TLR4/RhoA信号通路调控连续血液滤过治疗引起脓毒症内皮细胞通透性变化的机制  

Mechanism of TLR4/RhoA signaling pathway in endothelial cell permeability changes induced by continuous hemofiltration therapy in sepsis

作　　者：余慧林 王建丰 刘义[1] 刘玉尧 蒋薇 孟承颖 王欢[1] 胡德林[1] Yu Huilin;Wang Jianfeng;Liu Yi;Liu Yuyao;Jiang Wei;Meng Chengying;Wang Huan;Hu Delin(Dept of Burn and Wound Repair,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]安徽医科大学第一附属医院烧伤与创面修复外科,合肥230022

出　　处：《安徽医科大学学报》2023年第7期1159-1164,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省重点研究和开发计划项目(编号:1804h08020230);安徽省省级临床重点专科建设项目(编号:060102027)。

摘　　要：目的探讨Toll样受体4(TLR4)/Ras同源基因家族成员A(RhoA)信号通路参与调控连续血液滤过前后脓毒症血清对血管内皮细胞通透性影响的分子机制。方法收集5例经连续血液滤过治疗前后的脓毒症患者血清,检测滤过前后血清中炎症细胞因子含量;应用滤过前后血清干预人脐静脉内皮细胞(HUVEC)24 h,检测经连续滤过治疗前后的血清干预对于血管内皮细胞钙黏蛋白(VE-cadherin)、聚合肌动蛋白丝(F-actin)、TLR4和RhoA表达的影响;构建TLR4低表达细胞株,检测TLR4低表达对血管内皮细胞钙黏蛋白(VE-cadherin)、F-actin和RhoA表达的影响及其对内皮细胞通透性的影响。结果滤过后血清中TLR4、RhoA、白细胞介素-1(IL-1)、白细胞介素-6(IL-6)和肿瘤坏死因子α(TNF-α)含量均显著降低;滤过后血清干预组VE-cadherin、F-actin、TLR4和RhoA的表达水平均显著降低,同时细胞通透性明显降低;TLR4低表达可明显促进VE-cadherin和F-actin的表达,抑制RhoA蛋白的表达。结论TLR4/RhoA信号通路参与了连续血液滤过治疗引起脓毒症血清对血管内皮细胞通透性变化的调节。Objective To investigate the molecular mechanism of Toll-like receptor 4(TLR4)/Ras homologue A(RhoA)signaling pathway involved in regulating the effect of septic serum on vascular endothelial cell permeability before and after continuous hemofiltration.Methods The serum of 5 patients with sepsis before and after continuous hemofiltration treatment was collected,and the levels of inflammatory cytokines in serum before and after hemofiltration were detected.Human umbilical vein endothelial cells(HUVEC)were treated with serum before and after continuous hemofiltration for 24 hours.The expression of VE-cadherin,F-actin,TLR4 and RhoA in vascular endothelial cells were detected by Western blot.A TLR4 low expression cell line was constructed to detect the effect of TLR4 low expression on the expression of VE-cadherin,F-actin and RhoA and the permeability of endothelial cells.Results After continuous blood treatment,the serum levels of TLR4,RhoA,interleukin-1(IL-1),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)significantly decreased.The expression levels of VE-cadherin,F-actin,TLR4 and RhoA in the serum intervention group after continuous hemofiltration treatment significantly decreased,and the cell permeability significantly decreased.Low expression of TLR4 significantly promoted the expression of VE-cadherin and F-actin,and inhibited the expression of RhoA protein.Conclusion TLR4/RhoA signaling pathway is involved in the regulation of changes in vascular endothelial cell permeability induced by septic serum after continuous hemofiltration treatment.

关 键 词：脓毒症 TLR4 RHOA 连续血液滤过治疗 内皮细胞 烧伤 

分 类 号：R644[医药卫生—外科学]