目的检测高迁移率族蛋白BI(HMGB1)基因多态性在中国北京地区人群的分布特点,并分析其多态性与重要炎症介质诱生的关系。方法收集200例健康献血员外周全血,根据文献通过基因测序对中国北京人群HMGB1启动子区及外显子、外显子和内含子交界区的突变位点进行检测,同时对外周全血体外内毒素(LPs)刺激培养,通过实时定量PCR及酶联免疫吸附试验方法分析,对HMGB1、白介素(IL)~10、肿瘤坏死因子(TNF)-α、IL-6等细胞因子的基因表达和蛋白质分泌水平进行检测,并与HMGB1多态性的相关关系进行分析。结果中国北京地区人群HMGB1存在四种多态性:即rs1412125、rs2249825、rs3742305、rs41376448。在LPS体外刺激后仅rs2249825多态性与HMGB1诱生水平相关,P〈0.05,但基因水平差异无统计学意义,P〉0.05;而其他三种多态性在HMGB1、IL-10、TNF-α、IL-6蛋白质及基因水平均无统计学的意义,均P〉0.05。结论中国北京人群HMGB1的基因有四种多态性,LPS刺激后HMGB1的诱生水平与rs2249825具有相关性,它可能影响HMGB1蛋白质水平的表达。
Objective To investigate the distribution characteristics of high mobility group box-1 protein (HMGB1) gene polymorphisms in the population of Beljing, and locate the relationship between the polymorphisms and formation of the inflammatory cytokines. Methods A total of 200 healthy individuals from the population in Beijing were enrolled in the study. Peripheral blood samples were collected to determine the mutations in HMGB1 promoter and exon, and the area between exon and intron in by the genomes sequencing according to relative literatures. Then, peripheral blood was stimulated by lipopolysaccharide (LPS) in vitro, gene expression and protein secretion levels of HMGBl, interleukin (IL)-10, tumor necrosis factor (TNF)-α, and IL-6 were measured by real-time PCR and ELISA. In addition, the relationship between gene as well as protein expression of those inflammatory eytokines and HMGB1 polymorphisms was analyzed. Results There were four polymorphisms in HMGB 1 genes in Beijing' s population: rs1412125, rs2249825, rs3742305, and rs41376448. Of them, only rs2249825 was correlated with protein levels of HMGB1 in blood samples stimulated with LPS in vitro, but not in the gene levels. As to the other three polymorphisms, no significant statistical differences were noticed after stimulation with LPS in both protein and gene levels of HMGB1, IL-10, TNF-α, and IL-6. Conclusion Among the four HMGB1 polymorphisms in the population in Beijing, only rs2249825 is associated with the production of HMGB1 protein level after LPS stimulation.