慢加急性肝衰竭患者外周血肿瘤坏死因子-α基因启动子甲基化状态的研究  

Hypomethylation of TNF-α gene promoter in the patients with acute-on-chronic hepatitis B liver failure

作　　者：韩利岩[1] 郭朝阳[1] 范玉琛[1] 王凯[1] 

机构地区：[1]山东大学齐鲁医院肝病科,济南250012

出　　处：《中华实验和临床病毒学杂志》2011年第5期368-370,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：基金项目：国家十一五重大专项（2008ZX10002-007）

摘　　要：目的探讨肿瘤坏死因子-仅及表观遗传调控在慢加急性肝衰竭发病机制中的可能作用。方法ELISA法检测外周血TNF-α的表达，并与患者的肝功能进行相关性分析。同时提取外周血单个核细胞DNA，重亚硫酸盐处理后，用针对改变后的DNA序列设计特异性引物并进行聚合酶链反应（PCR）。根据修饰后DNA序列的不同，运用在线MethPrimer软件设计引物，甲基化特异性聚合酶链反应（Methylationspecificpolymerasechainreaction，MSP）检测标本中TNF-α扩增产物，分析TNF-α启动子甲基化与血清TNF-α表达的相关性，并分析甲基化与病情严重程度的相关性。采用SPSS16．0软件进行统计分析。结果慢加急性肝衰竭患者组外周血TNF-α蛋白水平（44．9260±26．48523）均高于慢性乙型病毒性肝炎患者组（18．92505±9．04461）和健康对照组（11．9172±5．04612），且各组间比较差异均具有统计学意义（P〈0．05）。慢加急性肝衰竭患者组外周血清TNF—a水平与血清TBIL及终末期肝脏病模型（MELD）评分之间呈明显的正相关，与PTA呈明显的负相关。慢加急性肝衰竭组、慢性乙型病毒性肝炎患者组、健康对照组TNF-α启动子甲基化状态的差别有统计学意义。慢加急性肝衰竭患者与慢性乙型病毒性肝炎患者及健康对照甲基化组血清TNF-α水平均明显低于非甲基化组（P〈0．05）。结论TNF-α与肝炎活动及肝细胞损害有密切关系，DNA甲基化在慢加急肝衰竭患者中参与调控细胞因子的表达中。Objective The present study was designed to investigate the possible epigenetic alteration in the promoter of TNF-α in the patients with acute-on-chronic hepatitis B liver failure （ACHBLF）. Methods The methylation of TNF-α promoter in peripheral blood mononuclear cells （PBMCs） was measured by methylation specific PCR （MSP）. The level of serum TNF-α was determined by enzyme-linked immunosorbent assay （ELISA）. Model for End-stage Liver Disease （MELD） was performed for the evaluation of liver failure. Results The serum level of TNF-ct in patients with ACHBLF（44. 9260 ± 26.48523 ） was higher than that in C HB （ 18. 92505 ± 9.04461 ） and healthy controls （ 11. 9172 ± 5. 04612 ） （P 〈 0. 05 ）. Moreover, the serum TNF-α level was significantly decreased in methylation group as compared to unmethylaiton group in patients with ACHBLF（P 〈 0. 05 ）. MELD was not significantly different between methylated and unmethylated group of ACHBLF patients （ P 〉 0.05 ）. In addition, the serum level of TNF-α was found to be positively correlated with serum total bilirubin （r = 0. 891 ,P 〈 0.01 ）and MELD seore（r = 0. 792 ,P 〈 0. 01 ）, but to be negatively correlated with prothrombin activity （ r = - 0. 511, P 〈 0. 05 ） in patients with ACHBLF. Conclusion The TNF-α methylation patten is stable for the liver failure, suggesting the effect of environment on methylation.

关 键 词：肝功能衰竭 肿瘤坏死因子 DNA 聚合酶链反应 

分 类 号：R575.3[医药卫生—消化系统]