炎症预激活骨髓间充质干细胞条件培养基修复小肠黏膜急性辐射损伤  

Effects of bone marrow mesenchymal stem cell conditioned medium with and without inflammatory activation on radiation-induced intestinal injury

作　　者：刘婉薇[1,2] 陈韵[3] 郑跃[1,2] 沙卫红[1,2] 王启仪[1,2] 叶山亮[1,2] 陈浩[1,2] 

机构地区：[1]广东省人民医院消化内科 [2]广东省医学科学院,广东省广州市510080 [3]广东医学院第一临床学院,广东省湛江市524023

出　　处：《中国组织工程研究》2015年第10期1544-1550,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81300279);中国博士后科学基金项目(2012M521579);广东省自然科学基金项目(S2013040013549);广东省医学科研基金项目(A2014020)~~

摘　　要：背景:间充质干细胞条件培养基被认为是干细胞移植的良好替代方案。既往研究表明,炎症因子的诱导激活能增强间充质干细胞多种生物学潜能,而正常状态的间充质干细胞可能由于免疫活性和迁徙能力不足,无法有效修复损伤组织。目的:探讨炎症预激活前后骨髓间充质干细胞条件培养基对急性辐射损伤小肠上皮治疗作用的差异。方法:分离、培养、鉴定SD幼鼠骨髓间充质干细胞,分别与正常对照和辐射损伤的小肠隐窝细胞株IEC-6在Transwell培养板中共培养24 h,预刺激的骨髓间充质干细胞继续单独培养48 h,收集到的上清液分别为正常状态间充质干细胞条件培养基(MSC-CM NOR)和辐射炎症预激活间充质干细胞条件培养基(MSC-CM IR)。将成年SD大鼠随机分为4组,正常对照组、辐射损伤组、MSC-CM NOR治疗组和MSC-CM IR治疗组,以14 Gy剂量一次性腹部局部照射制备急性小肠辐射损伤大鼠模型,模型制备后尾静脉注射和植入式胶囊渗透压泵腹腔植入联合给药。于治疗后第1,3,7天取小肠组织检测短回路电流和血清木糖水平观察小肠分泌、吸收功能变化。治疗后第3天取小肠组织行电镜观察超微结构改变,治疗后第1,3,5,7,14天取小肠组织行苏木精-伊红染色观察小肠黏膜组织学改变。记录各组大鼠生存状态及生存时间。结果与结论:输注MSC-CM IR后,大鼠小肠短回路电流差值、血清木糖水平较辐射损伤组升高,差异有显著性意义(P<0.05)。苏木精-伊红染色和电镜观察显示从治疗后第3天起MSC-CM IR组小肠黏膜病理改变明显减轻,上皮厚度、紧密连接间隙明显优于辐射损伤组。生存分析显示MSC-CM IR组生存率和平均生存时间明显改善(P<0.05),而MSC-CM NOR组的各项指标与辐射损伤组比较差异无显著性意义(P>0.05)。结果表明炎症激活状态下的骨髓间充质干细胞条件培养基可促进辐射损伤小肠结构和功能�BACKGROUND： Conditioned medium from mesenchymal stem cells（MSC-CM） may represent a promising alternative to MSCs transplantation. Previous studies have shown that inflammatory activation can strengthen the multiple biological potencies of MSCs; however, normal MSCs with insufficiency of immunocompetence and migration ability are not effective for tissue damage repair.OBJECTIVE： To investigate differential effects of MSC-CM with and without inflammatory activation on radiation-induced intestinal injury.METHODS： MSCs from the bone marrow of SD rats were separated, cultured and identified, and then co-cultured with non-irradiated IEC-6 or irradiated IEC-6 in a transwell system for 24 hours. Then, MSCs with inflammatory activation were cultured alone for another 48 hours. After that, the supernatant was collected as non-activated MSC-CM（MSC-CM^NOR） and MSC-CM under radiation-induced inflammatory condition（MSC-CM^IR）. Rats were exposed to 14 Gy whole abdominal irradiation and randomly divided into four groups： control group, radiation injury group（DMEM/F12）,MSC-CM^NOR group and MSC-CM^IR group. Continuous administration was given via tail vein and intraperitoneal implantation of Alzet microosmotic pumps. Intestinal samples were collected at 1, 3, 7 days after radiation for analysis of short circuit variation, at 3 days after radiation for analysis of intestinal epithelium ultrastructure, and at 1, 3, 5, 7, 14 days after radiation for histological observation of the intestinal epithelium using hematoxylin-eosin staining. Blood samples were collected at 1, 3, 7 days after radiation for analysis of serum xylose levels. In addition, the survival state and survival time of rats were observed and recorded.RESULTS AND CONCLUSION： The short circuit variation responding to electrical field stimulation was significantly reduced at all frequencies, but it was significantly improved in the MSC-CM^IR group. Similarly, the intestinal absorption（serum xylose levels） was also significantly impaired by ir

关 键 词：骨髓 间质干细胞移植 培养基 条件性 辐射损伤 小肠 干细胞 移植 骨髓间充质干细胞 条件培养基 细胞疗法 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]