槲皮素对PGN诱导的大鼠肠黏膜微血管内皮细胞炎性损伤的作用及其机制研究

The Effect and Mechanism of Quercetin on Peptidoglycan-induced Inflammatory Injury of Rat Intestinal Mucosal Microvascular Endothelial Cells

应用肽聚糖(PGN)诱导大鼠肠黏膜微血管内皮细胞(RIMMVECs)炎性损伤模型,探究槲皮素对RIMMVECs炎性损伤的保护作用并探讨其作用机制。通过CCK8法筛选槲皮素作用浓度,荧光定量PCR(FQ-PCR)法测量Toll样受体2(TLR2)、Toll样受体4(TLR4)的mRNA表达水平,Western Blot法测量TLR2、TLR4和其下游MyD88蛋白表达水平,和ELISA法评估炎性因子TNF-α分泌水平,综合评价槲皮素对PGN诱导的RIMMVECs细胞炎性损伤的保护作用。结果表明,不同浓度的槲皮素能显著降低TLR2和TLR4 mRNA和蛋白水平,降低MyD88蛋白表达水平,进一步降低炎性因子TNF-α释放量(P<0.05)。槲皮素能保护RIMMVECs免受PGN导致的细胞炎性损伤。

In the present study,cellular inflammatory injury model of rat intestinal mucosal microvascular endothelial cells(RIMMVECs)was induced by peptidoglycan(PGN)to investigate the protective potential of quercetin on RIMMVECs inflammatory injury.First,we screened appropriate action concentrations of quercetin using CCK8 assay.We comprehensively evaluated the potential effects of quercetin on RIMMVECs inflammatory injury through the detection of toll-like receptor 2(TLR2)and toll-like receptor 4(TLR4)mRNA expressions using fluorescence quantitative PCR(FQ-PCR)assay.TLR2 and TLR4 and its downstream MyD88 protein expressions were analyzed by Western Blot,and the secretion of inflammatory factor TNF-αwas analyzed by ELISA assay.The results showed that quercetin significantly reduced both the mRNA and protein expression of TLR2 and TLR4,decreased the protein expression of MyD88,and reduced the release of inflammatory factor TNF-α(P<0.05).These data indicate that quercetin can protect RIMMVECs from PGN-induced cellular inflammatory injury.