乙型肝炎肝硬化患者辅助性T淋巴细胞17和调节性T淋巴细胞及维甲酸相关核孤儿受体γt和叉状头/翅膀状螺旋转录因子3 mRNA表达水平的变化及其临床意义研究

Changes of Th17 and Treg and the Expressions of RORγt and Foxp3 mRNA in Patients With HBV Related Liver Cirrhosis and the Clinical Significance

目的通过对乙型肝炎肝硬化患者外周血辅助性T淋巴细胞17(Th17)、调节性T淋巴细胞(Treg)及其特异性转录因子维甲酸相关核孤儿受体γt(RORγt)和叉状头/翅膀状螺旋转录因子3(Foxp3)mRNA表达水平的检测探讨其临床意义。方法选取2015年7—12月昆明医科大学第一附属医院诊治的慢性乙型肝炎病毒(HBV)感染者64例,其中慢性乙型肝炎(CHB)患者21例(CHB组)、代偿期肝硬化(CLC)患者21例(CLC组)、失代偿期肝硬化(DCLC)患者22例(DCLC组)。同期选取本院体检健康者20例为对照组。收集研究对象一般资料〔性别、年龄及丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆红素(TBIL)〕;应用流式细胞术检测研究对象外周血Th17、Treg表达率,计算Th17/Treg比值;应用实时荧光定量反转录聚合酶链式反应(RT-q PCR)法测定外周血RORγt、Foxp3 mRNA表达水平。结果 4组年龄、ALT、AST、TBIL比较,差异有统计学意义(P<0.05)。CHB组、CLC组、DCLC组Th17、Treg表达率及Th17/Treg比值均高于对照组(P<0.05);DCLC组Th17、Treg表达率及Th17/Treg比值均高于CHB组,Th17表达率及Th17/Treg比值均高于CLC组(P<0.05)。CHB组、CLC组、DCLC组RORγt mRNA及Foxp3 mRNA表达水平均高于对照组(P<0.05);DCLC组RORγt mRNA及Foxp3 mRNA表达水平均高于CHB组、CLC组(P<0.05)。CHB组、CLC组、DCLC组外周血Th17表达率与RORγt mRNA表达水平均呈正相关(P<0.05);CHB组、CLC组、DCLC组外周血Treg表达率与Foxp3 mRNA表达水平均呈正相关(P<0.05);CHB组、CLC组、DCLC组外周血Th17表达率、RORγt mRNA表达水平与ALT呈正相关(P<0.05);CHB组外周血Foxp3mRNA表达水平与ALT呈正相关(P<0.05);CHB组RORγt mRNA表达水平、Foxp3 mRNA表达水平与AST、TBIL呈正相关(P<0.05)。结论 Th17、Treg及其特异性转录因子RORγt、Foxp3可能与乙型肝炎肝硬化患者的疾病进展与肝功能有关。

Objective To analyze the changes of peripheral Th17 and Treg and the expressions of RORγt and Foxp3 mRNA in patients with HBV related liver cirrhosis and explore the clinical significance. Methods We enrolled 64 patients with chronic HBV in fection who received treatment in the First Affiliated Hospital of Kunming Medical University from July to December in 2015. Among them,21 patients with chronic hepatitis B were assigned into CHB group,21 patients at compensated liver cirrhosis were assigned into CLC group,and 22 patients at decompensated liver cirrhosis were assigned into DCLC group. 20 healthy people who received physical examination in the hospital in the same period were enrolled as control group. The general data（gender,age,ALT,AST and TBIL）were collected;expression rates of Th17 and Treg in the peripheral blood of the subjects were detected using flow cytometry,and the ratio of Th17 / Treg was calculated. RT-qPCR method was applied to determine the expressions of RORγt and Foxp3 mRNA of peripheral blood. Results The four groups were significantly different in age,ALT,AST and TBIL（ P 〈 0. 05）. CHB group,CLC group and DCLC group were higher than control group in the expression rates of Th17 and Treg and Th17 / Treg（P 〈 0. 05）;DCLC group was higher than CHB group in the expression rates of Th17 and Treg and Th17 / Treg,and was higher than CLC group in the expression rate of Th17 and Th17 / Treg（P 〈 0. 05）. CHB group,CLC group and DCLC group were higher than control group in the expressions of RORγt mRNA and Foxp3 mRNA （P 〈 0. 05）;DCLC group was higher than CHB and CLC group in the expressions of RORγt mRNA and Foxp3 mRNA（P 〈0. 05）. For CHB group,CLC group and DCLC group,the expression rate of Th17 was positively correlated with the expression of RORγt mRNA（ P 〈 0. 05）;for CHB group,CLC group and DCLC group,the expression rate of Treg was positively correlated with the expression of Foxp3 mRNA（P 〈 0. 05）;for CHB group,CLC group and DCLC group,the expression rate of Th17 and the expression of RORγt mRNA was positively correlated with ALT（P 〈 0. 05）;for CHB group,the expression of Foxp3 mRNA was positively correlated with ALT（ P 〈 0. 05）;for CHB group,the expressions of RORγt mRNA and Foxp3 mRNA were positively correlated with AST and TBIL（ P 〈 0. 05 ） . Conclusion Th17 and Treg cells and their specific transcription factors RORγt and Foxp3 may be related with the progress of HBV - related liver cirrhosis and liver function.