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骨髓间充质干细胞条件培养基对INS-1细胞凋亡的影响
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骨髓间充质干细胞条件培养基对INS-1细胞凋亡的影响
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Journal Article
骨髓间充质干细胞条件培养基对INS-1细胞凋亡的影响
2015
Overview
目的探讨骨髓间充质干细胞条件培养基（BMSC-CM）对促炎因子诱导大鼠胰岛素瘤细胞（INS-1）凋亡的影响及其作用机制。方法培养大鼠骨髓间充质干细胞（BMSCs）至第3代,收集、浓缩其上清液,获得富含BMSCs分泌因子的条件培养基。用含促炎因子TNF-α、IL-1β、IFN-γ的培养基处理INS-1细胞48h,造成炎症损伤β细胞模型,加入不同浓度的BMSC-CM继续培养12h,所获细胞分为以下5组：正常对照组,促炎因子组,1×BMSC-CM组,2×BMSC-CM组,4×BMSC-CM组。采用CCK-8法检测INS-1细胞存活率,膜连蛋白V/碘化丙啶（Annexin V/PI）检测细胞凋亡率,二氯荧光素双醋酸盐（DCFH-DA）检测细胞内活性氧（ROS）的生成量。结果与正常对照组相比,促炎因子组INS-1细胞存活率下降至正常对照组的75.84%±1.53%,细胞凋亡率增加（17.23%±1.77%）,细胞内ROS水平明显升高（34.3%±0.80%）,差异均具有统计学意义（P〈0.01）。而BMSC-CM有效对抗了促炎因子导致的INS-1细胞损伤,其作用呈剂量依赖性,4×BMSC-CM组效果最佳,该组INS-1细胞存活率较促炎因子组升高了11.86%,达到87.7%±2.08%,细胞凋亡率下降至8.67%±1.59%,ROS生成量减少至17.1%±2.14%,差异均具有统计学意义（P〈0.05）。结论 BMSC-CM可抑制促炎因子诱导的INS-1细胞凋亡,其作用机制可能与BMSC-CM降低细胞内ROS水平有关。
