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目的探讨外周血辅助性T细胞17(Th17)细胞水平在儿童脓毒症发病中的作用及临床价值。方法选取脓毒症患儿36例;健康儿童20例为正常对照组。分离外周淋巴细胞后,分别采用流式细胞仪检测Th17细胞比例;实时荧光定量PCR(qRT-PCR)技术检测Th17细胞特异性转录因子即维甲酸受体相关孤儿受体γt(RORγt)mRNA表达水平;酶联免疫吸附法(ELISA)检测血清IL-6,IL-17及IL-23的含量。结果与健康对照组比较,脓毒症患儿组外周血Th17细胞比例增高(t=3.68,P<0.05);脓毒症患儿组RORγt mRNA表达水平也显著高于健康对照组(t=8.32,P<0.05);同时脓毒症患儿组细胞因子含量(IL-6,IL-17,IL-23)明显高于健康对照组(P<0.05)。此外,脓毒症死亡组患儿外周血Th17细胞水平(2.2±0.5)%显著高于存活组(0.8±0.3)%(t=4.67,P<0.05),外周血Th17细胞比例与患儿死亡率呈正相关(r=0.24,P<0.001)。结论 Th17细胞参与了脓毒症的炎症反应,检测Th17细胞水平有助于儿童脓毒症的早期诊断、治疗及预后的判断。
Objective To investigate the role and clinical value of peripheral blood T helper 17 (Th17) cells in the pathogenesis of sepsis in children. Methods Thirty-six children with sepsis were selected and 20 healthy children were selected as normal control group. After the peripheral lymphocytes were separated, the proportion of Th17 cells was detected by flow cytometry. The mRNA expression level of Th17 cell-specific orphan receptor γt (RORγt) mRNA was detected by real-time quantitative PCR (qRT-PCR) Serum levels of IL-6, IL-17 and IL-23 were detected by enzyme linked immunosorbent assay (ELISA). Results Compared with the healthy control group, the percentage of Th17 cells in sepsis group was significantly higher (t = 3.68, P <0.05). The expression of RORγt mRNA in sepsis group was significantly higher than that in healthy control group (t = 8.32 , P <0.05). At the same time, the levels of cytokines (IL-6, IL-17, IL-23) in children with sepsis were significantly higher than those in healthy controls (P <0.05). In addition, the level of Th17 cells (2.2 ± 0.5)% in the peripheral blood of the septic death group was significantly higher than that of the survival group (0.8 ± 0.3)% (t = 4.67, P <0.05) The rates were positively correlated (r = 0.24, P <0.001). Conclusion Th17 cells are involved in the inflammatory response of sepsis. Detecting the level of Th17 cells is helpful for the early diagnosis, treatment and prognosis of sepsis in children.