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目的探讨转化生长因子β1(TGFβ1)基因多态性与乙型肝炎肝硬化的关系及其对血浆TGFβ1浓度的影响。方法分别应用聚合酶链反应扩增难控性突变系统(PCR ARMS)和Lightcycler结合测序的方法,测定了92名健康对照者和134例肝硬化患者TGFβ1基因-988、-800、-509、codon10、codon25和codon263位点的单核苷酸多态性,并确定了其基因型和等位基因频率的分布;用ARLEQUNVer2.0软件分析了-509位点和codon10位点等位基因之间是否存在连锁不平衡;用ELISA的方法检测了血浆中TGFβ1和Ⅳ型胶原的浓度,用放射免疫法检测了血浆中透明质酸和Ⅲ型前胶原N端肽。结果本研究中TGFβ1基因-988、-800、codon25和codon263位点不存在基因多态性;-509位点基因型及等位基因分布频率在肝硬化组和正常对照组中差异无统计学意义,codon10TT基因型及等位基因T在肝硬化组中的分布频率(0.313,0.537)明显高于对照组(0.196,0.440)。把肝硬化患者按Child Pugh分级分为A、B、C3组,-509CC基因型在肝硬化C级组中的分布频率(0.631)明显高于TT基因型(0.325),codon10位点基因多态性在肝硬化的各个分级组中的分布频率差异无统计学意义。肝硬化组血浆TGFβ1、Ⅳ型胶原、透明质酸及Ⅲ型前胶原N端肽的浓度显著高于对照组。在对照组中,血浆TGFβ1浓度在-509位
Objective To investigate the relationship between transforming growth factor β1 (TGFβ1) gene polymorphism and hepatitis B cirrhosis and its effect on plasma TGFβ1 concentration. Methods The mRNA and protein expressions of TGFβ1, -988, -800, -509 and codon10 in 92 healthy controls and 134 cirrhotic patients were determined by PCR ARMS and Lightcycler sequencing. , Codon25 and codon263 sites and determined their genotypes and allele frequency distributions. The ARLEQUNVer2.0 software was used to analyze whether the -509 and codon10 alleles were There was linkage disequilibrium. The concentration of TGFβ1 and type Ⅳ collagen in plasma was detected by ELISA, and the plasma hyaluronic acid and type Ⅲ procollagen N-terminal peptide were detected by radioimmunoassay. Results There was no gene polymorphism at sites of -988, -800, codon25 and codon263 in TGFβ1 gene in this study. There was no significant difference in genotype and allele frequencies at -509 between cirrhosis and normal controls , codon10TT genotype and allele T frequency distribution in the cirrhosis group (0.313,0.537) was significantly higher than the control group (0.196,0.440). According to Child Pugh classification, patients with liver cirrhosis were divided into A, B and C3 groups. The distribution frequency of -509CC genotype in cirrhosis C group (0.631) was significantly higher than TT genotype (0.325), codon10 polymorphism There was no significant difference in the distribution frequency of sex in each grading group of cirrhosis. The levels of plasma TGFβ1, type Ⅳ collagen, hyaluronic acid and type Ⅲ procollagen N-terminal peptide in cirrhosis group were significantly higher than those in control group. In the control group, the plasma TGFβ1 concentration was at -509