脓毒症患者外周血单核细胞NLRP3炎性小体的表达研究

The study on NLRP3 inflammatory corpuscle expression in peripheral blood mononuclear cells from septic patients

目的:分析脓毒症患者外周血单核细胞(PBMCs)中核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)mRNA、半胱氨酸天冬氨酸酶-1(caspase-1)mRNA表达及下游炎症因子水平变化,探讨其在脓毒症发生发展中作用。方法:选取192例脓毒症患者为脓毒症组、187例非感染性全身性炎症反应综合征(SIRS)患者为SIRS组、184例来医院体检的健康人群为参照组。采用实时荧光定量聚合酶链反应(qPCR)检测NLRP3 mRNA、caspase-1 mRNA表达,酶联免疫吸附法(ELISA)测定血清降钙素原(PCT)、C反应蛋白(CRP)水平及血浆白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)水平。结果:脓毒症组患者血清PCT水平、血清CRP水平、APACHEⅡ及SOFA评分最高、SIRS组患者次之、参照组最低(P<0.05)。脓毒症患者NLRP3 mRNA、caspase-1 mRNA表达和血浆IL-1β、IL-18水平最高、SIRS组患者次之、参照组最低(P<0.05)。脓毒症患者NLRP3 mRNA表达与caspase-1 mRNA表达(P<0.05)、血浆IL-1β水平(P<0.05)、血浆IL-18水平(P<0.05)均呈正相关;且脓毒症患者NLRP3 mRNA、caspase-1 mRNA表达均与血清PCT水平、血清CRP水平、APACHEⅡ评分、SOFA评分呈正相关(P<0.05)。影响脓毒症患者预后的独立危险因素有血清PCT、CRP水平、APACHEⅡ评分、SOFA评分、NLRP3 mRNA表达、caspase-1 mRNA表达及血浆IL-1β、IL-18水平(P<0.05)。结论:脓毒症患者外周血单核细胞NLRP3 mRNA、caspase-1 mRNA高表达,血浆IL-1β、IL-18水平升高,NLRP3炎性小体参与脓毒症发生发展。

Objective:To analyze the changes of expressions of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)mRNA,cysteine-requiring aspartate protease-1(caspase-1)mRNA and downstream inflammatory factors levels in peripheral blood mononuclear cells(PBMCs)of patients with sepsis,and to explore their roles in the occurrence and development of sepsis.Method:One hundred and ninty-two patients with sepsis were selected as sepsis group,187 patients with non-infectious systemic inflammatory response syndrome(SIRS)as SIRS group,184 healthy people who came to the hospital for physical examination as control group.Real time fluorescence quantitative polymerase chain reaction(qPCR)was used to detect the expressions of NLRP3 mRNA and caspase-1 mRNA,the levels of serum procalcitonin(PCT),C-reactive protein(CRP),interleukin-1β(IL-1β)and interleukin-18(IL-18)were determined by enzyme-linked immunosorbent assay(ELISA).Result:The PCT level,CRP level,APACHE II score and SOFA score were the highest in sepsis group,followed by SIRS group and the lowest in control group(P<0.05).The expressions of NLRP3 mRNA and caspase-1 mRNA,levels of plasma IL-1βand IL-18 in sepsis patients were the highest,followed by SIRS patients and the lowest in control group(P<0.05).The expression of NLRP3 mRNA was positively correlated with caspase-1 mRNA(P<0.05),plasma IL-1β(P<0.05)and plasma IL-18(P<0.05);the expressions of NLRP3 mRNA and caspase-1 mRNA were positively correlated with serum PCT level,serum CRP level,APACHE II score and SOFA score(P<0.05).The independent risk factors influencing the prognosis of sepsis patients were serum PCT,CRP levels,APACHE II score,SOFA score,NLRP3 mRNA expression,caspase-1 mRNA expression,and plasma IL-1βand IL-18 levels(P<0.05).Conclusion:The expressions of NLRP3 mRNA and caspase-1 mRNA in peripheral blood mononuclear cells are high in sepsis patients,plasma IL-1 and IL-18 levels are increased,NLRP3 inflammatory corpuscles participate in the occurrence and development of sepsis.