脂肪间充质干细胞治疗系统性硬化症模型小鼠的机制  

Therapeutic mechanism of adipose mesenchymal stem cells in mice with systemic sclerosis

作　　者：张晶莹 孙晓林 耿立霞[1] Zhang Jingying;Sun Xiaolin;Geng Lixia(Department of Intensive Medicine,First Affiliated Hospital of Baotou Medical College,Inner Mongolia University of Science&Technology,Baotou 014010,Inner Mongolia Autonomous Region,China;Key Autoimmunity Laboratory of Inner Mongolia,Institute of Rheumatology,Department of Rheumatology,First Affiliated Hospital of Baotou Medical College,Baotou 014010,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古科技大学包头医学院第一附属医院重症医学科,内蒙古自治区包头市014010 [2]包头医学院第一附属医院风湿免疫科(包头医学院风湿免疫研究所),内蒙古自治区自体免疫学重点实验室,内蒙古自治区包头市014010

出　　处：《中国组织工程研究》2021年第13期2011-2017,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81860294),项目负责人:孙晓林;内蒙古自然科学基金项目(2019MS08055),项目负责人:孙晓林;内蒙古科技计划项目(201802089),项目负责人:耿立霞。

摘　　要：背景:虽然已有临床研究发现,自体脂肪间充质干细胞可以有效减轻辐射诱导的系统性硬化症患者面部的纤维化,但其作用机制尚未被深入分析。目的:探讨脂肪间充质干细胞对博来霉素诱导的系统性硬化症模型小鼠的作用机制。方法:取SPF级6-8周龄C57BL/6J雌鼠40只随机分为正常对照组、脂肪间充质干细胞治疗组、博莱霉素组和PBS组,后3组每隔1 d皮下注射博莱霉素,持续28 d,建立系统性硬化症小鼠模型。造模成功后脂肪间充质干细胞组给予皮下注射脂肪间充质干细胞治疗,PBS组皮下注射PBS,均持续治疗14 d。通过ELISA法检测各组小鼠血清中白细胞介素17、转化生长因子β、白细胞介素6和肿瘤坏死因子α水平,苏木精-伊红染色和Masson染色观察系统性硬化症小鼠皮肤和肺组织病变,免疫荧光法检测小鼠皮肤和肺组织中胶原蛋白Ⅰ、Ⅲ、Ⅴ及CD31的表达水平。结果与结论:①与博莱霉素组相比,脂肪间充质干细胞治疗组白细胞介素17、转化生长因子β、白细胞介素6和肿瘤坏死因子α的表达水平显著降低(P<0.01);②与正常对照组相比,博莱霉素组小鼠皮肤真皮层增厚,炎症细胞浸润,皮肤附属器减少;肺泡萎缩塌陷,有许多炎性细胞浸润,肺小动脉壁增厚、微血管基底膜增厚及类纤维蛋白坏死,经脂肪间充质干细胞治疗后炎性症状有所改善;③与正常对照组相比,博莱霉素组小鼠的皮肤和肺组织中胶原纤维大量聚集,经脂肪间充质干细胞治疗后胶原纤维减少;④经脂肪间充质干细胞治疗后,博莱霉素组小鼠皮肤和肺组织中胶原蛋白Ⅰ、Ⅲ、Ⅴ表达水平降低,皮肤组织中CD31表达增多(P<0.01);⑤结果表明,脂肪间充质干细胞可调节博莱霉素小鼠的免疫反应,具有抗纤维化、缓解炎症和减轻血管病变的作用。BACKGROUND: Although clinical studies have found that autologous adipose mesenchymal stem cells can effectively reduce facial fibrosis in patients with radiation-induced systemic sclerosis, but the mechanism of action has not been thoroughly analyzed. OBJECTIVE: To study the mechanism of action of adipose mesenchymal stem cells on bleomycin-induced systemic sclerosis in mice. METHODS: Forty SPF C57BL/6J female mice aged 6-8 weeks were randomly assigned to normal control group, adipose mesenchymal stem cells group, bleomycin group, and PBS group. Mice in the latter three groups were subjected to subcutaneous injection with bleomycin every other day for 28 days, and mouse models of systemic sclerosis were established. After successful model establishment, mice in the adipose mesenchymal stem cells group were subcutaneously injected with adipose mesenchymal stem cells;mice in the PBS group were subcutaneously injected with PBS;the treatments lasted for 14 days. Enzyme-linked immunosorbent assay was used to determine the levels of serum interleukin-17, transforming growth factor-β, interleukin-6, and tumor necrosis factor-α. Hematoxylin-eosin staining and Masson staining were utilized to measure histopathological changes in the skin and lung of systemic sclerosis mice. Immunofluorescence method was applied to examine collagen Ⅰ,Ⅲ and Ⅴ and CD31 expression levels in the skin and lung. RESULTS AND CONCLUSION:(1) Compared with the bleomycin group, the expression levels of interleukin-17, transforming growth factor-β, interleukin-6, and tumor necrosis factor-α were significantly decreased in the adipose mesenchymal stem cells group(P < 0.01).(2) Compared with the normal control group, the skin dermis of mice was thickened;inflammatory cells infiltrated;skin appendages reduced;the alveoli were atrophic and collapsed;with a lot of inflammatory cell infiltration, pulmonary arteriole wall thickening, microvascular basement membrane thickening, and fibrinoid necrosis, and the inflammatory symptoms improved after tre

关 键 词：干细胞 脂肪间充质干细胞 抗炎 抗纤维化 免疫反应 系统性硬化症 小鼠 实验 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]