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细胞Toll样受体4/微小核糖核酸-181a在慢性乙型肝炎肝纤维化进程中的动态表达及临床意义

Dynamic expressions and clinical significance of toll-like receptor 4/microRNA-181a in the pathogenesis of hepatic fibrosis in patients with chronic hepatitis B

摘要:

目的 探讨TLR4/微小核糖核酸(micro RNA,miRNA)-181a在CHB肝纤维化进程中的作用及临床意义.方法 CHB患者经肝活组织检查进行肝纤维化分期(S);实时PCR法检测肝组织、血清miRNA-181a以及肝组织TLR4 mRNA表达;蛋白质印迹法检测肝组织TLR4蛋白;计算基于4因子的纤维化指数(fibrosis index based on the 4 factors,FIB-4)及天冬氨酸转氨酶与血小板比值指数(aspartate aminotransferase-to-platelet ratio index,APRI)进行非侵袭性肝纤维化诊断;对数据采用单因素方差分析、Mann-Whitney U检验、Spearman相关分析及受试者工作特征曲线(receiver operation characteristic curve,ROC曲线)分析.结果 40例CHB患者,S0期(7例)、S1期(6例)、S2期(14例)、S3期(7例)和S4期(6例)血清miRNA-181a表达水平(2-ΔΔCt值)分别为1.00±0.00、0.68±0.08、1.60±0.43、2.32±0.40和1.81±0.22,总体呈上升趋势(F=207.242,P=0.000),与肝纤维化严重程度呈正相关(r=0.754,P<0.01);随肝纤维化分期加重,肝组织miRNA-181a、TLR4 mRNA及TLR4蛋白表达总体呈上升趋势(F值分别为207.242,110.390和57.030,均P<0.01),肝组织miRNA-181a与T L R4蛋白表达及肝纤维化严重程度均呈正相关(r值分别为0.673和0.911,均P<0.01).严重肝纤维化(S3~S4)组APRI与非严重肝纤维化(S0~S2)组差异无统计学意义(Z=-1.401,P>0.05).血清miRNA-181a表达评估严重肝纤维化(S3~S4)优于FIB-4(ROC曲线下面积:0.887比0.695;准确度:85.0% 比60.0%).结论 miRNA-181a可能参与TLR4信号通路的调控而影响CHB患者肝纤维化进程,有望成为防治肝纤维化新的"靶点"及非侵袭性评估肝纤维化的血清指标.

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abstracts:

Objective To explore the dynamic expressions and clinical significance of toll-like receptor 4 (TLR4)/microRNA (miRNA)-181a in the pathogenesis of hepatic fibrosis (HF) in patients with chronic hepatitis B (CHB) .Methods CHB patients underwent liver biopsy for fibrosis staging HF (S) .Real-time polymerase chain reaction (PCR) was used to detect the expressions of miRNA-181a in both serum and liver tissue and the expression of TLR4 mRNA in liver tissue .Western blot was used to detect the expression of TLR4 protein in liver tissue . The fibrosis-4 (FIB-4 ) index and aspartate aminotransferase-to-platelet ratio index (APRI ) were calculated for noninvasive evaluation of fibrosis staging .One-way ANOVA ,Mann-Whitney U test ,spearman correlation analysis and receiver operating characteristic (ROC) curve were used for statistical analysis .Results Forty CHB patients were includedin this study ,including 7 with S0 ,6 with S1 ,14 with S2 ,7 with S3 and 6 with S4 .Serum levesl of miRNA-181a (2-ΔΔCt ) in paitents with S0-4 were 1 .00 ± 0 .00 ,0 .68 ± 0 .08 ,1 .60 ± 0 .43 ,2 .32 ± 0 .40 , and 1 .81 ± 0 .22 ,respectively ,showing an overall upward trend (F=207 .242 ,P< 0 .01) and a positive correlation with the severity of HF (r= 0 .754 , P< 0 .01) .The expressions of miRNA-181a ,TLR4 mRNA and TLR4 protein in liver tissues showed an overall increasing trend from S 0 to S4 (F=207 .242 , 110 .390 and 57 .030 ,respectively ,all P<0 .01) .The expression of miRNA-181a in liver tissue showed a positive correlation with both the expression of TLR4 protein in liver tissue and the severity of HF (r=0 .673 and 0 .911 ,respectively ,both P< 0 .01) .There was no significant difference of APRI scores between the severe (S3-4) and non-severe (S0-2) HF groups (Z= -1 .401 ,P>0 .05) .The serum level of miRNA-181a was superior to FIB-4 index for evaluation of the severe HF (S3-4) ,with areas under the ROC curve (AUROC ) of 0 .887 and 0 .695 , respectively , and accuracy of 85 .0% and 60 .0% , respectively .Conclusions miRNA-181a may be involved in the regulation of TLR4 signaling pathway so that to affect the progression of HF in CHB patients ,which may be a potential new target for the prevention and early treatment of HF and a non-invasive serum marker for evaluation of HF .

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作者: 陈靖 [1] 郑琦 [1] 曾达武 [1] 陈薇 [1] 朱月永 [1]
期刊: 《中华传染病杂志》2018年36卷8期 466-472页 ISTICCSCD
栏目名称: 论著
DOI: 10.3760/cma.j.issn.1000-6680.2018.08.003
发布时间: 2018-11-05
基金项目:
国家科技重大专项项目 福建省自然科学基金项目 福建省卫生系统中青年骨干人才培养项目(2015-ZQN-JC-25)National Science and Technology Major Project The Natural Science Foundation of Fujian Province Training Project for Young and Middle-Aged Core Talents of Health System of Fujian Province
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