别有不同的阳性表达，Western Blot定量观察VEGF、CTGF蛋白在三组中有不同的表达。结论在应用脐带间充质干细胞后，肾组织纤维化程度减轻，血尿蛋白尿减轻，IgA肾病症状明显改善，提示人脐带间充质干细胞治疗对IgA肾病小鼠模型具有保护作用。人脐带间充质干细胞注入IgA肾病小鼠模型后，可以通过趋化作用归巢到肾组织内，通过分泌细胞因子营造出适合微环境，使肾祖/干细胞重新分化为肾组织细胞，并通过调控VEGF、CTGF的表达水平来促使新生血管生成，减轻肾组织纤维化，修复受损肾小球系膜组织，使IgA肾病的一系列症状得到改善，从而达到治疗IgA肾病的目的。
[Abstract]:Purpose To investigate the effect of umbilical cord mesenchymal stem cells on homing of stem cells and on glomerular Mesangial cells after caudal vein injection in mice with IgA nephropathy. To provide a new approach and theoretical basis for stem cell transplantation in the field of kidney disease treatment. Method Sixty male Kunming mice of SPF grade were randomly divided into control group, model group and human umbilical cord mesenchymal stem cell treatment group (hereinafter referred to as the treatment group). Bovine serum albumin (BSA) was perfused into the stomach and CCL4 was injected subcutaneously. The IgA nephropathy model of mice was established successfully by using lipopolysaccharide compound method. Human umbilical cord mesenchymal stem cells were injected into caudal vein, and the mice were killed on day 7, 21 and 56. The renal fibrosis was observed by Masson staining, the expression of VEGF CTGF was detected by immunohistochemistry, the homing location of mesenchymal stem cells was determined by immunohistochemistry, and the expression of VEGF CTGF protein was detected by Western Blot. To explore the possible mechanism of human umbilical cord mesenchymal stem cells in the treatment of IgA nephropathy. Result After the establishment of the model, the proteinuria was found in 25 mice in both the model group and the treatment group, and in 15 mice with hematuria under microscope, among them, 13 mice in the model group showed hematuria under microscope and 7 mice showed hematuria with naked eye. In the treatment group, 12 mice appeared hematuria under microscope, 8 mice showed naked hematuria, and IgA was deposited in glomerular Mesangial area by immunofluorescence. After the treatment of stem cells, the degree of fibrosis in the human mesenchymal stem cells group was significantly less than that in the negative control group. The urine routine 24 hours proteinuria group (7.15 卤0.31) had 2.95 卤0.16 g / 24 h, and the model group was 12.00 卤1.38 g / r for 24 h, all with P0.05. CD44 was used to detect the location of mesenchymal stem cells (MSCs) in glomeruli and renal interstitium. The positive expression of VEGF CTGF in the three groups was observed qualitatively. Western Blot was used to quantitatively observe the different expression of VEGF CTGF protein in the three groups. Conclusion After the application of umbilical cord mesenchymal stem cells, the degree of renal fibrosis was alleviated, and the symptoms of IgA nephropathy were alleviated by hematuria proteinuria, suggesting that human umbilical cord mesenchymal stem cell therapy has protective effect on IgA nephropathy mice. After human umbilical cord mesenchymal stem cells were injected into IgA nephropathy mice model, they could homing into kidney tissue through chemotactic action, creating a suitable microenvironment by secreting cytokines, and redifferentiation of renal progenitor / stem cells into renal tissue cells. Through regulating the expression level of VEGF CTGF to promote neovascularization, alleviate renal fibrosis, repair the damaged glomerular Mesangial tissue, improve a series of symptoms of IgA nephropathy, so as to achieve the purpose of treating IgA nephropathy.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R692.3;R-332

【参考文献】