Zhang D W, Qiu H, Mei Y M, Fu H, Zheng H G
Department of Nephrology, General Hospital of Shenyang Military Area, Shenyang, China.
J Biol Regul Homeost Agents. 2015 Jul-Sep;29(3):609-17.
This study aimed to explore the influence of umbilical cord mesenchymal stem cells (UMSC) on stem cell homing and glomerular mesangial cell (GMC) after intravenous injection performed on mice tails with IgA nephropathy (IgAN) and its possible mechanism, which provide a new way and theoretical basis for the application of stem cell transplantation (SCT) in kidney disease treatment. Specific pathogen free (SPF) male Kunming mice were randomly divided into groups. A complex method applying bovine serum albumin (BSA) gavage, hypodermic injection of CCl4 and lipopolysaccharide (LPS) was used for building IgAN mice model. In addition, vascular endothelial growth factor (VEGF), connective tissue growth factor (CTGF) and cluster of differentiation (CD) 44 were observed by Masson staining and detected with immunohistochemistry (IHC) to confirm homing and location of mesenchymal stem cells (MSCs). Moreover, Western Blot was used for detecting VEGF and CTGF so as to explore the possible mechanism of applying UMSC in treating IgAN. Masson staining indicated that fibrosis degree of MSCs in treatment group was significantly lower than in negative control group after stem cell treatment. Routine urine test explained that proteinuria in treatment group were (7.15±0.31), (4.87±0.22), (2.95±0.16) g/24 h and (12.00±1.38) g/24 h in model group (P less than 0.05). MSCs were observed to be located in glomerulus and renal interstitium by IHC detection of CD44 and IHC qualitative observation of VEGF and CTGF had different positive expressions in three groups. Furthermore, different expressions of VEGF and CTGF were observed quantitatively by Western Blot. Fibrosis degree of renal tissue relieves, hematuresis and proteinuria eases and IgAN symptoms obviously improve after UMSC treatment, which hints that the treatment of HUMSC has protective effect on IgAN mice model.
本研究旨在探讨脐带间充质干细胞(UMSC)对尾静脉注射免疫球蛋白A肾病(IgAN)小鼠后干细胞归巢及肾小球系膜细胞(GMC)的影响及其可能机制,为干细胞移植(SCT)在肾脏疾病治疗中的应用提供新途径和理论依据。将无特定病原体(SPF)雄性昆明小鼠随机分组。采用牛血清白蛋白(BSA)灌胃、皮下注射四氯化碳(CCl4)和脂多糖(LPS)的复合方法构建IgAN小鼠模型。此外,通过Masson染色观察血管内皮生长因子(VEGF)、结缔组织生长因子(CTGF)和分化簇(CD)44,并采用免疫组织化学(IHC)检测以确认间充质干细胞(MSC)的归巢和定位。此外,采用蛋白质免疫印迹法(Western Blot)检测VEGF和CTGF,以探讨UMSC治疗IgAN的可能机制。Masson染色显示干细胞治疗后治疗组MSC的纤维化程度明显低于阴性对照组。尿常规检查显示治疗组蛋白尿分别为(7.15±0.31)、(4.87±0.22)、(2.95±0.16)g/24 h,模型组为(12.00±1.38)g/24 h(P<0.05)。通过CD44免疫组化检测观察到MSC位于肾小球和肾间质,VEGF和CTGF的免疫组化定性观察在三组中有不同的阳性表达。此外,通过蛋白质免疫印迹法(Western Blot)定量观察到VEGF和CTGF的不同表达。UMSC治疗后肾组织纤维化程度减轻,血尿和蛋白尿缓解,IgAN症状明显改善,提示人脐带间充质干细胞(HUMSC)治疗对IgAN小鼠模型有保护作用。