温肾方对高ALT水平慢性乙型肝炎肾阳虚证患者TLR4/IL-6信号通路的影响

Effect of Wenshen Prescription on TLR4/IL-6 Signaling Pathway in Chronic Hepatitis B Patients with High ALT Level and Kidney Yang Deficiency Syndrome

目的：研究中药温肾方对高丙氨酸氨基转移酶（alanine aminotransferase,ALT）水平慢性乙型肝炎肾阳虚证患者Toll样受体4（toll-like receptor 4,TLR4）/白细胞介素-6（interleukin-6,IL-6）信号通路的影响。方法：选取成都中医药大学附属医院2013年7月至2014年5月收治的68例高ALT慢性乙型肝炎患者,采用前瞻性、平行对照方法设计,将入组病例按1∶1比例随机分组,各34例。治疗组给予温肾方＋替比夫定;对照组予安慰剂＋替比夫定。疗程均为24周。分别在0,12,24周检测外周血乙型肝炎病毒（HBV）DNA,HBV标志物,IL-6;采用实时荧光定量PCR检测外周血单个核细胞TLR4 mRNA的表达;流式细胞学技术检测辅助性T细胞17（T helper cells 17,Th17）/调节性T细胞（regulatory T cells,Treg）。结果：两组基线资料具有可比性。随着治疗时间延长,两组外周血HBV DNA阴转率,HBe Ag阴转率及血清转换率均依次递增。治疗结束后,治疗组在外周血HBV DNA阴转率,HBe Ag阴转率及血清转换率方面均优于对照组（P〈0.05）。治疗过程中,治疗组外周血IL-6水平,TLR4 mRNA水平及Th17/Treg均呈先升高后降低趋势,在12周达到顶点,明显高于本组治疗前（P〈0.05）。治疗12,24周后,治疗组外周血IL-6,TLR4 mRNA水平及Th17/Treg均高于对照组（P〈0.05）。结论：温肾方可提高肾阳虚型高ALT水平慢性乙型肝炎患者的抗病毒疗效,其机制可能是提高TLR4活性,促进IL-6/STAT3信号转导通路表达而调控Th17/Treg平衡。

Objective： To observe the effect of Wenshen prescription on toll-like receptor 4(TLR4）/interleukin-6(IL-6） signaling pathway in chronic hepatitis B(HBV） patients with high ALT level and kidney Yang deficiency syndrome. Method： A total of 68 CHB patients with high ALT level treated in our hospital from July 2013 to May 2014 were selected as research objectives,with 34 cases in each group. Eligible patients were given Wenshen prescription combined with telbivudine(treatment group） or placebo combined with telbivudine(control group） for 24 weeks. Peripheral blood hepatitis B virus DNA,HBV markers,IL-6 were measured at week0,12 and 24. The mRNA expression of TLR4 in peripheral blood mononuclear cell(PBMC） was measured by Real-time PCR,and helper T cells(Th17）/regulatory T cells(Treg） ratio was measured by flow cytometry.Result： There was no significant difference at baseline between the two groups. With the increase of treatment time,HBV DNA negative conversion rate,HBe Ag negative conversion rate and seroconversion rate of the two groups were gradually decreased. At week 24,treatment group was superior to control group in the rates of HBV DNA negative conversion,HBe Ag negative conversion and seroconversion. During the treatment,the peripheral blood IL-6,TLR4 mRNA levels and Th17/Treg ratio first increased and later decreased in the treatment group.They reached the peak at week 12,which were significantly higher than the baseline(P〈0. 05）. At week 12 and24,the peripheral blood IL-6,TLR4 mRNA and Th17/Treg in treatment group were superior to those in control group(P〈0. 05）. Conclusion： Wenshen prescription has an increased antiviral effect on telbivudine-treated CHB patients with high ALT level and kidney Yang deficiency syndrome,and its mechanism may be related to the regulation of Th17/Treg balance by TLR4/IL-6/STAT3 signaling pathway.