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、MyD88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、MyD88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、MyD88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-MyD88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 pre-treated 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 ex-pression of TLR4 /MyD88 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, MyD88 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/MyD88 protein in dose-dependent manner. Conclusion DFMG may play an important role in protecting endothelial cells from inflammatory in-jury 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-MyD88 signal transduction.
展开▼
