慢性乙肝病毒感染者I型干扰素受体与机体氧化应激状态的相关性研究

发布时间：2018-02-27 02:25

本文关键词： 乙型肝炎慢性干扰素 Ⅰ型氧化损伤　出处：《山东大学》2013年硕士论文　论文类型：学位论文

【摘要】：研究背景与目的干扰素-α是目前针对慢性乙型肝炎(CHB)的一线抗病毒药物。它本身不具备直接杀伤、抑制乙肝病毒(HBV)的能力,其抗病毒作用的发挥有赖于它与细胞膜表面Ⅰ型干扰素受体(IFNAR)的结合,生成抗病毒蛋白,抑制HBV复制。IFNAR参与CHB的病程进展,而氧化应激则参与CHB的发病过程,同时可影响包括I FNAR在内的蛋白质的结构与功能。本研究通过测定慢性HBV感染者机体外周血IFNAR的表达水平及机体氧化损伤与抗氧化损伤相关指标的水平,探讨患者外周血IFNAR与机体氧化应激状态的关系,进一步揭示CHB的发病机制并对CHB抗病毒治疗过程中IFN的临床应用提供指导。研究方法 CHB患者54例,乙肝后肝硬化(失代偿期)患者31例,所有患者符合2010年《慢性乙型肝炎防治指南》中的诊断标准。11例健康志愿者为正常对照。外周血淋巴细胞和单核细胞内Ⅰ型干扰素受体1(IFNAR1)和Ⅰ型干扰素受体2(IFNAR2)的水平经由流式细胞仪技术(FCM)测定。IFNARl和Ⅰ型干扰素受体2c(IFNAR2c)mRNA在外周血单个核细胞(PBMCs)中的表达情况经由实时荧光定量PCR法(RT-PCR)测定。血浆可溶性干扰素α//β受体(soluble IFNAR)水平由酶联免疫吸附法(ELISA)测定。衡量机体氧化损伤程度的指标--黄嘌呤氧化酶(XOD),丙二醛(MDA),谷胱甘肽(GSH),谷胱甘肽巯基转移酶(GST)和谷胱甘肽过氧化物酶(GSH-Px)在血浆中的水平经由ELISA测定。结果 1.CHB和肝硬化患者单核细胞及淋巴细胞内IFNAR1和IFNAR2的水平显著高于正常对照组。CHB和肝硬化患者PBMCs中IFNARl和IFNAR2c mRNA表达显著高于正常对照组。CHB和肝硬化患者血浆可溶性IFNAR水平较正常对照组明显上调。其中,单核细胞中IFNAR2的平均荧光强度(MFI)在CHB患者中显著高于肝硬化患者,差异具有统计学意义。CHB患者中,外周血IFNAR2阳性淋巴细胞率与血清HBV DNA存在正相关联系(r=0.363,p=0.041),而外周血淋巴细胞中IFNAR1的MFI与血清HBV DNA表现为负相关(r=-0.355,p=0.027)。 2.CHB及肝硬化患者血浆XOD,MDA和GST水平较正常对照组明显上调,而抗氧化损伤指标如GSH,GSH-Px水平则CHR及肝硬化组明显低于正常对照组。此外,CHB患者血浆MDA, GSH和GST水平显著高于肝硬化患者 3.CHB组中,血浆XOD水平与血清总胆红素(TBIL)水平呈显著正相关(r=0.255,p=0.044)。血浆MDA水平与血清ALT、GGT水平呈正相关(r=0.410,p=0.001；r=0.488,p0.001)。血浆GSH水平与血清TBIL。水平呈负相关(r=-0.394,p=0.009)。血浆GST水平与血清ALT水平呈正相关(r=0.320,p=0.008)。 4.CHB组中,血浆GST水平与淋巴细胞中IFNAR2的MFI呈明显负相关(r=-0.447,p=0.01)。结论慢性乙肝病毒感染者外周淋巴细胞和单核细胞内Ⅰ型干扰素受体的表达显著上调。同时,慢性乙肝病毒感染者体内存在氧化应激。慢性乙型肝炎患者外周血淋巴细胞内IFNAR2的平均荧光强度与血浆GST水平的负相关趋势提示氧化应激对慢性乙型肝炎患者外周血IFNAR的表达起重要调节作用。
[Abstract]:Research background and purpose
Interferon alpha is present in chronic hepatitis B (CHB) first-line antiviral drugs. It does not have direct cytotoxicity, inhibition of hepatitis B virus (HBV) the ability to play its antiviral action depends on the surface of the cell membrane and its type of interferon receptor (IFNAR) binding to antiviral protein, inhibit the progression of HBV copy the.IFNAR CHB to participate in the pathogenesis of oxidative stress is involved in CHB, at the same time can affect the structure and function of proteins including I, FNAR. The expression level of related indexes and oxidative damage and antioxidant determination in chronic HBV infection in peripheral blood IFNAR damage level, to explore the relationship between peripheral blood of patients with IFNAR and oxidative stress, further reveal the pathogenesis of CHB and provide guidance for clinical application of CHB in the process of antiviral treatment of IFN.
research method
54 cases of CHB patients, liver cirrhosis (decompensated) in 31 patients, all patients met the diagnostic criteria of.11 cases of healthy volunteers in 2010 guidelines for prevention and treatment of chronic hepatitis B in the < > as normal control group. Peripheral blood lymphocytes and mononuclear cells in type I interferon receptor 1 (IFNAR1) and 2 (type I interferon receptor the level of IFNAR2) by flow cytometry (FCM) determination of.IFNARl and type I interferon receptor 2C (IFNAR2c) mRNA in peripheral blood mononuclear cells (PBMCs) expression in the real time fluorescence quantitative PCR (RT-PCR) assay. Plasma soluble interferon alpha / beta receptor (soluble IFNAR) levels by enzyme linked immunosorbent assay (ELISA) determination of xanthine oxidase index to measure the degree of oxidative damage (XOD), malondialdehyde (MDA), glutathione (GSH), glutathione S-transferase (GST) and glutathione peroxidase (GSH-Px) levels in plasma Measured by ELISA.
Result
Mononuclear cells and lymphocytes in patients with liver cirrhosis and 1.CHB in IFNAR1 and IFNAR2 levels were significantly higher than those in normal control group.CHB and PBMCs in patients with liver cirrhosis in the expression of IFNARl and IFNAR2c mRNA were significantly higher than that of normal control group in patients with.CHB and liver cirrhosis soluble IFNAR levels compared with normal control group increased significantly. The average fluorescence intensity of IFNAR2 in monocytes (MFI) was significantly higher than that in patients with liver cirrhosis in patients with CHB, the difference was statistically significant in patients with.CHB, there is a positive correlation between peripheral blood lymphocytes and serum IFNAR2 positive rate of HBV DNA (r=0.363, p=0.041), MFI DNA IFNAR1 and serum HBV in peripheral blood lymphocytes showed a negative correlation (r=-0.355, p=0.027).
The levels of plasma XOD, MDA and GST in 2.CHB and cirrhosis patients were significantly higher than those in the normal control group, while the antioxidant injury indicators such as GSH and GSH-Px levels were significantly lower in the CHR and cirrhosis group than those in the normal control group. In addition, the plasma MDA, GSH and GST levels in CHB patients were significantly higher than those in cirrhosis patients.
In the 3.CHB group, the level of plasma XOD and serum total bilirubin (TBIL) levels were positively correlated (r=0.255, p=0.044). The level of plasma MDA and serum ALT, GGT levels were positively correlated (r=0.410, p=0.001; r=0.488, p0.001). The plasma GSH level was negatively correlated with serum TBIL. levels (r=, -0.394, p=0.009). GST level was positively with the serum level of ALT (r=0.320, p=0.008).
In the group 4.CHB, the level of plasma GST was negatively correlated with the MFI of IFNAR2 in the lymphocyte (r=-0.447, p=0.01).
conclusion
Significantly increased expression of chronic hepatitis B virus infection in peripheral lymphocytes and mononuclear cells in type I interferon receptor. At the same time, chronic hepatitis B virus infection in the presence of oxidative stress. A negative trend in patients with chronic hepatitis B in peripheral blood lymphocytes in IFNAR2 average fluorescence intensity and plasma GST level suggesting that oxidative stress of patients with chronic hepatitis B the peripheral blood IFNAR expression play an important role.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R512.62

【共引文献】