CAJ | 학술논문

目的探讨微RNA-548ah (miR-548ah)和微RNA-4804 (miR-4804)在慢性HBV感染不同阶段的患者外周血单核细胞(PBMC)中的表达及临床意义.方法选取南通大学附属泰州市人民医院2012年9月至2013年8月住院和门诊HBV感染者85例,其中慢性乙型肝炎(CHB)患者29例,慢性HBV携带者(HBVC) 30例,非活动性HBsAg携带者(IASC) 26例.另选同期健康体检者28名作为对照.采用反转录-实时荧光定量PCR (qRT-PCR)技术检测PBMC中miR-548ah和miR-4804的表达.miRNA分子诊断HBV感染免疫耐受期和清除期的价值采用受试者工作特征(ROC)曲线分析,微RNA (miRNA)分子与临床主要指标丙氨酸转氨酶(ALT)和HBV DNA的相关性采用Pearson分析.结果 miR-548ah和miR-4804在CHB组、HBVC组、IASC组和健康对照组患者PBMC中的表达差异有统计学意义(F=28.16和83.17,P＜0.01).与健康对照组比较,miR-548ah的表达在CHB组明显上调(P＜0.01),在HBVC及IASC组明显下调(P＜0.01);miR-4804的表达在CHB组明显上调(P＜0.01),在HBVC组明显下调(P＜0.01),但在IASC组的表达与健康对照组比较差异无统计学意义(P＞0.05).miR-548ah和miR-4804区分慢性HBV感染免疫耐受期和清除期的ROC曲线下面积(AUC)分别为0.966和0.997,敏感度分别为89.7％和96.6％,特异度分别为99.6％和100.0％.miR-548ah和miR-4804的表达水平与ALT和HBV DNA水平均无相关性(r=0.14,0.18,-0.20和-0.19,P＞0.05).结论 miR-548ah和miR-4804的异常表达可能与CHB的免疫发病机制有关,且有助于区分CHB的免疫耐受期和清除期.
목적 탐토미RNA-548ah (miR-548ah)화미RNA-4804 (miR-4804)재만성HBV감염불동계단적환자외주혈단핵세포(PBMC)중적표체급림상의의.방법 선취남통대학부속태주시인민의원2012년9월지2013년8월주원화문진HBV감염자85례,기중만성을형간염(CHB)환자29례,만성HBV휴대자(HBVC) 30례,비활동성HBsAg휴대자(IASC) 26례.령선동기건강체검자28명작위대조.채용반전록-실시형광정량PCR (qRT-PCR)기술검측PBMC중miR-548ah화miR-4804적표체.miRNA분자진단HBV감염면역내수기화청제기적개치채용수시자공작특정(ROC)곡선분석,미RNA (miRNA)분자여림상주요지표병안산전안매(ALT)화HBV DNA적상관성채용Pearson분석.결과 miR-548ah화miR-4804재CHB조、HBVC조、IASC조화건강대조조환자PBMC중적표체차이유통계학의의(F=28.16화83.17,P＜0.01).여건강대조조비교,miR-548ah적표체재CHB조명현상조(P＜0.01),재HBVC급IASC조명현하조(P＜0.01);miR-4804적표체재CHB조명현상조(P＜0.01),재HBVC조명현하조(P＜0.01),단재IASC조적표체여건강대조조비교차이무통계학의의(P＞0.05).miR-548ah화miR-4804구분만성HBV감염면역내수기화청제기적ROC곡선하면적(AUC)분별위0.966화0.997,민감도분별위89.7％화96.6％,특이도분별위99.6％화100.0％.miR-548ah화miR-4804적표체수평여ALT화HBV DNA수평균무상관성(r=0.14,0.18,-0.20화-0.19,P＞0.05).결론 miR-548ah화miR-4804적이상표체가능여CHB적면역발병궤제유관,차유조우구분CHB적면역내수기화청제기.
Objective To investigate the expression of microRNA-548ah (miR-548ah) and microRNA-4804 (miR-4804) in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis B virus infection and their clinical significance.Methods PBMCs were collected from 29 patients with chronic hepatitis B (CHB),30 hepatitis B virus carriers (HBVC),26 inactive HBsAg carriers (IASC) and 28 healthy controls in Taizhou People's Hospital during September 2012 and August 2013.Expressions of miR-548ah and miR-4804 in PBMCs were detected by fluorescence real-time quantitative RT-PCR (qRT-PCR).Receiver operating characteristic (ROC) curve was used to evaluate the expression of miR-548ah and miR-4804 in distinguishing immune tolerance phase and clearance phase of HBV infection.Pearson correlation analysis was performed to assess the correlations of miRNAs expression with clinical markers alanine aminotrans ferase (ALT) and HBV DNA loads.Results There were significant differences in expressions of miR-548ah and miR-4804 in PBMCs between CHB,HBVC,IASC groups and control group (F =28.16 and 83.17,P ＜ 0.01).Compared with control group,miR-548ah was up-regulated in CHB group(P ＜ 0.01),and down-regulated in HBVC and IASC groups (P ＜ 0.01) ; miR-4804 was up-regulated in CHB group (P ＜ 0.01),down-regulated in HBVC group (P ＜ 0.01),while there was no significant difference between IASC group and control group in miR-4804 expressions(P ＞ 0.05).The areas under ROC curve (AUCs) of miR-548ah and miR4804 in differentiation of immune tolerance and immune clearance were 0.966 and 0.997,and the sensitivities and specificities were 89.7％,96.6％ and 99.6％,100.0％,respectively.No significant correlation was found between the expression of miR-548ah,miR-4804 and ALT,HBV DNA loads (r=0.14,0.18,-0.20 and-0.19,P＞0.05).Conclusion miR-548ah and miR-4804 may be involved in the immunopathogenesis of CHB,and their expression levels in PBMCs are helpful in differentiation of immune tolerance and immune clearance in HBV infection.