摘要目的 观察CD4+ CD25+调节T细胞(Treg)/辅助性T细胞17(Th17)细胞在脓毒症大鼠炎性免疫反应中的作用.方法 110只雄性SD大鼠随机分为正常对照组、假手术组、脓毒症(CLP)组,采用改良的盲肠结扎穿孔术(CLP)制作大鼠脓毒症模型.采用流式细胞术检测CD14+单核细胞表面人类白细胞抗原-DR基因(HLA-DR)表达率、Treg细胞及TH17细胞比例;酶联免疫吸附试验(ELISA)检测白细胞介素(IL)-6、IL-10、肿瘤坏死因子(TNF)-α、转化生长因子(TGF)-β、白细胞介素(IL)-17炎性因子蛋白表达.结果 与假手术组比较:(1)伴随着脓毒症病情的发展,大鼠出现明显的免疫抑制,CD14+单核细胞HLA-DR表达率＜30％,IL-10/TNF-α比值(27.41 ±7.04比6.63 ±2.60)明显增高(P＜0.01).(2)术后96 h脓毒症大鼠Treg细胞[(11.91±3.88)％比(6.57±2.60)％,P＜0.01]和Th17细胞[(5.14±0.29)％比(2.85±0.07)％,P＜0.01]表达明显增高.(3)术后96 h脓毒症组前炎性细胞因子IL-6[(42.31±15.89) ng/L比(6.32 ±3.18) ng/L,P＜0.01]、IL-10[(69.89 ±20.78) ng/L比(13.58±5.37) ng/L,P＜0.01]、TNF-α[(5.03±3.10) ng/L比(2.77±1.10) ng/L,P＜0.01]、TGF-β[(4.99±2.01) ng/L比(1.88±1.07) ng/L,P＜0.01]、IL-17[(92.77±11.64) ng/L比(7.58±2.30) ng/L,P＜0.01]表达明显增高.结论 伴随着脓毒症病情的发展,大鼠出现明显的免疫抑制;在大鼠脓毒症的发生发展中,Treg细胞介导的免疫抑制及Th17细胞介导免疫激活反应同时存在;脓毒症细胞因子微环境变化可能是导致Treg细胞/Th17细胞失衡的原因之一.

abstractsObjective To investigate the roles of CD4+ CD25 + regulatory T cells (Treg) and T helper type 17 (Th17) cells in inflammatory response in septic rats.Methods A total of 110 healthy male Sprague-Dawley rats (250-300 g) were randomly divided into 3 groups:normal control group,sham group and cecal ligation and puncture (CLP) group.The rat models of sepsis were established by improved CLP.Flow cytometric analysis was performed to detect the percentage of CD14 + T cell,Th17 cells and Treg cells subpopulation.Expressions of proinflammatory cytokines [interleukin (IL)-6,IL-10,tumor necrosis factor (TNF)-α,transforming growth factor (TGF)-β,IL-17] were measured by using enzyme-linked immunosorbent serologic assay.Results Compared with sham group,(1) While the spesis becoming worse,the rats were immunological inhibition,the expression levels of leucocyte antigen-DR (19.63 ± 7.56) ％ lower than 30％ and the rate of IL-10/TNF-α (27.41 ± 7.04 vs.6.63 ± 2.60,P ＜0.01) significantly up-regulated.(2) After 96 hours,the proportions of Treg cells [(11.91 ± 3.88)％ vs.(6.57±2.60)％,P＜0.01] andTh17 cells [(5.14 ±0.29)％ vs.(2.85 ±0.07)％,P ＜0.01] in sepsis group were significantly higher.(3) After 96 hours,the expression levels of IL-6 [(42.31 ±15.89) ng/Lvs.(6.32 ±3.18) ng/L,P＜0.01],IL-10 [(69.89 ±20.78) ng/L vs.(13.58±5.37) ng/L,P＜0.01],TNF-α [(5.03 ±3.10) ng/L vs.(2.77 ±1.10) ng/L,P＜ 0.01],TGF-β [(4.99±2.01) ng/L vs.(1.88±1.07) ng/L,P＜0.01],IL-17 [(92.77±11.64) ng/L vs.(7.58 ±2.30) ng/L,P ＜0.01] were significantly up-regulated in sepsis group.Conclusion While the spesis becoming worse,the rats were immunological inhibition.Aberrant activation of Th17 cells and Treg cells might be involved in pathogenesis in sepsis.The changes of cytokine environment in sepsis may be one of the factors causing the imbalance of Treg cells/Th17 cells.