Objective To study the effect of macrophage phenotype and mesenchymal stem cells (MSC) on polarization of macrophage phenotype damaged due to combined high glucose and inflammation.Methods Primarily cultured peritoneal macrophages from rats were divided into normal group,damaged group (33 mmol/L glucose and 1 μg/ml lipopolysaccharide),treatment group (cocultured high glucose + Inflammation-damaged LPS + MSC transwell),and inhibitor group (MSC+COX-2 inhibitor).Morphology of macrophages was observed under optical microscope,M1 and M2 macrophages were detected by flow cytometry,and serum levels of IL-6,IL-10 and PGE2 were measured by ELISA.Results The ratio of M1 macrophages was significantly lower while that of M2 macrophages was significantly higher in treatment group than in damaged group (8.91％±0.71％ vs 16.19％±0.78％,P＜0.05;51.09％±4.89％ vs 28.10％±12.47％,P＜0.05).The serum level of IL-6 was significantly lower while that of IL-10 was significantly higher in treatment grup than in damaged group (P＜0.05).The ratio of M1 macrophages was significantly higher in inhibitor group than in treatment group (19.94％ ± 2.07％ vs 8.91％ ± 0.71％,P＜0.05).The serum level of IL-6 was significantly higher while that of IL-10 was significantly lower in inhibitor group than in treatment group (P＜0.05).Conclusion MSC can promote the M1 macrophaes damaged due to combined high glucose and inflammation to the polarization of M2 macrophage phenotype and repair the damaged macrophages through the COX-2 pathway.%目的 探讨高糖联合炎症条件下巨噬细胞的袁型变化及脂肪间充质干细胞(MSC)对损伤的巨噬细胞表型极化的影响及其机制.方法 将原代培养的大鼠腹腔巨噬细胞分为正常组、损伤组(33 mmol/L葡萄糖+1μg/ml脂多糖)、治疗组(高糖+脂多糖炎症损伤+ MSC Transwell共培养)、抑制剂组[MSC+环氧化酶2(COX-2)抑制剂].光学显微镜观察巨噬细胞的形态,流式细胞术定量检测巨噬细胞M1及M2表型比例,ELISA细胞因子白细胞介素(IL)-6、IL-10、前列腺素E2的变化.结果 与损伤组比较,治疗组M1型巨噬细胞比例明显降低[(8.91±0.71)％ vs (16.19±0.78)％,P＜0.05],M2型巨噬细胞比例明显升高[(51.09±4.89)％ vs (28.10±12.47)％,P＜0.05],IL-6明显下降,IL-10明显升高;与治疗组比较,抑制剂组M1型巨噬细胞的比例显著升高[(19.94±2.07)％ vs (8.91±0.71)％,P＜0.05],IL-6含量增加,IL-10含量降低,差异有统计学意义(P＜0.05).结论 MSC可c通过COX-2通路促进高糖炎症损伤的M1型巨噬细胞向M2表型极化及修复其损伤.
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