慢性乙型肝炎中医证型与HLA-DR13基因、基本核心启动子突变及T细胞亚群变化的相关性研究

Correlation Study on Chinese Medical Syndrome Types of Chronic Hepatitis B Patients and HLA-DR13 Gene,BCP Mutation,and T-lymphocyte Subsets

目的探讨慢性乙型肝炎中医临床各证型与HLA-DR13基因、基本核心启动子(BCP)突变(即A1762T/G1764A)及T细胞亚群变化的相关性。方法 102例乙型肝炎患者按中医辨证分型分为肝胆湿热、肝郁脾虚、肝肾阴虚、瘀血阻络及脾肾阳虚证型5种,设30名健康人为正常对照组。实时荧光定量PCR检测血液中的乙肝病毒核酸定量(HBV-DNA)水平及HLA-DR13基因,流式细胞仪检测T淋巴细胞CD4+、CD8+表达,测序法检测样本血清BCP变异,ELISA方法检测乙肝病毒e抗原,并分析各证型客观指标的相关性。结果 HBV-DNA定量结果各证型之间差异均无统计学意义(P>0.05);HBe Ag阳性率各组间比较,肝郁脾虚型HBe Ag阳性率高于与其他各证型(P<0.05),即肝郁脾虚型>肝肾阴虚型>瘀血阻络型>肝胆湿热型>脾肾阳虚型。与正常对照组比较,脾肾阳虚型CD3+及CD3+CD4+值明显降低(P<0.05),肝胆湿热型及脾肾阳虚型CD3+CD4+/CD3+CD8+值亦降低(P<0.05),各证型CD3+CD8+值差异均无统计学意义(P>0.05)。HLA-DR13基因定量与正常对照组比较,肝郁脾虚型和肝肾阴虚型组降低(P<0.05);BCP突变以肝肾阴虚型阳性率高于其他各证型,差异有统计学意义(P<0.05)。结论 HLA-DR13和BCP检测结果可作为临床慢性乙型肝炎中医辨证分型的参考指标。

Objective To explore the correlation between the HLA-DR13,basic core promoter（ BCP）,changes of T lymphocyte subset and clinical Chinese medical syndromes of chronic hepatitis B（ CHB）. Methods Totally102 CHB patients were syndrome typed as Gan depression Pi deficiency syndrome（ GDPDS）,Pi-Shen yang deficiency syndrome（ PSYDS）, Gan-gallbladder dampness heat syndrome（ GGDHS）, Gan-Shen yin deficiency syndrome（ GSYDS）,and static blood blocking collaterals syndrome（ SBBCS）. Besides,30 healthy subjects were recruited as the normal control group. The blood HBV-DNA level and HLA-DR13 gene were detected with real time fluorescent PCR. The expression of CD4^＋and CD8^＋in T lymphocytes was detected using flow cytometry. The mutation of serum A1762 T / G1764 A was detected using PCR sequencing. Hepatitis Be antigen（ HBe Ag） was detected with ELISA,and correlation between various Chinese medical syndrome types and objective indicators were analyzed. Results There was no statistical difference in HBV-DNA quantitative results among various syndrome types（ P 〉0. 05）. HBe Ag positive rate was higher in GDPDS than in other syndrome types（ P 〈0. 05）. It was sequenced as GDPDS 〉GSYDS〉 SBBCS〉 GGDHS 〉PSYDS. Compared with the normal control group,percentages of CD3^＋and CD3^＋CD4^＋were lower inPSYDS（ P 〈0. 05）. The ratio of CD3^＋CD4^＋/ CD3^＋CD8＋was lower in GGDHS and PSYDS than in the normal control group（ P 〈0. 05）. There was no statistical difference in the CD3^＋CD8^＋percentage among various syndrome types（ P 〈0. 05）. The quantitation of HLA-DR13 gene was lower in GDPDS and GSYDS than in the normal control group（ P 〈0. 05）. The positive rate of BCP mutation was higher in GSYDS than in other syndrome types（ P 〈0. 05）. Conclusion Co-detection results of HLA-DR13 and BCP could be used as reference indices of Chinese medical syndrome typing of CHB.