摘要
目的观察脓毒症患者静脉血Toll样受体4(TLR4)是否存在基因多态性,同时了解TLR4 mRNA、干扰素γ(IFN-γ)、白细胞介素23(IL-23)、降钙素原(PCT)、C反应蛋白(CRP)在不同严重程度脓毒症患者中的表达及对诊断的价值。方法收集受试者外周血提取基因组DNA,应用基因测序及酶切方法检测TLR4 Asp299Gly位点多态性,RT-PCR方法了解TLR4 mRNA表达,酶联免疫吸附测定法监测血清中IFN-γ、IL-23、PCT、CRP表达水平。结果(1)在TLR4基因Asp299Gly位点不存在基因多态性。(2)外周血TLR4 mRNA表达:正常组第1天与急性生理和慢性健康评分Ⅱ(APACHEⅡ)≤20组第1天、正常组第1天与APACHEⅡ>20组第1天、APACHEⅡ≤20组第1天与APACHEⅡ>20组第1天差异均有统计学意义(t=5.741、14.780、10.500,P值均<0.01)。正常组第7天与APACHEⅡ≤20组第7天、正常组第7天与APACHEⅡ>20组第7天、APACHEⅡ≤20组第7天与APACHEⅡ>20组第7天差异均有统计学意义(t=4.186、13.830、9.645,P值均<0.01)。(3)脓毒症患者第1天、第7天血IFN-γ、IL-23、PCT、CRP明显升高(P值均<0.01),TLR4与IFN-γ、IL-23呈正相关(P值均<0.01)。(4)TLR4、INF-γ及IL-23分别取891.6、84.5及861 μg/L为截断点,对发现脓毒症的敏感度均为100%,诊断的特异度为57%、57%及97%。结论(1)在TLR4基因Asp299Gly位点未发现多态性。(2)脓毒症患者TLR4 mRNA、IFN-γ、IL-23、PCT、CRP高表达,且随严重程度增加而增高。(3)TLR4 mRNA与IFN-γ、IL-23的表达可成为脓毒症的分子标记物。
Objective To observe the presence of toll-like receptor 4 (TLR4) gene polymorphism on the venous blood of patients with sepsis.Meanwhile to observe the expression and the value of the diagnosis of TLR4 mRNA, interferon-γ(IFN-γ), interleukin-23 (IL-23), procalcitonin (PCT) and C-reactive protein (CRP) on patients with different severity of sepsis.Methods The subjects in the peripheral blood were collected to extract genomic DNA, gene sequencing and enzyme digestion method is applied to detection of TLR4 Asp299Gly loci polymorphism.The mRNA expression of TLR4 mRNA in peripheral blood was detected by reverse transcriptase polymerase chain reaction (RT-PCR). The expression of IFN-γ, IL-23, PCT and CRP were detected by enzyme linked immunosorbent assay (ELISA).Results (1)The Asp299Gly site in sepsis do not exist polymorphism.(2)The expression of TLR4 mRNA in peripheral blood in patients with sepsis compared among different groups had significant differences on d1 [with Acute Physiology and Chronic Health Evaluation Ⅱ (APACHEⅡ)≤ 20 group and APACHEⅡ>20 group vs normal group, and APACHEⅡ≤20 group vs APACHEⅡ>20 group, t=5.741, 14.780 and 10.500, all P<0.01] and d7 (with APACHEⅡ≤20 group and APACHEⅡ>20 group vs normal group, and APACHEⅡ≤20 group vs APACHEⅡ>20 group, t=4.186, 13.830 and 9.645, all P<0.01). (3)Blood IFN-γ, IL-23, PCT and CRP of d1, d7 in patients significantly elevated (P<0.01). The concentration of IFN-γ, IL-23 were positively corelated with TLR4 mRNA concentration in sepsis.(4)The best cutoff value of TLR4 mRNA at baseline was 891.6 μg/L, with a sensitivity of 100%, specificity of 57%.The best cutoff value of IFN-γ at baseline was 84.5 μg/L, with a sensitivity of 100%, specificity of 57%.The best cutoff value of IL-23 at baseline was 861 μg/L, with a sensitivity of 100%, specificity of 97%.Conclusions (1)The Asp299Gly site in sepsis do not exist polymorphism.(2)The diagnostic rate of TLR4 mRNA, IFN-γ and IL-23 are relatively high in the diagnosis of sepsis.Patients with
作者
田铁英
徐喜媛
杨敬平
Tian Tieying;Xu Xiyuan;Yang Jingping(Department of Respiratory and Critical Care Medicine,Inner Mongolia Baogang Hospital, Baotou 014010,China)
出处
《国际呼吸杂志》
2019年第2期107-112,共6页
International Journal of Respiration
