HBV相关慢加急性肝衰竭患者血清中miR-122的表达及其临床意义

发布时间：2018-04-19 18:48

本文选题：miR-122 + HBV　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:分析HBV相关慢加急性肝衰竭(HBV-ACLF)患者血清中miR-122表达水平与各临床指标的相关性,进一步探讨血清中miR-122在HBV-ACLF疾病评估方面的潜在价值。方法:1.连续收集山西医科大学第一医院感染病科就诊的HBV-ACLF患者血清样本49例及同期慢性乙型肝炎(CHB)患者血清样本30例,并通过体检中心收集同期健康体检者(HC)的血清样本30例,分别设为HBV-ACLF组、CHB组、HC组(正常对照组),并根据临床表现的严重程度将HBV-ACLF患者分为早期、中期、晚期3组。2.使用RNAiso PLUS Total RNA提取试剂提取各血清标本中总RNA。通过miRNA RT-PCR试剂盒对所提取的总RNA进行目的基因及内参基因的反转录,并进一步行实时荧光定量PCR检测血清样本miR-122的相对表达水平。本实验以snRNAu6为内参,miR-122相对表达量用2-ΔCT表示。结果采用SPSS 22.0进行统计分析。3.比较不同组间miR-122表达水平的差异并分析HBV-ACLF患者血清中miR-122的表达与各生化指标的相关性。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,从而间接反映miR-122水平与疾病严重程度的关系。采用受试者工作特征曲线(ROC曲线)分析血清中miR-122在HBV-ACLF诊断评估方面的潜在价值。结果:1.HBV相关慢加急性肝衰竭患者、慢性乙型肝炎患者、健康体检者患者3组研究对象样本血清中miR-122的相对表达量分别为:HBV-ACLF组51.27(29.35-82.73)、CHB组15.35(11.49-22.43)、HC组8.03(5.87-8.88)。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(P0.001)。HBV相关慢加急性肝衰竭患者不同临床分期样本血清中miR-122的相对表达量分别为:早期32.45(17.54-45.73)、中期66.17(47.24-99.46)、晚期85.63(81.01-146.02),在晚期HBV-ACLF患者血清中miR-122的表达水平较早期及中期上调明显(P0.001、P=0.048)。2.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,P0.001;与PTA呈负相关,r=-0.797,P0.001。非条件Logistic回归分析结果显示,对于早期而言,血清中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%(OR 95%CI:1.013~1.104),进展为晚期的风险增加13.0%(OR 95%CI:1.034~1.235)。ROC分析显示,miR-122评价疾病诊断的曲线下面积(AUC)为0.942(95%CI,0.903-0.982),灵敏度:83.7%,特异度:86.7%。结论:血清中miR-122的表达水平可作为评价HBV-ACLF疾病严重程度的潜在生物学标志物,并在HBV-ACLF疾病诊断评估方面具有一定的参考价值。
[Abstract]:Objective: to analyze the correlation between serum miR-122 expression and clinical indexes in patients with chronic and acute liver failure (HBV), and to explore the potential value of miR-122 in the evaluation of HBV-ACLF disease. Method 1: 1. The serum samples of 49 patients with HBV-ACLF and 30 patients with chronic hepatitis B were collected continuously from the infectious department of the first Hospital of Shanxi Medical University. The patients with HBV-ACLF were divided into three groups according to the severity of clinical manifestations: early, middle and late stages. RNAiso PLUS Total RNA extraction reagent was used to extract total RNA from each serum sample. MiRNA RT-PCR kit was used to reverse transcription of target gene and internal reference gene of total RNA, and real-time quantitative PCR was used to detect the relative expression level of miR-122 in serum samples. In this experiment, the relative expression of miR-122 was expressed by 2-螖 CT using snRNAu6 as the internal reference. Results SPSS 22.0 was used for statistical analysis. To compare the difference of miR-122 expression among different groups and to analyze the correlation between the expression of miR-122 in serum of HBV-ACLF patients and the biochemical indexes. Furthermore, the role of miR-122 in the progression of HBV-ACLF was analyzed by non-conditional Logistic regression analysis, which indirectly reflected the relationship between the level of miR-122 and the severity of the disease. The potential value of serum miR-122 in the diagnosis and evaluation of HBV-ACLF was analyzed by using the operating characteristic curve (ROC curve). Results: 1. The relative expression of miR-122 in the serum of the patients with chronic hepatitis B, chronic hepatitis B and healthy people were 51.2729.35-82.73CHB 15.35, 11.49-22.43C and 8.035.87-8.88, respectively. The relative expression of miR-122 in the serum of the patients with chronic hepatitis B, chronic hepatitis B and healthy controls were 5.037-8.888.87-8.87-8.87-8.87-8.87-8.87-8.87-8.87-8.88 respectively. Compared with the serum miR-122 expression in healthy controls and CHB patients, the expression of miR-122 in serum of HBV-ACLF patients was up-regulated (P 0.001). The relative levels of miR-122 expression in serum of patients with chronic HBV-associated chronic and acute hepatic failure were as follows: Phase 32.45 ~ 17.54-45.73, middle stage 66.17 ~ 47.24-99.46, late stage 85.63 ~ 81.01-146.02, the expression level of miR-122 in serum of patients with advanced HBV-ACLF was significantly higher than that of early and middle stage. 2. The expression of miR-122 in serum of patients with HBV-ACLF was significantly higher than that of patients with advanced HBV-ACLF. There was a positive correlation between TBIL expression and serum total bilirubin level (P 0.001), and a negative correlation with PTA (P 0.001). The results of non-conditional Logistic regression analysis showed that in early stage, the relative expression of miR-122 in serum was 2- 螖 CT. The risk of HBV-ACLF progression was increased by 5.8%(OR 95: 1.013 1.104 and advanced stage. 13.0%(OR 95%CI:1.034~1.235).ROC analysis showed that the area under the curve for evaluating the diagnosis of HBV-ACLF was 0.942% 95% CII 0.903-0.982C, sensitivity 83.7%, and specificity 86.7%. Conclusion: the expression of miR-122 in serum can be used as a potential biomarker to evaluate the severity of HBV-ACLF disease, and it has some reference value in the diagnosis and evaluation of HBV-ACLF disease.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R512.62;R575.3

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