目的:探讨 XMU-MP-1(Xiamen University-inhibitor of mammalian sterile20-like kinase protein 1)对氧糖剥夺(OGD)损伤后小胶质细胞 M1/M2极化平衡的调节作用.方法 采用OGD法诱导BV2细胞损伤.实验分为6组:对照组、模型组、MST1/2 siRNA组和低、中、高剂量实验组(分别给予1.25、5.0和20.0 µg·mL-1 XMU-MP-1).采用MTT法测细胞活力,ELISA测细胞上清液中TNF-α、IL-6和IL-1β表达,qRT-PCR测M1和M2标志物的mRNA表达,流式细胞术测CD206表达,蛋白印迹法测MST1、LATS1和YAP蛋白表达.结果 与模型组相比,XMU-MP-1抑制BV2细胞增殖,显著降低TNF-α、IL-6和IL-1β的表达水平,下调MCP-1、IL-6、TNF-α和iNOS mRNA的表达,上调CD206、IL-10、TGF-β、IL-10和YM1 mRNA表达,降低MST1和LAST1蛋白表达,上调YAP和CD206表达.结论 XMU-MP-1通过调控MST1/2的磷酸化,调节OGD损伤后的BV2细胞M1/M2极化平衡,为神经炎症靶点药物研发提供理论基础.
XMU-MP-1 Protects BV2 Microglia Cells after Oxygen-glucose Deprivation Injury by Regulating the Balance of M1/M2 Polarization
OBJECTIVE To investigate the regulatory effect of XMU-MP-1 on M1/M2 balance in microglia af-ter oxygen glucose deprivation(OGD)injury.METHODS OGD method was used to induce BV2 cell damage.The experiment was divided into 6 groups:control group,model group,MST1/2 siRNA group,and low,medium,and high-dose group(1.25,5.0,and 20.0 µg·mL-1 XMU-MP-1).Using MTT method to measure cell viability and ELISA to measure the expression of TNF-α,IL-6 and IL-1 β.qRT-PCR was used to measure the mRNA expression of M1 and M2 markers.Flow cytometry was used to measure the expression of CD206,and Western blotting was used to measure the expression of MST1,LATS1,and YAP proteins.RESULTS Compared with the model group,XMU-MP-1 inhibi-ted BV2 cell proliferation and significantly reduced the levels of TNF-α,IL-6 and IL-1 β,downregulated the levels of MCP-1,IL-6,TNF-α and iNOS mRNA,upregulated the levels of CD206,IL-10,TGF-β,IL-10 and YM1 mRNA,de-creased the expression of MST1 and LAST1 protein,and upregulated the expression of YAP and CD206 converse-ly.CONCLUSION XMU-MP-1 regulates the phosphorylation of MST1/2 and regulates the M1/M2 balance of BV2 cells after OGD injury,providing a theoretical basis for the development of targeted drugs for neuroinflammation.
万方数据
维普
