炎症激活骨髓间充质干细胞条件培养基对辐射损伤IEC-6细胞的修复  

Bone marrow mesenchymal stem cell conditioned medium with inflammatory activation for repair of radiation-induced IEC-6 cell injury

作　　者：李金亮 骆昱均 马发鑫 郑跃[2] 卢铨 周伟杰 张宸 沙卫红 陈浩[2] Li Jinliang;Luo Yujun;Ma Faxin;Zheng Yue;Lu Quan;Zhou Weijie;Zhang Chen;Sha Weihong;Chen Hao(Second Clinical Medical College of Southern Medical University, Guangzhou 510000, Guangdong Province, China;Department of Digestion, Guangdong General Hospital (Guangdong Academy of MedicalSciences), Guangzhou 510000, Guangdong Province, China)

机构地区：[1]南方医科大学第二临床医学院,广东省广州市510000 [2]广东省人民医院消化内科(广东省医学科学院),广东省广州市510000

出　　处：《中国组织工程研究》2019年第5期697-702,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81741067);项目负责人:陈浩;广东省自然科学基金(2016A030313815);项目负责人:沙卫红;广州市科技计划项目(201707010419);项目负责人:沙卫红;广州市科技计划项目(201804010050);项目负责人:陈浩~~

摘　　要：背景:间充质干细胞条件培养基是干细胞移植理想的替代方案,课题组前期研究表明炎症预激活的间充质干细胞条件培养基能促进辐射损伤小肠组织学结构和功能的修复,改善急性辐射损伤大鼠生存状态,但其潜在的细胞机制未进一步探讨。目的:观察炎症预激活的间充质干细胞条件培养基对辐射损伤小肠上皮细胞增殖、凋亡的影响,探讨预激活间充质干细胞条件培养基修复小肠黏膜的细胞机制。方法:小肠隐窝IEC-6细胞分为对照组、辐射损伤组、辐射损伤+MSC-CM^(IEC-6(NOR))组和辐射损伤+MSC-CM^(IEC-6(IR))组,后3组以10 Gy X射线一次性照射IEC-6细胞建立体外急性小肠辐射损伤模型,对照组及辐射损伤组加入DMEM-F12培养基,辐射损伤+MSC-CM^(IEC-6(NOR))组加入正常状态间充质干细胞条件培养基,辐射损伤+MSC-CM^(IEC-6(IR))组加入炎症激活状态间充质干细胞条件培养基。照射后3 d收集细胞行免疫荧光PCNA染色观察增殖改变,行TUNEL凋亡染色及Western blot蛋白免疫印迹法观察凋亡改变。结果与结论:与辐射损伤组相比,辐射损伤+MSC-CM^(IEC-6(IR))组IEC-6细胞PCNA阳性细胞数明显增加(P<0.05),TUNEL阳性细胞数下降(P<0.05),凋亡相关因子Caspases-3蛋白表达明显下降(P<0.05),而辐射损伤+MSC-CM^(IEC-6(NOR))组的各项指标变化无统计学意义(P>0.05)。上述结果表明,与正常状态的间充质干细胞条件培养基相比,炎症激活的间充质干细胞条件培养基显著促进辐射损伤肠上皮细胞的增殖,并抑制其凋亡,促进辐射损伤小肠上皮修复。BACKGROUND:Conditioned medium from mesenchymal stem cells(MSC-CM)may represent a promising alternative to MSCs transplantation.Our previous study has demonstrated that MSC-CM with inflammatory activation improves the structural and functional restoration of the small intestine after radiation-induced intestinal injury,improve the survival status of rats with acute radiation injury,but its potential cellular mechanism has not been further explored.OBJECTIVE:To observe the effect of MSC-CM with inflammatory activation on the proliferation and apoptosis of intestinal epithelial cells(IEC-6)after radiation injury and to investigate the cellular mechanism of pre-activated MSC-CM in repairing the small intestinal mucosa.METHODS:IEC-6cells were divided into four groups:control group,radiation injury group,normal MSC-CM(MSC-CM^IEC-6(NOR))group and inflammatory pre-activated MSC-CM(MSC-CM^IEC-6(IR))group.IEC-6cells in the latter three groups were exposed to10Gy X-ray irradiation and cultured in DMEM-F12medium,MSC-CMIEC-6(IR)and MSC-CM^IEC-6(NOR)respectively.Cells in the control group were only cultured in DMEM-12medium.Cultured cells were collected at3days after radiation to observe the proliferation of IEC-6cells by using proliferating cell nuclear antigen immunofluorescence staining,and to observe the apoptosis by using TUNEL apoptosis staining and western blot assay.RESULTS AND CONCLUSION:Compared with the radiation injury group,in the MSC-CM^IEC-6(IR)group,the number of cells positive for proliferating cell nuclear antigen increased significantly(P<0.05),the number of TUNEL positive cells decreased significantly(P<0.05),and the expression of Caspases-3decreased significantly(P<0.05),However,there was no significant difference between the MSC-CM^IEC-6(NOR)group and radiation injury group(P>0.05).Taken together,MSC-CM^IEC-6(IR),but not non-activated MSC-CM,significantly promotes the proliferation and reduces apoptosis of intestinal epithelial cells after radiation injury and therefore repair the injured intestinal tissu

关 键 词：间充质干细胞条件培养基 骨髓间充质干细胞 炎症 条件培养基 小肠上皮细胞 放射性肠损伤 增殖 凋亡 修复 国家自然科学 间质干细胞 培养基 条件性 小肠 辐射损伤 组织工程 

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