摘要
目的 探讨利多卡因对脂多糖(lipopolysaccharide,LPS)诱导的脓毒症肾损伤的保护作用及机制.方法 将肾小管上皮细胞(HK-2)分为空白对照组、LPS组和LPS+利多卡因组.细胞增殖与毒性试剂(cell counting kit-8,CCK-8)法检测细胞增殖活性,采用酶联免疫吸附试验(ELISA)检测各组HK-2细胞炎症因子高迁移率族蛋白B1(high mobility group protein B1,HMGB1)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达水平,比较各组间上述指标的差异,采用蛋白质免疫印迹试验(Western blotting)检测各组HK-2细胞关键蛋白Toll样受体4(Toll-like receptor 4,TLR4)、核因子κB p65(NF-KB p65)的表达.结果 CCK-8检测结果显示,20 μg/mL的利多卡因能使LPS诱导24 h后的HK-2细胞生存率提高73％,LPS诱导HK-2细胞24 h后,TLR4、NF-κB p65蛋白表达均显著高于空白对照组,HMGB1、IL-6、TNF-a水平也均显著高于空白对照组;而与LPS组比较,20 μg/mL利多卡因可明显降低TLR4、NF-κB p65的蛋白表达,也能明显降低HMGB1、IL-6、TNF-a水平.结论 20 μg/mL利多卡因可显著降低LPS诱导的HK-2细胞中促炎性细胞因子的释放,同时还可显著下调炎症通路蛋白TLR4、NF-κB的表达,从而发挥对LPS诱导的SA-AKI模型HK-2细胞的保护作用,其作用机制可能与抑制TLR4/NF-κB信号通路有关.
Abstract
Objective To investigate the protective effect and mechanism of lidocaine on lipopolysaccharide(LPS)-induced sepsis-associated acute kidney injury.Methods The renal tubular epithelial cells(HK-2)were divided into blank control group,LPS group and LPS+lidocaine group.Cell proliferation and toxicity reagents(cell counting kit-8,CCK-8),The expression levels of HK-2 cell inflammatory factor(high mobility group protein B1,HMGB 1),interleu-kin-6(interleukin-6,IL-6),and tumor necrosis factor-α(tumor necrosis factor-α,TNF-α)were measured by enzyme-linked immunosorbent assay(ELISA),Comparing the differences in the above indicators between the groups,Western blotting was used to detect the key protein Toll-like receptor 4(Toll-like receptor 4,TLR4),the expression of the nu-clear factor κB p65(NF-κB p65).Results The CCK-8 test showed,Lidocaine with 20 μg/mL improved the survival of HK-2 cells after 24 h of LPS induction by 73％,In HK-2 cells for 24 h after LPS induction,TLR4 and NF-κB p65 pro-tein expression were significantly higher than the blank control group,The levels of HMGB1,IL-6,and TNF-a were al-so significantly higher than those of the blank control group;While,when compared with the LPS group,20 μg/mL li-docaine could significantly reduce the protein expression of TLR 4,NF-κB p65,It also significantly reduced the levels of HMGB1,IL-6,and TNF-a.Conclusion 20 μg/mL lidocaine can significantly reduce the release of proinflammatory cy-tokines in HK-2 cells,and also significantly downregulate the expression of inflammatory pathway proteins TLR4 and NF-κB,thus exerting the protection effect of LPS-induced HK-2 cells,and the mechanism of action may be related to the inhibition of TLR4/NF-κB signaling.
基金项目
新疆维吾尔自治区卫生健康青年医学科技人才专项科研项目(WJWY-202325)
维普
万方数据