本文選題：miR-122 + HBV��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:分析HBV相關(guān)慢加急性肝衰竭(HBV-ACLF)患者血清中miR-122表達水平與各臨床指標(biāo)的相關(guān)性,進一步探討血清中miR-122在HBV-ACLF疾病評估方面的潛在價值。方法:1.連續(xù)收集山西醫(yī)科大學(xué)第一醫(yī)院感染病科就診的HBV-ACLF患者血清樣本49例及同期慢性乙型肝炎(CHB)患者血清樣本30例,并通過體檢中心收集同期健康體檢者(HC)的血清樣本30例,分別設(shè)為HBV-ACLF組、CHB組、HC組(正常對照組),并根據(jù)臨床表現(xiàn)的嚴(yán)重程度將HBV-ACLF患者分為早期、中期、晚期3組。2.使用RNAiso PLUS Total RNA提取試劑提取各血清標(biāo)本中總RNA。通過miRNA RT-PCR試劑盒對所提取的總RNA進行目的基因及內(nèi)參基因的反轉(zhuǎn)錄,并進一步行實時熒光定量PCR檢測血清樣本miR-122的相對表達水平。本實驗以snRNAu6為內(nèi)參,miR-122相對表達量用2-ΔCT表示。結(jié)果采用SPSS 22.0進行統(tǒng)計分析。3.比較不同組間miR-122表達水平的差異并分析HBV-ACLF患者血清中miR-122的表達與各生化指標(biāo)的相關(guān)性。進一步通過非條件Logistic回歸分析miR-122在HBV-ACLF病程進展中的作用,從而間接反映miR-122水平與疾病嚴(yán)重程度的關(guān)系。采用受試者工作特征曲線(ROC曲線)分析血清中miR-122在HBV-ACLF診斷評估方面的潛在價值。結(jié)果:1.HBV相關(guān)慢加急性肝衰竭患者、慢性乙型肝炎患者、健康體檢者患者3組研究對象樣本血清中miR-122的相對表達量分別為:HBV-ACLF組51.27(29.35-82.73)、CHB組15.35(11.49-22.43)、HC組8.03(5.87-8.88)。與健康體檢者及CHB患者血清中miR-122的表達水平相比,HBV-ACLF患者血清中miR-122的表達水平上調(diào)(P0.001)。HBV相關(guān)慢加急性肝衰竭患者不同臨床分期樣本血清中miR-122的相對表達量分別為: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的表達水平較早期及中期上調(diào)明顯(P0.001、P=0.048)。2.HBV-ACLF患者血清miR-122表達水平的升高,與血清中總膽紅素(TBIL)的表達水平呈正相關(guān),r=0.508,P0.001;與PTA呈負(fù)相關(guān),r=-0.797,P0.001。非條件Logistic回歸分析結(jié)果顯示,對于早期而言,血清中miR-122相對表達值2-ΔCT每增加一個單位,引起HBV-ACLF進展為中期的風(fēng)險增加5.8%(OR 95%CI:1.013~1.104),進展為晚期的風(fēng)險增加13.0%(OR 95%CI:1.034~1.235)。ROC分析顯示,miR-122評價疾病診斷的曲線下面積(AUC)為0.942(95%CI,0.903-0.982),靈敏度:83.7%,特異度:86.7%。結(jié)論:血清中miR-122的表達水平可作為評價HBV-ACLF疾病嚴(yán)重程度的潛在生物學(xué)標(biāo)志物,并在HBV-ACLF疾病診斷評估方面具有一定的參考價值。
[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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R512.62;R575.3

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本文編號：1774281