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.
出处
《中国组织工程研究》
CAS
北大核心
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)~~
