B、C基因型乙肝病毒在HEK-293T细胞中免疫炎症反应比较研究
发布时间:2018-05-05 01:11
本文选题:乙肝 + 基因型 ; 参考:《南昌大学》2017年硕士论文
【摘要】:目的:通过观察比较B、C基因型乙肝病毒感染HEK-293T细胞后分泌细胞因子产生的免疫炎症反应差异,作为B和C基因型乙肝病毒和临床病情是否相关的参考,从而对乙肝基因型与临床相关性的发生机制作补充,为临床治疗提供方向。方法:随机选取2015年1月至2015年9月至南昌市第九医院门诊和住院就诊的慢性乙型肝炎患者(HBV-DNA均为阳性)的血清标本254例。性别、年龄不限。并排除其他合并的疾病,且未进行性抗病毒治疗。诊断符合2015年中华医学会肝病学分会,中华医学会感染病学分会联合制订的慢性乙型肝炎防治指南。采用多对型特异性引物巢式PCR法鉴定并筛选出血清标本中的B、C两种基因型,随机选取B、C基因型15例。分别扩增B、C基因型血清标本中HBV病毒的s和c基因片段。采用克隆技术构建含有s和c基因片段表达绿色荧光蛋白的重组真核表达质粒,转染至HEK-293T细胞中,检测该细胞中IL-1β、TNF-α等的细胞因子分泌的水平。分析各组间的临床特征、IL-1β、TNF-α水平的差异。采用SPSS20.0软件进行统计学分析。P0.05具有统计学意义。结果:转染B基因型的s片段细胞中IL-1β水平为(143.89±46.48)pg/ml、TNF-α水平为(7.97±1.03)pg/ml,c片段细胞中IL-1β水平为(253.62±65.12)pg/ml、TFN-α水平为(8.65±1.07)pg/ml;C基因型的s片段中IL-1β水平为(40.96±29.29)pg/ml,TNF-α水平为(4.22±0.56)pg/ml,c片段中IL-1β水平为(35.78±19.31)pg/ml,TNF-α水平为(3.39±0.33)pg/ml。结论:乙肝病毒B基因型中基因片段在体外HEK-293T中的免疫炎症介质表达量明显高于C基因型。这或许是一个与临床致病性不同有关的因素。能从某种程度上证实两种基因型在体外细胞免疫反应过程中的不同。这为我们提供了另一个慢性乙型肝炎的研究角度。
[Abstract]:Objective: to observe the difference of cytokines produced by cytokines in HEK-293T cells infected with HBV genotype B and C, and to provide a reference for the relationship between hepatitis B virus genotype B and hepatitis B virus and clinical conditions. Thus, the mechanism of hepatitis B genotype and clinical correlation is supplemented, and the direction of clinical treatment is provided. Methods: 254 serum samples from outpatients and inpatients with chronic hepatitis B from January 2015 to September 2015 were randomly selected. Sex, age is unlimited. Other associated diseases were excluded without progressive antiviral therapy. The diagnosis is in accordance with the guidelines for the prevention and treatment of chronic hepatitis B developed jointly by the Society of Hepatology of the Chinese Medical Association and the Society of Infectious Diseases of the Chinese Medical Association in 2015. Two genotypes of BMC were identified and screened by nested PCR with multiple pairs of type-specific primers. 15 patients were randomly selected. The s and c gene fragments of HBV virus were amplified from the serum samples of genotype BU C, respectively. A recombinant eukaryotic expression plasmid containing s and c gene fragments expressing green fluorescent protein was constructed and transfected into HEK-293T cells to detect the level of cytokines secreted by IL-1 尾 -TNF- 伪. To analyze the clinical characteristics and the level of IL-1 尾 -TNF- 伪 in each group. The use of SPSS20.0 software for statistical analysis. P0.05 has statistical significance. 缁撴灉:杞煋B鍩哄洜鍨嬬殑s鐗囨缁嗚優涓璉L-1尾姘村钩涓,
本文编号:1845508
本文链接:https://www.wllwen.com/yixuelunwen/chuanranbingxuelunwen/1845508.html
最近更新
教材专著