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目的观察不同浓度地塞米松(DEX)和甲泼尼龙(MP)对支气管哮喘患儿外周血单个核细胞(PBMC)中CD4+T淋巴细胞中2个功能性亚群(Th1/Th2)功能状态的影响。方法选择2001年6月-2002年6月在重庆医科大学附属儿童医院就诊的15例哮喘患儿和14例健康体检儿童为研究对象。分为哮喘对照组、10-7mol.L-1DEX干预组、10-8mol.L-1DEX干预组、10-9mol.L-1DEX干预组、10-7mol.L-1MP干预组、10-8mol.L-1MP干预组、10-9mol.L-1MP干预组。清晨取其空腹静脉血5 mL,肝素抗凝,采用密度梯度离心法分离PBMC,加植物血凝素进行刺激培养,分别用10-7mol.L-1、10-8mol.L-1及10-9mol.L-1DEX或MP体外干预培养48 h。同期采集健康儿童空腹静脉血,同法分离培养。用ELISA法测定培养上清中γ干扰素(IFN-γ)、IL-4、IL-10及IL-12水平,并计算不同浓度DEX或MP对PBMC分泌细胞因子的抑制率。结果 1.哮喘对照组PBMC分泌IL-4水平显著高于健康对照组(P<0.05),IFN-γ/IL-4比值较健康对照组显著降低(P<0.05),2组间IFN-γ、IL-10及IL-12水平比较差异均无统计学意义(Pa>0.05)。2.DEX和MP均可明显抑制哮喘患儿PBMC分泌IFN-γ、IL-4及IL-12,但对IL-10抑制作用差;与健康对照组比较,哮喘组DEX和MP抑制PBMC分泌IL-10的作用弱,差异有统计学意义(Pa<0.05)。3.DEX和MP均以浓度依赖方式抑制哮喘患儿PBMC分泌IL-4,DEX在10-9mol.L-1时有促进IL-4分泌的效应,MP在10-9mol.L-1时可抑制IL-4的分泌,二组比较差异有统计学意义(P<0.05)。若以IFN-γ/IL-4表示Th1/Th2间的平衡,则MP可恢复Th1/Th2平衡(P<0.05)。结论 DEX和MP均可抑制Th1/Th2类细胞因子分泌,但MP有助于恢复Th1/Th2平衡,提示临床选择MP治疗支气管哮喘更有利。
Objective To observe the effects of different concentrations of dexamethasone (DEX) and methylprednisolone (MP) on the functional status of two functional subsets (Th1 / Th2) in peripheral blood mononuclear cells (PBMC) from children with bronchial asthma Impact. Methods From June 2001 to June 2002, 15 cases of asthmatic children and 14 healthy children were investigated in Children’s Hospital Affiliated to Chongqing Medical University. Divided into asthma control group, 10-7mol.L-1DEX intervention group, 10-8mol.L-1DEX intervention group, 10-9mol.L-1DEX intervention group, 10-7mol.L-1MP intervention group, 10-8mol. L-1MP intervention group, 10-9mol.L-1MP intervention group. In the early morning, 5 mL of fasting venous blood and heparin anticoagulation were taken. PBMCs were isolated by density gradient centrifugation and incubated with phytohaemagglutinin for 10-7mol.L-1, 10-8mol.L-1 and 10- 9mol.L-1DEX or MP in vitro intervention for 48 h. Fasting blood samples collected in healthy children at the same time, the same method of separation and culture. The levels of IFN-γ, IL-4, IL-10 and IL-12 in culture supernatants were determined by ELISA. The inhibitory rates of cytokines secreted by PBMC were calculated by different concentrations of DEX or MP. The level of IL-4 secreted by PBMC of asthma control group was significantly higher than that of healthy control group (P <0.05), the ratio of IFN-γ / IL-4 was significantly lower than that of healthy control group (P <0.05) , IL-10 and IL-12 levels were no significant difference (Pa> 0.05). Both DX and MP inhibited the secretion of IFN-γ, IL-4 and IL-12 from PBMC in asthmatic children, but the inhibitory effect on IL-10 was poor. Compared with the healthy control group, -10 weak role, the difference was statistically significant (Pa <0.05). Both DX and MP inhibited the secretion of IL-4 by PBMCs from children with asthma in a concentration-dependent manner. When DEX was 10-9mol.L-1, IL-4 secretion was promoted. When MP was at 10-9mol.L-1 Inhibition of IL-4 secretion, the difference between the two groups was statistically significant (P <0.05). When the balance between Th1 / Th2 was expressed as IFN-γ / IL-4, MP restored the Th1 / Th2 balance (P <0.05). Conclusions Both DEX and MP can inhibit the secretion of Th1 / Th2 cytokines, but MP can help restore the balance of Th1 / Th2, suggesting that clinical treatment of bronchial asthma with MP is more beneficial.