CAJ | 학술논문

目的探讨肿瘤坏死因子-α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(Methylation specific polymerase chain reaction,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-α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.
목적 탐토종류배사인자-α급표관유전조공재만가급성간쇠갈발병궤제중적가능작용.방법 ELISA법검측외주혈TNF-α적표체,병여환자적간공능진행상관성분석.동시제취외주혈단개핵세포DNA,중아류산염처리후,용침대개변후적DNA서렬설계특이성인물병진행취합매련반응(PCR).근거수식후DNA서렬적불동,운용재선MethPrimer연건설계인물,갑기화특이성취합매련반응(Methylation specific polymerase chain reaction,MSP)검측표본중TNF-α확증산물,분석TNF-α계동자갑기화여혈청TNF-α표체적상관성,병분석갑기화여병정엄중정도적상관성.채용SPSS16.0연건진행통계분석.결과 만가급성간쇠갈환자조외주혈TNF-α단백수평(44.9260±26.48523)균고우만성을형병독성간염환자조(18.92505±9.04461)화건강대조조(11.9172±5.04612),차각조간비교차이균구유통계학의의(P＜0.05).만가급성간쇠갈환자조외주혈청TNF-α수평여혈청TBIL급종말기간장병모형(MELD)평분지간정명현적정상관,여PTA정명현적부상관.만가급성간쇠갈조、만성을형병독성간염환자조、건강대조조TNF-α계동자갑기화상태적차별유통계학의의.만가급성간쇠갈환자여만성을형병독성간염환자급건강대조갑기화조혈청TNF-α수평균명현저우비갑기화조(P＜0.05).결론 TNF-α여간염활동급간세포손해유밀절관계,DNA갑기화재만가급간쇠갈환자중삼여조공세포인자적표체중.
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-α in patients with ACHBLF(44.9260 ±26.48523 ) was higher than that in CHB ( 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-αwas found to be positively correlated with serum total bilirubin ( r =0.891,P ＜ 0.01 ) and MELD score ( 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.