前列腺素E2促进乙型肝炎病毒复制的机制研究

摘要/Abstract

摘要： 目的探讨前列腺素E2(PGE2)对乙型肝炎病毒(HBV)复制及T细胞功能的影响。方法 ELISA法检测慢性乙肝患者(CHB,120例)和健康体检者(HP,60例)血清中PGE2的浓度。检测PGE2高水平或低水平患者血清中丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)的水平,流式细胞术检测血样中CD8⁺T细胞功能相关指标。将24只经尾静脉注射重组病毒rAAV8-1.3HBV的小鼠随机均分为3组,每组8只:对照组(DMSO)、PGE2组(PGE2的稳定类似物)和PGE2受体拮抗组(PGE2受体拮抗剂),试剂盒及实时定量PCR检测3组小鼠血清中HBsAg、HBV DNA及pgRNA水平,流式细胞术检测肝脏中CD8⁺T细胞功能相关指标。结果与HP(201.3±13.38)对比,CHB患者(877.2±43.27)血清中PGE2水平显著增加(P<0.05)。与PGE2低水平患者相比,PGE2高水平患者血清中HBV DNA(1.84±0.161)、ALT(722.64±33.081)、AST(514.20±13.427)水平显著增加,CD8⁺T细胞中Tim3的表达显著增加(18.64±2.051),颗粒酶B的表达显著降低(0.29±0.102)(P<0.05)。与对照组相比,PGE2组小鼠血清中HBsAg(5.57±0.941)、HBV DNA(0.58±0.073)及pgRNA(0.57±0.069)水平显著增加,肝脏CD8⁺ T细胞中Tim3(25.45±3.412)的表达显著增加,TNF-α(21.50±3.139)和IFNγ(15.23±2.105)的表达显著降低(P<0.05)。PGE2受体拮抗有相反的效果。结论 PGE2在CHB患者血清中呈高表达,PGE2促进HBV复制及T细胞衰竭,PGE2信号阻断不仅可修复T细胞功能还可控制HBV复制。

关键词: 前列腺素E2, 乙型肝炎病毒, 复制, 机制

Abstract: Objective To investigate the effect of prostaglandin E2 (PGE2) on hepatitis B virus (HBV) replication and T cell function.Methods The concentrations of PGE2 in serum of chronic hepatitis B (CHB) patients and healthy persons (HP) were measured by enzyme-linked immunosorbent assay. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum of patients with high or low levels of PGE2 were detected by kit,according to the instructions from the manufacturers. Flow cytometry was used to detect the function of cluster of differentiation 8⁺T cell (CD8⁺T cell) in blood samples. A total of 24 mice injected with recombinant adeno-associated virus 8 carrying 1.3 copies of HBV genome via tail vein were randomly divided into 3 groups: control group (dimethylsulfoxide), PGE2 group (stable analog of PGE2) and PGE2 receptor antagonist group (PGE2 receptor antagonist). The levels of hepatitis B surface antigen (HBsAg), HBV deoxyribonucleic acid (DNA) and pregenomic ribonucleic acid (pgRNA) in mouse serum of the 3 groups were detected by kit and real-time quantitative polymerase chain reaction. The function of CD8⁺ T cell in the liver were detected by flow cytometry. Results The serum level of PGE2 in CHB patients was significantly higher than that in HP (877.2 ± 43.27 vs. 201.3 ± 13.38, P<0.05). In patients with high PGE2, the serum levels of HBV DNA, ALT, AST were significantly higher than those in patients with low PGE2 (1.84 ± 0.161 vs. 0.92 ± 0.143, 722.64 ± 33.081 vs. 246.95 ± 26.629, 514.20 ± 13.427 vs. 183.07 ± 12.259, P<0.05), and the expression of T cell immunoglobulin and mucin domain 3 in blood CD8⁺ T cells (Tim3) was significantly higher (18.64 ± 2.051 vs. 15.12 ± 2.265, P<0.05), the expression of granzyme B was significantly lower than that in patients with low PGE2 (0.29 ± 0.102 vs. 0.38 ± 0.095, P<0.05). Compared with the control group, the serum levels of HBsAg, HBV DNA and pgRNA in the PGE2 group were significantly higher (5.57 ± 0.941 vs. 4.51 ± 0.662, 0.58 ± 0.073 vs. 0.42 ± 0.049, 0.57 ± 0.069 vs. 0.40 ± 0.051, P<0.05), the expression of Tim3 in liver CD8⁺ T cells was significantly higher (25.45 ± 3.412 vs. 12.63 ± 2.313, P<0.05), and the expression of tumor necrosis factor-α and interferon-γ was significantly lower in the PGE2 group (21.50 ± 3.139 vs. 27.36 ± 3.841, 15.23 ± 2.105 vs. 24.81 ± 2.595, P<0.05). PGE2 receptor antagonism has the opposite effect.Conclusion PGE2 is highly expressed in serum of CHB patients, which promotes replication of HBV and depletion of T cell. And blocking of PGE2 can not only restore T cell function but also reduce HBV replication.

Key words: Prostaglandin E2, Hepatitis B Virus, Replication, Mechanism

黄珊珊, 赵静雅, 陈洁, 杨扬. 前列腺素E2促进乙型肝炎病毒复制的机制研究[J]. 肝脏, 2020, 25(3): 277-281.

HUANG Shan-shan, ZHAO Jing-ya, CHEN Jie, YANG Yang.. The study on mechanism of prostaglandin E2 promoting replication of hepatitis B virus[J]. Chinese Hepatolgy, 2020, 25(3): 277-281.

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