微小RNA调控脓毒症患者外周血免疫因子表达及其机制研究

目的:探讨脓毒血症患者血清外周白细胞中的微小RNA(mi RNA)表达水平的变化及其在脓毒症患者中的表达意义以及免疫调控的关系。方法:采用流式细胞仪检测外周血CD4+CD25+Treg细胞表达,采用实时定量PCR(RT-PCR)方法检测110例脓毒症患者以及100例正常对照外周血白细胞中mi RNA以及Foxp3 m RNA表达量,酶联免疫吸附法测定TNF-α和IL-10浓度,序贯器官衰竭估计(SOFA)评分系统评价脓毒症患者的严重程度。对mi RNA与白细胞总数、TNF-α、IL-10和SOFA评分之间的相关性进行分析。结果:实验组mi RNA表达水平较对照组显著降低(P0.01),WBC、IL-10水平显著升高(P0.01)。实验组mi RNA表达水平以及SOFA评分、血清TNF-α和IL-10之间呈负相关关系(r值分别为-0.512、-0.623、-0.432,P0.05);与WBC无显著相关性(r=0.215,P0.05)。脓毒症患者外周血Treg表达率和Foxp3m RNA均显著高于对照组(P0.01);随病情严重而升高,轻、中、重度实验组间两两比较差异均有统计学意义(P0.01)。死亡均在重度脓毒症患者中,死亡组Treg、Foxp3 m RNA及IL-10均显著高于存活组(P0.01);mi RNA低于存活组(P0.01)。结论:脓毒症患者外周血的mi RNA表达量显著降低,表达水平在一定程度上可以反应机体的炎症反应情况,同时还可以判断疾病的严重度,且mi RNA参与对Treg细胞增殖的调节,在脓毒症免疫失衡机制中发挥一定的作用。

Clinical Significance of Small RNA Expression in Peripheral Blood of Patients with Sepsis and Its Influence on Immune Regulation Mechanism

Objective:To observe the clinical significance of Small RNA expression in peripheral blood of patients with sepsis and its influence on immune regulation mechanism.Methods:110 cases of sepsis patients were selected as study group and another 100 healthy cases were as controls. Flow cytometry instrument was used to detect peripheral blood CD4 + CD25 + Treg cells expression, real-time quantitative PCR (RT-PCR) was used to detect miRNA and Foxp3 mRNA expression in the peripheral blood, enzyme-linked immunosorbent was to detect concentration of TNF-alpha and IL-10, and sequential organ failure estimation (SOFA) score system was used to evaluate the severity of the patients with sepsis. The correlation of miRNA with total number of white blood cells, the TNF-alpha, IL-10 and SOFA score were analyzed.Results:The expression level of miRNA in study group was lower than that of the control group (P<0. 01), and the level of WBC and IL-10 was significantly increased (P<0.01). miRNA expression and SOFA score, serum TNF-alpha in patients with sepsis had a negative correlation with IL-10 (r = -0.512 -0.623, -0.432, P<0.05); there was no significant correlation with the WBC (r=0.215, P>0.05). Treg expression rate and Foxp3mRNA in peripheral blood of patients with sepsis were significantly higher than that of healthy controls (P<0.01) and increased along with the severity of disease, and there was statistical significance in comparison between slight, moderate and severe sepsis (P<0.01). Death patients appeared in the patients with severe sepsis, Treg, Foxp3 mRNA and IL-10 levels in death patients were significantly higher than survival patients (P<0.01); miRNA in death patients was lower than that of survival patients (P<0.01).Conclusion:MiRNA expression is decreased significantly in peripheral blood of patients with sepsis, miRNA level can not only reflect the state of inflammatory response of the body, and to a certain extent, can be used as index to judge the severity of disease and prognosis, miRNA is also involved in regulation of Treg cells proliferation, and play a role in sepsis immune mechanism.