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目的:检测中性粒细胞亚型哮喘(NA)患者痰上清中辅助性T细胞17(Th17)特异性转录因子维甲酸相关核孤儿受体γt(RORγt)、B细胞转录激活因子(BATF)及外周血中Th17细胞相关细胞因子白细胞介素8(IL-8)、白细胞介素17(IL-17)和白细胞介素22(IL-22)表达水平,探讨其在NA发病中的作用。方法:以56例支气管哮喘患者为研究对象,4.5%生理盐水雾化诱导痰,经细胞MGG染色进行哮喘的炎症亚型分型,分为嗜酸性粒细胞亚型哮喘(EA)组(26例)和NA组(30例),以常规体检者为健康对照组(NC组,28人)。实时荧光定量PCR(RT-PCR)法检测痰上清中BATF和RORγt mRNA表达水平。空腹抽取各组研究对象肘静脉血10mL,Ficoll密度梯度离心后分离外周血单个核细胞(PBMC),ELISA法测定血清中IL-8、IL-17和IL-22蛋白表达水平,流式细胞术检测PBMC中Th17细胞(CD4+IL-17+细胞)的百分率。结果:与NC组和EA组比较,NA组患者血清中IL-8、IL-17和IL-22表达水平均明显升高(P<0.05);与NC组比较,EA组患者血清中IL-8、IL-17和IL-22表达水平差异无统计学意义(P>0.05)。与NC组和EA组比较,NA组患者痰上清中BATF和RORγt mRNA表达水平明显升高(P<0.01)。与NC组和EA组比较,NA组患者PBMC中Th17细胞百分率均明显升高(P<0.01);与NC组比较,EA组患者PBMC中Th17细胞百分率差异无统计学意义(P>0.05)。结论:IL-17转录因子RORγt和BATF参与了NA患者气道炎症发生过程,且Th17细胞及其相关细胞因子IL-17和IL-22的异常表达与NA的全身反应有关联。
AIM: To detect the expression of RORγt, B cell transcriptional activator (BATF) mRNA and protein of 17 T helper 17 (Th17) specific transcription factor in sputum supernatant of neutrophil subtype asthma (NA) (IL-8), interleukin-17 (IL-17) and interleukin-22 (IL-22) in peripheral blood of Th17 cells and explore its role in the pathogenesis of NA. Methods: Fifty-six patients with bronchial asthma were enrolled in this study. Phosphates were induced by 4.5% saline and sputum was induced by MGG staining. The subtypes of asthma were divided into eosinophil subtype asthma group (n = 26) ) And NA group (30 cases), and the control group was normal control group (NC group, 28). The expression of BATF and RORγt mRNA in sputum supernatant was detected by real-time fluorescence quantitative PCR (RT-PCR). Fasting blood samples were collected from 10 mL of elbow venous blood in each group. Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll density gradient centrifugation. Serum levels of IL-8, IL-17 and IL-22 were measured by ELISA. The percentage of Th17 cells (CD4 + IL-17 + cells) in PBMCs was measured. Results: Compared with NC group and EA group, the levels of IL-8, IL-17 and IL-22 in NA group were significantly increased (P <0.05). Compared with NC group, the levels of IL- 8, IL-17 and IL-22 expression difference was not statistically significant (P> 0.05). Compared with NC group and EA group, BATF and RORγt mRNA levels in sputum supernatant of NA group were significantly increased (P <0.01). Compared with NC group and EA group, the percentage of Th17 cells in PBMC of NA group was significantly increased (P <0.01). Compared with NC group, the percentage of Th17 cells in PBMC of EA group was not significantly different (P> 0.05). CONCLUSIONS: IL-17 transcription factors RORγt and BATF are involved in the pathogenesis of airway inflammation in NA patients. The abnormal expression of Th17 cells and related cytokines IL-17 and IL-22 is associated with the systemic reaction of NA.