丙酮酸脱氢酶复合体E2亚基通过活化Toll样受体4影响单核细胞分泌细胞因子  

Biological effect of pyruvate dehydrogenase complex E2 subunit on monocyte U937 by activating Toll-like receptor 4-NF-κB pathway

作　　者：杨再兴[1] 梁艳[2] 刘东红[1] 张治宇 仲人前[2] YANG Zaixing;LIANG Yan;LIU Donghong;ZHANG Zhiyu;ZHONG Renqian(Department of Laboratory Medicine, Taizhou First People’s Hospital, Taizhou, 318020;Department of Laboratory Diagnostics, Shanghai Changzheng Hospital, Shanghai, 200003;Department of VIP, Shanghai Changzheng Hospital, Shanghai, 200003)

机构地区：[1]台州市第一人民医院检验科,浙江台州318020 [2]上海长征医院实验诊断科,上海200003 [3]上海长征医院贵宾科,上海200003

出　　处：《温州医科大学学报》2017年第9期637-641,共5页Journal of Wenzhou Medical University

基　　金：国家自然科学基金面上项目(81671594;81571591)

摘　　要：目的:探讨丙酮酸脱氢酶复合体E2亚基(PDC-E2)是否可以活化Toll样受体4(TLR4)-NF-κB通路,并通过该通路诱导单核细胞分泌肿瘤坏死因子α(TNF-α)、白介素12(IL-12)和可溶性肿瘤坏死因子相关凋亡诱导配体(s TRAIL)。方法:培养单核细胞株U937,分为空白对照组、PDC-E2组、PDC-E2+HTA125组和PDCE2+PDTC组。空白对照组不予任何刺激;PDC-E2组予2、10和50μg/mL的PDC-E2刺激;PDC-E2+HTA125组予10μg/mL的TLR4功能抑制抗体HTA125孵育4h后,再加入50μg/mL的PDC-E2;PDC-E2+PDTC组加入100nmol/mL的NF-κB抑制剂PDTC0.5h后,再加入50μg/mL的PDC-E2。通过流式细胞术检测TLR4表达,EMSA实验检测NF-κB活化情况,ELISA法检测培养上清TNF-α、IL-12和s TRAIL浓度。结果:2、10和50μg/mL浓度PDC-E2刺激U937细胞24h,TLR4表达均明显增加,但无浓度依赖性。浓度为50μg/mL的PDC-E2刺激U937细胞1h,NF-κB即显著活化;至2h,活化逐渐减少;至4h,已明显减少。加入HTA125后,NF-κB活性较阻断前显著降低。浓度为50μg/mL的PDC-E2刺激U937细胞24h,培养上清TNF-α和s TRAIL的浓度显著高于空白对照组(P<0.05),IL-12与空白对照组比差异无统计学意义;至48h和72h,TNF-α、IL-12和s TRAIL的浓度均显著高于空白对照组(P<0.05)。加入HTA125或PDTC后,各时间点3种细胞因子的浓度均较阻断前显著降低(P<0.05)。结论:PDC-E2可以活化TLR4-NF-κB通路,并通过该通路刺激单核细胞分泌细胞因子TNF-α、IL-12和s TRAIL。Objective:To explore whether pyruvate dehydrogenase complex E2subunit(PDC-E2)mayactivate Toll-like receptor4(TLR4)-NF-κB pathway and induce secretion of tumor necrosis factor-α(TNF-α),interleukin-12(IL-12)and TNF-related apoptosis-inducing ligand(sTRAIL).Methods:PDC-E2was used tostimulate monocyte line U937,combined with HTA125(an inhibitory antibody of TLR4)and PDTC(an inhibitorof NF-κB).The study groups included control group,PDC-E2group,PDC-E2+HTA125group and PDCE2+PDTC group.TLR4on U937was detected by flow cytometry,activity of NF-κB was determined by EMSA,TNF-α,IL-12and sTRAIL were measured by ELISA.Results:At24h after stimulation by2,10and50μg/mLof PDC-E2,TLR4expression on U937cell was significantly increased.At1h after stimulation by50μg/mL ofPDC-E2,NF-κB was markedly activated,while at2h,the activation of NF-κB was gradually decreased and at4h,the decrease was very significant.NF-κB activity was significantly decreased after HTA125addition comparedwith that before.At24h after stimulation by50μg/mL of PDC-E2,supernant TNF-αand sTRAIL levels weresignificantly higher than that of control,while IL-12level was of no significant difference.At24h and48h,thelevels of TNF-α,IL-12and sTRAIL were all significantly higher than that of control.The three cytokines weresignificantly lower after addition of HTA125or PDTC than before.Conclusion:PDC-E2can activate TLR4-NF-κB pathway,by which PDC-E2stimulates the secretion of TNF-α,IL-12and sTRAIL by monocyte

关 键 词：丙酮酸脱氢酶复合体 TOLL样受体4 核因子 细胞因子 原发性胆汁性胆管炎 

分 类 号：R392.32[医药卫生—免疫学]