重型肝炎患者外周血Toll样受体信号通路负性调节因子基因表达的初步研究

Preliminary study on negative regulator gene expression of toll-like receptor signal pathways in patients with acute-on-chronic liver failure associated with hepatitis B virus infection

摘要:

目的：探讨Toll样受体（toll-like receptors，TLRs）信号转导通路负性调控因子在重型乙型肝炎患者免疫发病机制中的作用。方法：采用荧光定量PCR检测慢性乙型肝炎组20例、重型肝炎早期组17例，重型肝炎中晚期组9例患者及健康对照组18例外周血单个核细胞TLRs信号通路负性调节因子髓样分化蛋白88短臂（myeloid-differentiation-88-short，MyD88s）、白介素1受体相关激酶-M（interleukin-1R-associated-kinase-M，IRAK-M）、单免疫球蛋白白细胞介素1受体相关蛋白（single-im－munoglobulin-interleukin-1R-related-molecule，SIGIRR）、锌指蛋白A20及TLR4基因的表达；ELISA法检测血浆肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白介素-10（interleukin-10，IL-10）的水平。结果：与健康对照组相比，MyD88s、IRAK-M、A20的基因表达水平及血清TNF-α、IL-10水平随病情加重逐渐升高（P<0.05）；而IL-10在慢性乙型肝炎组与重型肝炎早期组比较，差异无统计学意义（P >0.05）；TLR4 mRNA在慢性乙型肝炎组、重型肝炎早期组、重型肝炎中晚期组均高于健康对照组，重型肝炎中晚期组低于早期组（P<0.05）。相关性分析显示，慢性乙型肝炎组、重型肝炎早期组TLR4 mRNA与同组TNF-α、总胆红素水平正相关（P<0.05），与凝血酶原活动度呈负相关（P<0.05）。结论：TLRs信号通路负性调节因子参与慢性肝炎和重型肝炎的发生，且负性调节因子持续升高可能导致重型肝炎患者免疫抑制。

Abstract:

Objective：To explore the role of negative regulators of toll-like receptor（TLRs） signal pathways in immunological patho-genesis of severe hepatitis B virus infection（CHB）. Methods：mRNA expressions of myeloid-differentiation-88-short（MyD88s），interleukin-1R-associated-kinase-M（IRAK-M），single-immunoglobulin-interleukin-1R-related-molecule（SIGIRR），zinc finger protein A20 and TLR4 in peripheral blood mononuclear cells（PBMCs） in 20 cases of chronic CHB，17 cases of acute-on-chronic liver failure at early stage（ACLF-E），9 cases of acute-on-chronic liver failure at late stage（ACLF-L） and 18 healthy controls were detected by real time fluorescence quantitative PCR. Serum levels of tumor necrosis factor-α（TNF-α） and interleukin-10（IL-10） were detected by ELISA assay. Results：Compared with those in healthy controls，mRNA expression levels of MyD88s，IRAK-M and A-20 as well as serum levels of TNF-α and IL-10 were unregulated with the progression of diseases in CHB patients，ACLF-E patients and ACLF-L patients（P<0.05）（P<0.01）. There was no significant difference in serum levels of IL-10 between CHB and ACLF-E（P >0.05）. mRNA expressions of TLR4 were higher in CHB patients，ACLF-E patients and ACLF-L patients than in healthy controls. mRNA expressions of TLR4 were lower in ACLF-L patients than in ACLF-E patients（P<0.05）. Conclusions：Negative regulators of TLRs signaling pathway participate in the pathogenesis of chronic hepatitis and liver failure and upregulation of negative regulators may lead to immunosuppressive in patients with liver failure.