脂肪间充质干细胞对硬皮病小鼠皮损组织成纤维细胞增殖的影响及其机制  

Effects of adipose-derived mesenchymal stem cells on fibroblast proliferation in mouse scleroderma lesions and underlying mechanisms

作　　者：王维[1] 黄玉成[1] 陈燕辉[1] Wang Wei;Huang Yu-cheng;Chen Yan-hui(Zhengzhou People’s Hospital,Zhengzhou 450003,Henan Province,China)

机构地区：[1]郑州人民医院,河南省郑州市450003

出　　处：《中国组织工程研究》2018年第33期5249-5254,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:脂肪间充质干细胞在体外长期培养过程中始终保持多向分化潜能,可通过分泌多种抗纤维化细胞因子或生长因子,参与调节成纤维细胞增殖、抑制胶原沉积而发挥抗纤维化作用。目的:观察脂肪间充质干细胞对硬皮病小鼠皮损组织成纤维细胞增殖、凋亡的影响,并初步探讨相关机制。方法:从硬皮病小鼠模型皮损组织分离、纯化出成纤维细胞,经传代培养后,根据处理方式分为5组:空白对照组(A组)、脂肪间充质干细胞干预组(B组)、Wnt抑制剂HY-15597处理组(C组)、Wnt激活剂SB-216763处理组(D组)、脂肪间充质干细胞+SB-216763处理组(E组)。MTT法和流式细胞仪检测各组成纤维细胞增殖率和凋亡率;蛋白免疫印迹法检测各组成纤维细胞中Ⅰ型胶原、Ⅲ型胶原和Wnt信号通路相关蛋白表达水平。结果与结论:(1)与A组相比,B组和C组成纤维细胞增殖率均显著降低、凋亡率显著升高,D组增殖率显著增加、凋亡率降低,且呈时间依赖性(P <0.05)。与D组相比,E组成纤维细胞增殖率显著较低,凋亡率较高(P <0.05);(2)与A组相比,B组和C组成纤维细胞中羟脯氨酸含量以及Ⅰ型胶原、Ⅲ型胶原、Wnt2、Wnt3a、β-catenin表达水平均显著降低,而D组均显著增加(P<0.05)。E组羟脯氨酸含量和Ⅰ型胶原、Ⅲ型胶原、Wnt2、Wnt3a、β-catenin水平均显著低于D组(P <0.05);(3)结果表明,脂肪间充质干细胞可以抑制硬皮病小鼠皮损组织成纤维细胞的增殖和胶原沉积,且这一过程与其对Wnt/β-catenin信号通路的抑制作用有关。Adipose-derived mesenchymal stem cells(ADSCs)maintain the potential of multidirectional differentiation during long-term culture in vitro,and can secrete various anti-fibrotic cytokines or growth factors that are involved in regulating fibroblast proliferation and inhibiting collagen deposition,and thereby exert anti-fibrotic effects.OBJECTIVE:To investigate the effects of ADSCs on the proliferation and apoptosis of fibroblasts in scleroderma mice and to explore the underlying mechanisms.METHODS:Fibroblasts were isolated and purified from the skin tissue of scleroderma mice.After passage,the cells were divided into five groups according to different treatment methods:control group,ADSCs intervention group,Wnt inhibitor HY-15597 treatment group(Wnt inhibitor group),Wnt activator SB-216763 treatment group(Wnt activator group),ADSCs+SB-216763 treatment group(combined group).Cell proliferation rate and apoptosis rate were detected using MTT and flow cytometry.The levels of collagen type I,collagen type III,and Wnt signaling pathway related proteins were detected and compared.RESULTS AND CONCLUSION:Compared with the control group,the proliferation rate of fibroblasts in the ADSCs intervention group and Wnt inhibitor group decreased and the apoptotic rate of fibroblasts increased significantly,while the proliferation rate increased and the apoptosis rate decreased in the Wnt activator group in a time-dependent manner(P<0.05).Compared with the Wnt activator group,the proliferation rate of fibroblasts in the combined group was significantly lowered and the apoptotic rate was significantly increased(P<0.05).Compared with the control group,the expression of hydroxyproline,collagen type I,collagen type III,Wnt2,Wnt3a,andβ-catenin in the fibroblasts of ADSCs intervention group and Wnt inhibitor group were significantly decreased,while those in the Wnt activator group were significantly increased(P<0.05).The levels of hydroxyproline,collagen type I,collagen type III,Wnt2,Wnt3a,andβ-catenin in the combined group were signifi

关 键 词：硬皮病 脂肪间充质干细胞 成纤维细胞 细胞增殖 Wnt/β-catenin信号通路 干细胞 皮肤和结缔组织疾病 成纤维细胞 脂肪组织 间质干细胞 WNT蛋白质类 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]