摘要
目的研究慢性乙型肝炎(CHB)患者外周Th17/调节性T细胞(Treg)失衡中IL-21的作用及其初步机制。方法流式细胞仪分别检测59例CHB患者和20例健康人外周血细胞中Th17细胞和Treg细胞频数,同时以ELISA法检测血清细胞因子IL-21、IL-17、IL-10、TGF-β水平。体外分别以IL21、IL-2刺激CHB患者外周血单核细胞(PBMC),以实时PCR检测培养细胞转录因子RORγt mRNA及Foxp3 mRNA表达水平,以ELISA法检测培养上清细胞因子IL-17、IL-10和TGF-β水平。结果与正常人群相比,IL-21、IL1 7、TGF-β水平显著升高(P均<0.05),CHB患者外周Th17/Treg比例显著升高(P<0.05),IL-21水平与Th1 7/Treg比例呈正相关(P=01.008)。与IL-2相比.IL-21刺激CHB患者PBMC的RORγt mRNA表达水平明显升高(P=0.016)而Foxp3 mRNA表达水平明显降低(P=0.018),相对应的培养上清中,IL-1 7水平显著上升(P=0.049)而IL-10、TGF-β水平明显降低(P=0.017,P=0.004)。结论 C、HB患者中IL-21能通过提高RORγt mRNA表达,同时抑制Foxp3 mRNA表达进而促进Th1 7/Treg失衡。
Objective To investigate the role of interleukin-21 in T helper cell 17(Thl7)/regulatory T cells(Treg)imbalance in chronic hepatitis B(CHB) patients,and its mechanism.Methods Frequencies of Thl7 cells and Treg in peripheral blood mononuclear cells(PBMCs) of 76 CHB patients and 20 health controls were detected by flow cytometry,and serum levels of interleukin(ID-21.IL-17.transforming growth factor(TGF)-β and IL-10 were measured by enzymelinked immune-sorbent assay(ELISA).Under stimulation of IL-21 or IL-2 in vitro,levels of RORyt and Foxp3 mRNA in PBMCs were detected by real time reverse transcription polymerase chain reaction(RT-PCR),and levels of IL-17.IL-10.TGF-β in supernatants were determined by ELISA assay,respectively.Results The level of IL-21 was positively associated with the ratio of Th17 to Treg.which was significantly enhanced in CHB patients when compared with that in health controls(P〈0.05).In IL-21 stimulated PBMCs of CHB patients,the expression of RORyt mRNA was significantly increased(P = 0.016).while the expression of Foxp3 mRNA was obviously decreased(P = 0.018).comparing with those in IL-2 stimulation group.Additionally,level of IL-17 in the corresponding supernatants was notably enhanced(P =0.049),while levels of IL-10 and TGF-p were decreased significantly(P = 0.017.P = 0.004).respectively.Conclusion IL-21 could enhance the ratio of Th17 to Treg through increasing the expression of RORyt mRNA and decreasing the expression of Foxp3 mRNA in patients with CHB.
出处
《肝脏》
2016年第4期259-262,312,共5页
Chinese Hepatology
基金
江苏省临床医学科技专项(BL2012043)