缺氧诱导骨髓间充质干细胞条件培养基体外修复辐射损伤小肠上皮细胞变化  

Bone marrow mesenchymal stem cell conditioned medium with hypoxic activation enhances its effects on radiation-induced intestinal epithelial cell injury in vitro

作　　者：郑跃[1] 陈浩[1] 沙卫红[1] 王启仪[1] 刘婉薇[1] 

机构地区：[1]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510080

出　　处：《中国组织工程研究》2015年第45期7230-7236,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);广东省科技计划项目(2015A020212029)~~

摘　　要：背景:间充质干细胞条件培养基含有丰富的间充质干细胞旁分泌物质,被认为是干细胞移植理想的替代方案。然而,正常状态的间充质干细胞条件培养基可能由于旁分泌活性不足,常无法有效修复损伤组织。目的:观察缺氧诱导的骨髓间充质干细胞条件培养基(MSC-CM^(Hyp))对大鼠小肠隐窝上皮IEC-6细胞辐射损伤后增殖和凋亡的影响,并进一步探讨其修复小肠上皮细胞的旁分泌机制。方法:IEC-6细胞接受10 Gy X射线照射后分别加入MSC-CM^(Hyp)、正常培养的骨髓间充质干细胞条件培养基(MSC-CM^(Nor)组)及DMEM-F12培养基(DMEM-F12组)培养。结果与结论:锥虫蓝染色、流式细胞仪、Western blot检测结果显示,与DMEM-F12组相比,MSC-CM^(Hyp)组IEC-6细胞存活数和增殖率明显增加(P<0.05),凋亡率和凋亡相关因子Caspases-3/8表达明显下降(P<0.05),而MSC-CM^(Nor)组的各项指标与DMEM-F12组比较差异无显著性意义(P>0.05)。EILSA检测结果显示,与MSC-CM^(Nor)相比,MSC-CM^(Hyp)中血管内皮生长因子、碱性成纤维细胞生长因子、胰岛素样生长因子1、和白细胞介素10的含量明显升高(P<0.05)。结果表明,缺氧诱导的骨髓间充质干细胞条件培养基中细胞因子含量明显升高,显著促进辐射损伤IEC-6细胞的增殖,同时下调细胞凋亡信号,从而进一步促进损伤细胞的修复。BACKGROUND： Conditioned medium from mesenchymal stem cells（MSC-CM） that contains abundant MSCs paracrine substances may represent a promising alternative to MSCs transplantation. However, normal MSC-CM with insufficient paracrine ability is not effective for tissue damage repair.OBJECTIVE： To investigate the effects of MSC-CM with（MSC-CM-（Hyp）） and without hypoxic activation（MSC-CM-（Nor）） on the proliferation and apoptosis of radiation-induced injured intestinal epithelial cells（IEC-6） and to further discuss the paracrine mechanisms. METHODS： IEC-6 cells were exposed to 10 Gy irradiation and cultured in MSC-CM-（Hyp）, MSC-CM-（Nor）, and DMEM-F12 medium, respectively. RESULTS AND CONCLUSION： Findings from trypan blue staining, flow cytometry and western blot assay showed that, compared with the DMEM-F12 medium group, treatment with MSC-CM-（Hyp） significantly enhanced IEC-6 viability proliferation after radiation-induced injury, as well as significantly decreased cell apoptosis and expression of Caspases-3/8（P〈0.05）. However, there was no significant difference between the MSC-CM-（Nor） group and DMEM-F12 medium group（P〉0.05）. On the other hand, the increased levels of vascular endothelial growth factor, basic fibroblast growth factor, insulin-like growth factor-1, and interleukin-10 were detected in the MSC-CM-（Hyp） group compared to the MSC-CM-（Nor） group（P〈0.05）. These results suggest that the MSC-CM-（Hyp） improves the viability and proliferative capacity of IEC-6 cells after radiation-induced injury via up-regulating secretion of cytokines and down-regulating apoptotic signaling.

关 键 词：间充质干细胞 培养基 条件性 旁分泌 小肠 组织工程 干细胞 骨髓干细胞 条件培养基 骨髓间充质干细胞 小肠损伤 细胞治疗 移植 缺氧诱导 小肠上皮细胞 细胞因子 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]