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目的探讨CD14基因启动子-159C/T基因多态性对全血培养CD14mRNA表达及可溶性CD14(sCD14)浓度的影响。方法采集118例健康献血员血标本,用全血细胞培养模型检测内毒素刺激前后CD14mRNA表达及sCD14浓度的变化。采用聚合酶链反应(PCR)及限制性内切酶HaeⅢ对PCR产物的消化作用检测CD14基因多态性。同时,对内毒素刺激后肿瘤坏死因子α(TNF-α)诱生水平进行了分析。结果118例健康献血员中,等位基因T和C的频率分别为60.2%和39.8%;40例是T等位基因纯合子(TT),62例为杂合子(TC),还有16例基因型为CC。基因型TT与TC白细胞中CD14mRNA的表达及上清液sCD14浓度均明显高于CC纯合子(P<0.05或0.01)。并且TT纯合子TNF-α诱生水平为(352±215)pg/ml,显著高于基因型TC及CC[(261±163)pg/ml及(198±122)pg/ml,P<0.05]。结论内毒素受体CD14-159C/T基因多态性对全血培养CD14的表达及释放产生显著影响,并与内毒素诱导TNF-α的反应性相关。
Objective To investigate the effect of CD14 gene promoter-159C / T polymorphism on the expression of CD14 mRNA and the concentration of soluble CD14 (sCD14) in whole blood. Methods A total of 118 blood samples were collected from healthy donors. The changes of CD14 mRNA expression and sCD14 concentration before and after endotoxin stimulation were detected by whole blood cell culture. CD14 gene polymorphism was detected by polymerase chain reaction (PCR) and restriction endonuclease Hae Ⅲ digestion of PCR products. At the same time, the levels of tumor necrosis factor alpha (TNF-α) induced by endotoxin were analyzed. Results Among 118 healthy blood donors, the frequencies of alleles T and C were 60.2% and 39.8% respectively; 40 were TT homozygotes (T), 62 were heterozygous (TC) and 16 The genotype is CC. CD14 mRNA expression in TT and TC leukocytes and the concentration of sCD14 in supernatant were significantly higher than those in CC homozygotes (P <0.05 or 0.01). (352 ± 215) pg / ml, which was significantly higher than that of genotype TC and CC [(261 ± 163) pg / ml and (198 ± 122) pg / ml) ]. Conclusion The polymorphism of endotoxin receptor CD14-159C / T has a significant effect on the expression and release of CD14 in whole blood and is related to the reactivity of TNF-α induced by endotoxin.