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

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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（sTRAIL）。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PDC-E2+PDTC组。空白对照组不予任何刺激；PDC-E2组予2、10和50 μg/mL的PDC-E2刺激；PDC-E2+HTA125组予10 μg/mL的TLR4功能抑制抗体HTA125孵育4 h后，再加入50 μg/mL的PDC-E2；PDC-E2+PDTC组加入100 nmol/mL的NF-κB抑制剂PDTC 0.5 h后，再加入50 μg/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sTRAIL浓度。结果：2、10和50 μg/mL浓度PDC-E2刺激U937细胞24 h，TLR4表达均明显增加，但无浓度依赖性。浓度为50 μg/mL的PDC-E2刺激U937细胞1 h，NF-κB即显著活化；至2 h，活化逐渐减少；至4 h，已明显减少。加入HTA125后，NF-κB活性较阻断前显著降低。浓度为50 μg/mL的PDC-E2刺激U937细胞24 h，培养上清TNF-α和sTRAIL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48 h和72 h，TNF-α、IL-12和sTRAIL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sTRAIL。

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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

Abstract：Objective: To explore whether pyruvate dehydrogenase complex E2 subunit (PDC-E2) may activate Toll-like receptor 4 (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-E2 was used to stimulate monocyte line U937, combined with HTA125 (an inhibitory antibody of TLR4) and PDTC (an inhibitor of NF-κB). The study groups included control group, PDC-E2 group, PDC-E2+HTA125 group and PDC-E2+PDTC group. TLR4 on U937 was detected by flow cytometry, activity of NF-κB was determined by EMSA, TNF-α, IL-12 and sTRAIL were measured by ELISA. Results: At 24 h after stimulation by 2, 10 and 50 μg/mL of PDC-E2, TLR4 expression on U937 cell was significantly increased. At 1 h after stimulation by 50 μg/mL of PDC-E2, NF-κB was markedly activated, while at 2 h, the activation of NF-κB was gradually decreased and at 4 h, the decrease was very significant. NF-κB activity was significantly decreased after HTA125 addition compared with that before. At 24 h after stimulation by 50 μg/mL of PDC-E2, supernant TNF-α and sTRAIL levels were significantly higher than that of control, while IL-12 level was of no significant difference. At 24 h and 48 h, the levels of TNF-α, IL-12 and sTRAIL were all significantly higher than that of control. The three cytokines were significantly lower after addition of HTA125 or PDTC than before. Conclusion: PDC-E2 can activate TLR4-NF-κB pathway, by which PDC-E2 stimulates the secretion of TNF-α, IL-12 and sTRAIL by monocyte.

Key words： pyruvate dehydrogenase complex Toll-like receptor 4 nuclear factor cytokine primary biliary cholangitis

收稿日期: 2016-11-23

基金资助:

国家自然科学基金面上项目（81671594，81571591）

作者简介: 杨再兴（1978-），男，辽宁锦州人，研究员，博士

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2017/V47/I9/637

[1] HIRSCHFIELD G M, GERSHWIN M E. The immunobiology and pathophysiology of primary biliary cirrhosis[J]. Annu Rev Pathol, 2013, 8: 303-330.
[2] SASAKI M, KAKUDA Y, MIYAKOSHI M, et al. Infiltration of inflammatory cells expressing mitochondrial proteins around bile ducts and in biliary epithelial layer may be involved in the pathogenesis in primary biliary cirrhosis[J]. J Clin Pathol, 2014, 67(6): 470-476.
[3] LEICESTER K L, OLYNYK J K, BRUNT E M, et al. Differential findings for CD14-positive hepatic monocytes/macrophages in primary biliary cirrhosis, chronic hepatitis C and nonalcoholic steatohepatitis[J]. Liver Int, 2006, 26(5): 559-565.
[4] TSUNEYAMA K, HARADA K, KONO N, et al. Scavenger cells with gram-positive bacterial lipoteichoic acid infiltrate around the damaged interlobular bile ducts of primary biliary cirrhosis[J]. J Hepatol, 2001, 35(2): 156-163.
[5] WANG A P, MIGITA K, ITO M, et al. Hepatic expression of toll-like receptor 4 in primary biliary cirrhosis[J]. J Autoimmun, 2005, 25(1): 85-91.
[6] MAO T K, LIAN Z X, SELMI C, et al. Altered monocyte responses to defined TLR ligands in patients with primary biliary cirrhosis[J]. Hepatology, 2005, 42(4): 802-808.
[7] ZHAO J, ZHAO S, ZHOU G, et al. Altered biliary epithelial cell and monocyte responses to lipopolysaccharide as a TLR ligand in patients with primary biliary cirrhosis[J]. Scand J Gastroenterol, 2011, 46(4): 485-494.
[8] ZHANG Z, SMITH C, LI W, et al. Characterization of nitric oxide modulatory activities of alkaline-extracted and enzymatic-modified arabinoxylans from corn bran in cultured human monocytes[J]. Agric Food Chem, 2016, 64(43): 8128-8137.
[9] DING J, GUO C, HU P, et al. CSF1 is involved in breast cancer progression through inducing monocyte differentiation and homing[J]. Int J Oncol, 2016, 49(5): 2064-2074.
[10] TSENG H H, VONG C T, KWAN Y W, et al. TRPM2 regulates TXNIP-mediated NLRP3 inflammasome activation via interaction with p47 phox under high glucose in human monocytic cells[J]. Sci Rep, 2016, 6: 35016.
[11] WANG R, STEPHENS J, LACY M J. Characterization of monoclonal antibody HTA125 with specificity for human TLR4[J]. Hybrid Hybridomics, 2003, 22(6): 357-365.
[12] BOYD J H, MATHUR S, WANG Y, et al. Toll-like receptor stimulation in cardiomyoctes decreases contractility and initiates an NF-kappaB dependent inflammatory response[J]. Cardiovasc Res, 2006, 72(3): 384-393.
[13] 余国庆, 陈晓明. 多糖免疫受体研究进展[J]. 温州医学院学报, 2012, 42(4): 396-401.
[14] FLOREANI A, INFANTOLINO D, BIASIN R, et al. Tumour necrosis factor alpha and cellular proliferation in primary biliary cirrhosis[J]. Ital J Gastroenterol Hepatol, 1999, 31(1): 56-60.
[15] 闫惠平, 庄辉, 刘燕敏, 等. 原发性胆汁性肝硬化患者的免疫学特点分析[J]. 中华肝脏病杂志, 2015, 13(1): 12-16.
[16] YANG C Y, MA X, TSUNEYAMA K, et al. IL-12/Th1 and IL-23/Th17 biliary microenvironment in primary biliary cirrhosis: implications for therapy[J]. Hepatology, 2014, 59(5): 1944-1953.
[17] LIANG Y, YANG Z, LI C, et al. Characterisation of TNF-related apoptosis-inducing ligand in peripheral blood in patients with primary biliary cirrhosis[J]. Clin Exp Med, 2008,8(1): 1-7.
[18] HARADA K, NAKANUMA Y. Biliary innate immunity: function and modulation[J]. Mediators Inflamm, 2010, 2010(1): 60-68.
[19] ARNDTZ K, HIRSCHFIELD G M. The pathogenesis of autoimmune liver disease[J]. Dig Dis, 2016, 34(4): 327-333.