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目的观察人脐带来源的间充质干细胞(h UC-MSCs)在正常环境和炎症环境下对小胶质细胞活化的调节作用,探讨h UC-MSCs对中枢神经系统炎症反应的免疫调节作用。方法将BV2细胞与人脐带间充质干细胞来源的条件培养基(h UC-MSCs-CM)和(或)脂多糖(LPS)共培养24或48 h后利用MTT比色法分析BV2增殖活力的变化,ELISA试剂盒分析BV2细胞上清中TNF-α和IL-6的变化,免疫印迹法检测精氨酸酶1(Arg1)的表达,Real-time PCR分析TNF-α、IL-6、ARG1基因的表达水平。结果 1 BV2在LPS长程刺激下表现出明显的增殖效应,而h UC-MSCs-CM可以抑制这种增殖反应;2炎症刺激诱导BV2表达TNF-α和IL-6明显增多,而在h UC-M SCs-CM和LPS共刺激时TNF-α和IL-6表达明显低于LPS单刺激组;3h UC-M SCs-CM共培养组BV2细胞高表达M2型小胶质细胞标志物Arg1。结论脐带间充质干细胞抑制炎症所致的小胶质细胞增殖;脐带间充质干细胞通过旁分泌机制调节小胶质细胞向M2型活化,降低促炎因子表达,这可能是其最终发挥神经保护作用的机制之一。
OBJECTIVE: To observe the regulation of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) on microglial activation in normal environment and inflammatory environment, and to explore the immunomodulatory effect of hUC-MSCs on central nervous system inflammatory response. Methods BV2 cells were co-cultured with human umbilical cord mesenchymal stem cell-derived conditioned medium (hUC-MSCs-CM) and / or lipopolysaccharide (LPS) for 24 or 48 h, and the viability of BV2 The changes of TNF-α and IL-6 in supernatant of BV2 cells were analyzed by ELISA kit. The expression of Arg1 was detected by Western blotting. The expression of TNF-α, IL-6, ARG1 Gene expression levels. Results 1 BV2 showed obvious proliferative effect under long-term stimulation of LPS, h UC-MSCs-CM could inhibit this proliferation reaction.2 Inflammatory stimuli induced BV2 expression of TNF-α and IL-6 significantly increased, while in h UC- The expression of TNF-α and IL-6 in M SCs-CM and LPS co-stimulated group was significantly lower than that in LPS monolayer group. BV2 cells in UC-M SCs-CM co-culture group were highly expressed M2 microglia marker Arg1 in 3h. Conclusion Umbilical cord mesenchymal stem cells inhibit proliferation of microglial cells induced by inflammation. Umbilical cord mesenchymal stem cells regulate the activation of microglia to M2 and reduce the expression of proinflammatory cytokines through paracrine mechanism, which may be the result of neuroprotection One of the mechanisms of action.