DFMG调节TLR4-MyD88信号转导保护内皮细胞受损

Effect of DFMG on TLR4-MyD88 signal transduction in vascular endothelial cells injured by LPC

目的:探索7-二氟甲氧基-5,4’-二甲氧基金雀异黄(DFMG)对溶血磷脂胆碱(LPC)诱导的人脐静脉内皮细胞(HUVE-12)炎症损伤的影响及可能的作用机制。方法:用不同浓度的LPC处理HUVE-12细胞,选择最适合浓度的LPC制备炎症损伤模型,DFMG、TLR4特异性拮抗剂(CLI-095)预处理HUVE-12细胞后加最适浓度的LPC,通过MTT法检测HUVE-12的增殖,流式细胞术检测细胞的凋亡,ELISA检测各组细胞培养液中TNF-α的表达、乳酸脱氢酶(LDH)的活性和western blot检测各组HUVE-12细胞TLR4、My D88蛋白的表达水平。结果:LPC抑制HUVE-12增殖,促进HUVE-12细胞LDH的释放和TNF-α分泌,增加细胞的凋亡,上调其TLR4、My D88蛋白表达,DFMG呈浓度依赖性的拮抗LPC对HUVE-12增殖的作用,抑制细胞LDH的释放和TNF-α的分泌,拮抗TLR4、My D88蛋白表达上调。结论:DFMG具有拮抗LPC引发的内皮细胞炎性损伤作用,作用机制可能是抑制TLR4-My D88信号转导。

Objective To explore the effect of DFMG on the inflammatory injury of human umbilical vein endothelial cells(HUVE-12) induced by LPC and its possible mechanism. Methods HUVE-12 was deal with different concentrations of LPC and the appropriate concentration of LPC was chosen to prepare cell inflammatory injury model. HUVE-12 cells were pretreated with DFMG or a specific antagonist of TLR4(CLI-095) then deal with optimal concentration of LPC, the proliferation of HUVE-12 was detected by MTT, the expression level of TNF-α and LDH in the culture was detected by ELISA and the expression of TLR4/My D88 protein were measured western blot. Results LPC inhibited the proliferation of HUVE-12 cells and promoted the secretion of TNF-α, the release of LDH and expression of TLR4, My D88 protein. DFMG inhibited the effect of LPC on the proliferation of HUVE-12, the secretion of TNF-α, the release of LDH and expression of TLR4/My D88 protein in dose-dependent manner. Conclusion DFMG may play an important role in protecting endothelial cells from inflammatory injury induced by LPC. The mechanism of DFMG on protecting endothelial cells from oxidative injury in HUVEC-12 cells maybe probably associate with inhibiting the TLR4-My D88 signal transduction.