摘要
目的探讨脂肪间充质干细胞(MSCs)培养上清对高糖、炎症环境中巨噬细胞表型的影响及其机制。方法分别以及联合利用高浓度葡萄糖(33 mmol/L)、脂多糖(LPS;1μg/ml)体外处理大鼠腹腔巨噬细胞12 h。依据是否采用MSCs干预分为干预组和非干预组,考察各损伤条件下两组巨噬细胞表型的差异。MSCs干预方法为加入MSCs上清0.5 ml,培养48 h。结果在各非干预组中,与正常条件下相比,各损伤条件下的M1型巨噬细胞比例、M1/M2值和IL-6水平均显著增高,而M2型巨噬细胞比例以及IL-10和PGE2水平均显著降低。MSCs干预后,在LPS和高糖+LPS的损伤条件下,与非干预组相比,干预组的M1型巨噬细胞的比例和M1/M2值均显著降低,而M2型巨噬细胞的比例均增加,ELISA结果也显示,在各损伤条件下,与非干预组相比,干预组的IL-6水平均显著降低,而IL-10和PGE2的水平均显著增加。结论高糖及慢性炎症因素叠加可加剧巨噬细胞向M1促炎表型极化及炎症因子的分泌,MSCs上清作用于损伤的巨噬细胞,促使其向M2抗炎表型极化,同时使炎症因子分泌减少,抗炎因子分泌增多。
Objective To determine the effect of the supernatant of adipose-derived mesenchymal stem cells （AD-MSCs） on macrophages phenotype cultured under high glucose or combined with inflammation circumstance.Methods Rat peritoneal macrophages were isolated and then cultured under 33 mmol/L glucose,1 μg/ml LPS and their combination separately for 12 h, then the culture media were added with 0.5 ml supernatant of AD-MSCs.After 48h,the morphology of the macrophages with the supernatant intervention or not was observed with microscopy.The percentages of M1 and M2 macrophages were detected by flow cytometry, and the concentrations of cytokines IL-6, IL-10 and PGE2 were determined with ELISA.Results Compared with normal control cells, the percentage of M1 macrophage,M1/M2 ratio and IL-6 concentration were significantly higher, while the proportion of M2 macrophages and contents of IL-10 and PGE2 were obviously declined in the macrophages treated with high glucose, LPS or their combination.However, addition of the MSCs supernatant resulted in notable decrease in the percentage of M1 macrophages and M1/M2 ratio, and significant increases in the proportion of M2 macrophage in the LPS and combination groups.The results of ELISA indicated that MSCs supernatant reduced the level of IL-6 while elevated those of IL-10 and PGE2 in the macrophages treated with high glucose, LPS or their combination.Conclusion Combination of high glucose and LPS synergistically aggravates the polarization of pro-inflammatory M1 macrophages, and promotes the secretion of inflammatory cytokines.The supernatant of MSCs improves M2 phenotype polarization and production of anti-inflammatory cytokines.
作者
靳丽媛
邓子辉
张金英
杨晨
司艺玲
陈光辉
JIN Li-Yuan DENG Zi-Hui ZHANG Jin-Ying YANG Chen SI Yi-Ling CHEN Guang-Hui(Department of Cardiology, 2Institute of Basic Medicine, Chinese PLA General Hospital, Beijing 100853, China)
出处
《中华老年多器官疾病杂志》
2017年第4期283-287,共5页
Chinese Journal of Multiple Organ Diseases in the Elderly
基金
国家高技术研究发展计划(863计划
2011AA020101)
国家自然科学基金面上项目(81471052)~~
关键词
间充质干细胞
巨噬细胞极化
高糖
慢性炎症
mesenchymal stem cells
macrophage polarization
high glucose
chronic inflammation
