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目的:检测中国人群Toll样受体4(Toll-like receptor4,TLR4)基因5’调控区的单核苷酸多态性(single nucleotide polymorphisms,SNPs),探讨其与TLR4蛋白表达的关系。方法:采用聚合酶链反应-限制性片段长度多态性法(PCR-RFLP)对正常汉族人群样本TLR4启动子区-2242、-1892和-1837这3个可能有意义的SNP位点进行基因分型,以确定中国人群中TLR4基因启动子区SNP基因型和发生频率。取其中89例全血标本用全血培养模型检测内毒素刺激前后TLR4蛋白的表达变化,进一步探讨TLR4启动子区单核苷酸多态性对其蛋白表达的影响。结果:TLR4启动子区-2242、-1892和-1837这3个可能有意义的SNP位点等位基因频率分别是43.27%、27.70%和42.75%。TLR4蛋白表达检测结果表明内毒素刺激后-2242位点TC与CC基因型TLR4蛋白的表达显著高于TT基因型(P<0.05),其它位点则没有影响。结论:中国汉族人群中TLR4基因启动子区-2242位点可能是脓毒症关联分析重要的遗传标记。
OBJECTIVE: To detect single nucleotide polymorphisms (SNPs) in the 5 ’regulatory region of Toll-like receptor 4 (TLR4) gene in Chinese population and to explore its relationship with TLR4 protein expression. Methods: PCR-RFLP was used to detect the possible SNP loci of -2242, -1892 and -1837 in TLR4 promoter region of normal Han population Typing to determine the SNP genotype and frequency of TLR4 promoter in Chinese population. Totally 89 samples of whole blood were used to detect the expression of TLR4 protein before and after endotoxin stimulation in whole blood culture model to further explore the effect of single nucleotide polymorphism of TLR4 promoter on the protein expression. Results: The allele frequencies of three possible SNP SNPs at -2242, -1892 and -1837 in TLR4 promoter region were 43.27%, 27.70% and 42.75%, respectively. The results of TLR4 protein expression showed that the expression of TLR4 protein in TC genotype CC and CC genotype at -2242 was significantly higher than that in TT genotype (P <0.05) after stimulation with endotoxin, but not in other sites. CONCLUSION: The -2242 locus of TLR4 gene promoter in Chinese Han population may be an important genetic marker for sepsis association analysis.