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目的检测哮喘模型小鼠辅助性T17细胞(Th17)和调节性T细胞(Treg)相关细胞因子及受体的表达变化,探讨Th17/Treg在哮喘发病机制中的作用。方法 20只清洁级BALB/C雌鼠,鼠龄6周,体质量18~20 g。随机分为对照组和哮喘组,每组10只。哮喘组小鼠腹腔注射卵白蛋白(OVA)与氢氧化铝的氯化钠混悬液3次后,给予雾化的OVA激发,建立哮喘小鼠模型。对照组给予等量的0.9%氯化钠溶液。末次激发后24 h处死小鼠,检测支气管肺泡灌洗液(BALF)中细胞总数和细胞分类计数;酶联免疫吸附分析(ELISA)、实时定量聚合酶链反应(Q-PCR)和Western blot法测定小鼠外周血和肺组织中的Th17/Treg细胞相关因子及受体的表达水平。结果哮喘组小鼠BALF中细胞总数、嗜酸性细胞比例、中性粒细胞比例、淋巴细胞比例显著高于对照组。哮喘组外周血和肺组织中白细胞介素(IL)-17和IL-18高于对照组[IL-17(124.36±2.14)pg/mL、(21.70±1.02)pg/mL vs(25.98±5.23)pg/mL、(19.95±3.21)pg/mL;IL-18(227.06±24.14)pg/mL、(413.97±32.01)pg/mL vs(120.38±10.32)pg/mL、(238.21±15.33)pg/mL],IL-10和转化生长因子β(TGF-β)低于对照组[IL-10(60.42±24.12)pg/mL、(65.14±15.11)pg/mL vs(61.07±14.36)pg/mL、(104.36±28.16)pg/mL;TGF-β(38.72±12.12)pg/mL、(58.65±24.21)pg/mL vs(30.19±10.16)pg/mL、(121.06±39.12)pg/mL];哮喘组维甲酸相关孤儿素受体γt(RORγt)水平高于对照组[(8.77±2.35)、(4.46±1.05)vs(3.03±1.02)、(3.45±1.25)],肺叉头蛋白3(FoxP3)水平低于对照组[(5.15±1.84)vs(12.35±3.21)];并且外周血和肺组织中相关细胞因子及受体的表达差异具有显著统计学意义(P<0.01)。结论 Th17/Treg及其主要相关因子在哮喘的发病中具有重要作用。
Objective To detect the expressions of Th17 and Treg related cytokines and their receptors in asthmatic mice and to explore the role of Th17 / Treg in the pathogenesis of asthma. Methods Twenty female BALB / C mice of clean grade were aged 6 weeks and their body weight was 18-20 g. Randomly divided into control group and asthma group, 10 rats in each group. The asthmatic mice were intraperitoneally injected with ovalbumin (OVA) and sodium hydroxide suspension of aluminum hydroxide three times and then challenged with OVA to establish an asthmatic mouse model. The control group was given the same amount of 0.9% sodium chloride solution. Twenty-four hours after the final challenge, mice were sacrificed and the total number of cells and the number of cells in the bronchoalveolar lavage fluid (BALF) were determined. ELISA, Q-PCR and Western blot The levels of Th17 / Treg cell related factors and receptors in peripheral blood and lung tissue of mice were determined. Results The total number of cells, eosinophils, neutrophils and lymphocytes in BALF in asthma group were significantly higher than those in control group. The levels of IL-17 and IL-18 in peripheral blood and lung tissue of asthma group were significantly higher than that of the control group [IL-17 (124.36 ± 2.14) pg / mL, (21.70 ± 1.02) pg / mL vs (25.98 ± 5.23 ), pg / mL (19.95 ± 3.21) pg / mL, IL-18 (227.06 ± 24.14) pg / mL, (413.97 ± 32.01) pg / mL vs (120.38 ± 10.32) pg / / mL], IL-10 and TGF-β were lower than those of the control group [IL-10 (60.42 ± 24.12) pg / mL, (65.14 ± 15.11) pg / mL vs (61.07 ± 14.36) pg / mL, (104.36 ± 28.16) pg / mL, TGF-β (38.72 ± 12.12) pg / mL, (58.65 ± 24.21) pg / mL vs (30.19 ± 10.16) pg / mL, (8.77 ± 2.35), (4.46 ± 1.05) vs (3.03 ± 1.02), (3.45 ± 1.25), respectively, compared with the control group (P <0.05) (5.15 ± 1.84 vs 12.35 ± 3.21). The expression of related cytokines and their receptors in peripheral blood and lung tissue were significantly different (P <0.01). Conclusion Th17 / Treg and its major related factors play an important role in the pathogenesis of asthma.