本文關(guān)鍵詞： 乙型肝炎 慢性 干擾素 Ⅰ型 氧化損傷　出處：《山東大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：研究背景與目的 干擾素-α是目前針對慢性乙型肝炎(CHB)的一線抗病毒藥物。它本身不具備直接殺傷、抑制乙肝病毒(HBV)的能力,其抗病毒作用的發(fā)揮有賴于它與細(xì)胞膜表面Ⅰ型干擾素受體(IFNAR)的結(jié)合,生成抗病毒蛋白,抑制HBV復(fù)制。IFNAR參與CHB的病程進(jìn)展,而氧化應(yīng)激則參與CHB的發(fā)病過程,同時可影響包括I FNAR在內(nèi)的蛋白質(zhì)的結(jié)構(gòu)與功能。本研究通過測定慢性HBV感染者機(jī)體外周血IFNAR的表達(dá)水平及機(jī)體氧化損傷與抗氧化損傷相關(guān)指標(biāo)的水平,探討患者外周血IFNAR與機(jī)體氧化應(yīng)激狀態(tài)的關(guān)系,進(jìn)一步揭示CHB的發(fā)病機(jī)制并對CHB抗病毒治療過程中IFN的臨床應(yīng)用提供指導(dǎo)。 研究方法 CHB患者54例,乙肝后肝硬化(失代償期)患者31例,所有患者符合2010年《慢性乙型肝炎防治指南》中的診斷標(biāo)準(zhǔn)。11例健康志愿者為正常對照。外周血淋巴細(xì)胞和單核細(xì)胞內(nèi)Ⅰ型干擾素受體1(IFNAR1)和Ⅰ型干擾素受體2(IFNAR2)的水平經(jīng)由流式細(xì)胞儀技術(shù)(FCM)測定。IFNARl和Ⅰ型干擾素受體2c(IFNAR2c)mRNA在外周血單個核細(xì)胞(PBMCs)中的表達(dá)情況經(jīng)由實時熒光定量PCR法(RT-PCR)測定。血漿可溶性干擾素α//β受體(soluble IFNAR)水平由酶聯(lián)免疫吸附法(ELISA)測定。衡量機(jī)體氧化損傷程度的指標(biāo)--黃嘌呤氧化酶(XOD),丙二醛(MDA),谷胱甘肽(GSH),谷胱甘肽巰基轉(zhuǎn)移酶(GST)和谷胱甘肽過氧化物酶(GSH-Px)在血漿中的水平經(jīng)由ELISA測定。 結(jié)果 1.CHB和肝硬化患者單核細(xì)胞及淋巴細(xì)胞內(nèi)IFNAR1和IFNAR2的水平顯著高于正常對照組。CHB和肝硬化患者PBMCs中IFNARl和IFNAR2c mRNA表達(dá)顯著高于正常對照組。CHB和肝硬化患者血漿可溶性IFNAR水平較正常對照組明顯上調(diào)。其中,單核細(xì)胞中IFNAR2的平均熒光強(qiáng)度(MFI)在CHB患者中顯著高于肝硬化患者,差異具有統(tǒng)計學(xué)意義。CHB患者中,外周血IFNAR2陽性淋巴細(xì)胞率與血清HBV DNA存在正相關(guān)聯(lián)系(r=0.363,p=0.041),而外周血淋巴細(xì)胞中IFNAR1的MFI與血清HBV DNA表現(xiàn)為負(fù)相關(guān)(r=-0.355,p=0.027)。 2.CHB及肝硬化患者血漿XOD,MDA和GST水平較正常對照組明顯上調(diào),而抗氧化損傷指標(biāo)如GSH,GSH-Px水平則CHR及肝硬化組明顯低于正常對照組。此外,CHB患者血漿MDA, GSH和GST水平顯著高于肝硬化患者 3.CHB組中,血漿XOD水平與血清總膽紅素(TBIL)水平呈顯著正相關(guān)(r=0.255,p=0.044)。血漿MDA水平與血清ALT、GGT水平呈正相關(guān)(r=0.410,p=0.001；r=0.488,p0.001)。血漿GSH水平與血清TBIL。水平呈負(fù)相關(guān)(r=-0.394,p=0.009)。血漿GST水平與血清ALT水平呈正相關(guān)(r=0.320,p=0.008)。 4.CHB組中,血漿GST水平與淋巴細(xì)胞中IFNAR2的MFI呈明顯負(fù)相關(guān)(r=-0.447,p=0.01)。 結(jié)論 慢性乙肝病毒感染者外周淋巴細(xì)胞和單核細(xì)胞內(nèi)Ⅰ型干擾素受體的表達(dá)顯著上調(diào)。同時,慢性乙肝病毒感染者體內(nèi)存在氧化應(yīng)激。慢性乙型肝炎患者外周血淋巴細(xì)胞內(nèi)IFNAR2的平均熒光強(qiáng)度與血漿GST水平的負(fù)相關(guān)趨勢提示氧化應(yīng)激對慢性乙型肝炎患者外周血IFNAR的表達(dá)起重要調(diào)節(jié)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.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R512.62

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本文編號：1540735