不同时相HBV-ACLF患者外周血Treg/Th17细胞变化及与HBV-DNA相关性研究  

Changes in peripheral blood Treg/Th17 cells and their correlation with HBV-DNA in HBV-ACLF patients at different phase

作　　者：杜珊 谭年花 彭杰[1] 曾孟晖 陈斌[1] DU Shan;TAN Nianhua;PENG Jie;ZENG Menghui;CHEN Bin(First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan University of Chinese Medicine)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]湖南中医药大学

出　　处：《中国病原生物学杂志》2024年第10期1117-1122,共6页Journal of Pathogen Biology

基　　金：国家中医药管理局高水平中医药重点学科建设项目(zyyzdxk-2023146);国家自然科学基金面上项目(No.81673959);湖南省高层次卫生人才“225”工程学科带头人培养项目(湘卫函[2019]196号);湖南省中医药科研计划项目重点课题(No.C2022005);湖南中医药大学校院联合基金项目(No.2022XYLH003)。

摘　　要：目的 研究不同时相乙肝相关性慢加急性肝衰竭(HBV-ACLF)患者外周血Treg/Th17细胞表达差异,以及与HBV-DNA水平的相关性。方法 60例HBV-ACLF患者分为不同时相,即进展期(Adv-ACLF)28例和平台期(Pla-ACLF)32例,同时纳入慢性乙型肝炎(CHB)患者、健康志愿者(NC)各20例作为对照。流式细胞仪检测各组受试者外周血Treg、Th17细胞频率并计算Treg/Th17比值,流式细胞微球芯片捕获技术(CBA)检测细胞因子转化生长因子(TGF-β)、白介素-10(IL-10)、肿瘤坏死因子(TNF-α)、白介素-17A(IL-17A)、白介素-23(IL-23)水平,实时荧光定量RT-PCR法检测Treg、Th17细胞特异性转录因子FoxP3、ROR-γt mRNA表达;进一步分析不同时相患者Treg、Th17细胞频率及Treg/Th17细胞比值与HBV-DNA水平的相关性。结果 与NC组、CHB组比较,HBV-ACLF进展期及平台期患者炎症指标TNF-α、IL-17A、IL-23水平以及ROR-γtmRNA表达明显升高(P<0.01),Th17细胞频率升高、Treg/Th17值下降(P<0.01),Treg/Th17细胞失衡向Th17方向偏移,与平台期比较进展期更严重(P<0.05);进展期HBV-ACLF患者外周血Treg、Th17细胞频率与HBV-DNA水平呈正相关(Treg r=0.516,P=0.005;Th17 r=0.420,P=0.026),但Treg/Th17比值与HBV-DNA水平无相关性。平台期HBV-ACLF患者外周血Treg、Th17细胞频率以及Treg/Th17比值均与HBV-DNA水平无相关性。结论 HBV-ACLF进展期、平台期患者均存在Treg/Th17细胞失衡,进展期患者Treg/Th17失衡较平台期更严重。进展期HBV-ACLF患者Treg、Th17细胞频率与HBV-DNA水平呈正相关。Objective e This article explores the differential expression of Treg/Th17 cells in hepatitis B associated chronic and acute liver failure(HBV-ACLF)patients with different phase,and the correlation with the level of HBV-DNA.Methods 60 HBV-ACLF patients were divided into different phases,namely 28 patients with advanced-ACLF(Adv-ACLF)and 32 patients with plateau-ACLF(Pla-ACLF),while 20 patients with chronic hepatitis B(CHB)and 20 healthy volunteers(normal control,NC)were used as controls.Flow cytometry was used to detect the frequency of Treg and Th17 cells in the peripheral blood of each group of subjects and calculate the Treg/Th17 ratio.Cytometric Bead Array(CBA)was used to detect the levels of cytokines transforming growth factor-β(TGF-β),interleukin-10(IL-10),tumor necrosisfactor-α(TNF-α),interleukin-17A(IL-17A),and interleukin-23(IL-23).Real-time fluorescence quantitative PCR(real-time PCR)method to detect Treg,Th17 cell specific transcription factor Forkhead box protein P3(FoxP3),Retinoic acid receptor-related orphan receptor gamma t(ROR-yt)mRNA expression.The correlation of Treg,Th17 cell frequency and Treg/Th17 cell ratio with HBV-DNA level was further analyzed.Results Compared with CHB and NC group,the levels of inflammatory markers TNF-α,IL-17A,IL-23,and ROR-t mRNA expression were significantly increased in patients with advanced and plateau stages of HBV-ACLF(P<0.O1),the frequency of Th17 cells increased and the ratio of Treg/Th17 decreased(P<0.01),the imbalance of Treg/Th17 cells shifted towards the Th17 direction,and the advanced stage group was more severe than the plateau stage group(P<0.05).Correlation analysis found that the peripheral blood Treg and Th17 cell frequencies of patients with advanced HBV-ACLF were positively correlated with HBV-DNA level(Treg r=0.516,P=0.005;Th17 r=0.420,P=0.026),but Treg/Th17 ratio has no correlation with HBV-DNA level.The peripheral blood Treg and Th17 cell frequency and Treg/Th17 ratio of patients with HBV-ACLF plateau stage had no correlation with HBV-DNA l

关 键 词：肝衰竭 TREG细胞 TH17细胞 炎症反应 

分 类 号：R512.62[医药卫生—内科学] R575.3[医药卫生—临床医学]