乙型肝炎相关慢加急性肝功能衰竭患者血清中miR-122的表达及其临床意义

doi:10.3877/cma.j.issn.1674-1358.2017.06.004

目的

分析乙型肝炎（HBV）病毒感染相关慢加急性肝功能衰竭（HBV-ACLF）患者血清中miR-122表达水平与各临床指标是否有相关性，从而探讨血清中miR-122在评价HBV-ACLF疾病严重程度、诊断中的价值。

方法

收集山西医科大学第一医院就诊的HBV-ACLF患者血清样本49例、慢性乙型肝炎（CHB）患者血清样本30例及健康体检者的血清样本30例，通过实时荧光定量PCR检测各个样本miR-122的相对表达水平。以u6为内参，miR-122相对表达量用2^-ΔCT表示。根据临床表现的严重程度将HBV-ACLF患者分为早期、中期和晚期组，比较不同组间miR-122表达水平的差异及其与生化指标的相关性。进一步通过非条件Logistic回归分析miR-122在HBV-ACLF病程进展中的作用，从而反映miR-122与疾病严重程度的关系。采用受试者工作特征曲线（ROC）评价血清中miR-122在HBV-ACLF诊断评估方面的价值。

结果

与健康体检者及CHB患者血清中miR-122的表达水平相比，HBV-ACLF患者血清中miR-122的表达水平上调（P < 0.001），且在晚期HBV-ACLF患者血清中miR-122的表达水平较早期及中期显著上调（P < 0.001、P = 0.048）。HBV-ACLF患者血清miR-122表达水平的升高，与血清中总胆红素（TBil）的表达水平呈正相关（r = 0.508、P < 0.001）；与PTA呈负相关（r =-0.797、P < 0.001）。非条件Logistic回归分析结果显示对于HBV-ACLF早期而言，miR-122相对表达值2^-ΔCT每增加一个单位，引起HBV-ACLF进展为中期的风险增加5.8%（95%CI：1.013～1.104），进展为晚期的风险增加13.0%（95%CI：1.034～1.235）。ROC分析结果显示，miR-122评价疾病的曲线下面积（AUC）为0.942（95%CI：0.903～0.982），灵敏度为83.7%，特异度为86.7%。

结论

血清中miR-122的表达水平可成为评价HBV-ACLF的严重程度及作为HBV-ACLF诊断的潜在生物标志。

关键词: miR-122, 肝炎病毒、乙型, 肝功能衰竭、慢加急性, 肝炎、乙型、慢性

Objective

To investigate the relationship between the expression of miR-122 in HBV-related acute-on-chronic liver failure (HBV-ACLF) and clinical indicators, and to analyze the role of miR-122 in evaluating the severity and diagnosis of HBV-ACLF.

Methods

Serum samples from 49 patients with HBV-ACLF, 30 patients with chronic hepatitis B (CHB) and 30 healthy volunteers were collected from the First Affiated Hospital of Shanxi Medical University. The relative expression of miR-122 in each sample was detected by fluorescence real-time quantitative PCR. With the u6 as endogenous control, the relative expression of miR-122 was expressed as 2^-ΔCT. Patients with HBV-ACLF were divided into early stage, middle stage and late stage groups according to the severity of clinical manifestations. The levels of miR-122 expression between different groups were compared, and the correlation between the expression of miR-122 and biochemical markers were analyzed, respectively. Furthermore, the role of miR-122 in the progression of HBV-ACLF was analyzed by Unconditional Logistic regression model, thus reflecting the association between miR-122 and disease severity. Receiver operating characteristic curve (ROC) was used to evaluate the value of serum miR-122 in the diagnosis of HBV-ACLF.

Results

The expression level of miR-122 in serum of patients with HBV-ACLF was significantly higher than those of healthy subjects and patients with CHB (P < 0.001), and the level of miR-122 expression in advanced HBV-ACLF was significantly higher than those of the early and middle stages (P < 0.001, P = 0.048). The serum level of miR-122 in patients with HBV-ACLF was positively correlated with the level of serum total bilirubin (TBil) (r = 0.508, P < 0.001) and negatively correlated with PTA (r =-0.797, P < 0.001). Unconditional Logistic regression model showed that an increase of one unit of relative expression of miR-122 (2^-ΔCT) was associated with 5.8% (95%CI: 1.013-1.104) increase in the risk of progression from HBV-ACLF early stage to middle stage and 13.0% (95%CI: 1.034-1.235) increase in the risk from early stage to late stage. The ROC analysis showed that the area under the curve (AUC) of disease diagnosis was 0.942 (95%CI: 0.903-0.982), the sensitivity and the specificity were 83.7% and 86.7%.

Conclusions

The expression level of miR-122 in serum could be used as a biomarker for the diagnosis and the severity of HBV-ACLF.

Key words: miRNA -122 (miR-122), Hepatitis B virus (HBV), Acute-on-chronic liver failure, Chronic hepatitis B

图1 各组患者血清miR-122的相对表达水平

图2 HBV-ACLF患者血清中miR-122的表达与血清TBil、PTA的相关性

表1 影响早期HBV-ACLF病情进展的非条件Logistic回归分析

变量^a		b值	Wald χ²值	P值	OR值	95%CI
中期
	年龄	0.081	1.325	0.250	1.085	0.944～1.246
	BMI	0.048	0.035	0.851	1.049	0.639～1.720
	2^-ΔCT	0.056	6.472	0.011	1.058	1.013～1.104
	胆红素	0.011	3.692	0.055	1.011	1.000～1.023
	性别	-0.335	0.065	0.798	0.715	0.055～9.343
晚期
	年龄	0.079	0.289	0.591	1.082	0.811～1.443
	BMI	0.470	1.533	0.216	1.600	0.760～3.367
	2^-ΔCT	0.122	7.236	0.007	1.130	1.034～1.235
	胆红素	0.034	8.365	0.004	1.034	1.011～1.058
	性别	0.289	0.017	0.896	1.335	0.017～101.880

表1 影响早期HBV-ACLF病情进展的非条件Logistic回归分析

变量^a		b值	Wald χ²值	P值	OR值	95%CI
中期
	年龄	0.081	1.325	0.250	1.085	0.944～1.246
	BMI	0.048	0.035	0.851	1.049	0.639～1.720
	2^-ΔCT	0.056	6.472	0.011	1.058	1.013～1.104
	胆红素	0.011	3.692	0.055	1.011	1.000～1.023
	性别	-0.335	0.065	0.798	0.715	0.055～9.343
晚期
	年龄	0.079	0.289	0.591	1.082	0.811～1.443
	BMI	0.470	1.533	0.216	1.600	0.760～3.367
	2^-ΔCT	0.122	7.236	0.007	1.130	1.034～1.235
	胆红素	0.034	8.365	0.004	1.034	1.011～1.058
	性别	0.289	0.017	0.896	1.335	0.017～101.880

图3 血清中miR-122的表达水平用于诊断HBV-ACLF的ROC曲线分析

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Expression of miR-122 in serum of patients with HBV-related acute-on-chronic liver failure and the clinical significance

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