摘要
目的本研究旨在探讨生命早期脂多糖(LPS)对脐血CD4+T细胞白介素-13(IL-13)和白介素-17(IL-17)表达的影响及1,25-(OH)2D3对其表达的干预作用,为维生素D的临床合理应用及其对哮喘等变态反应性疾病的防治提供理论依据。方法选取顺产的足月新生儿12例,断脐后立即取胎盘端脐静脉血50 mL,采用密度梯度离心法分离脐血单个核细胞(CBMCs),磁珠分选CD4+T细胞后依据不同的处理方法分为空白刺激组、LPS(10μg/mL)单独刺激组和LPS(10μg/mL)+1,25-(OH)2D3(10-8mmol/L)共刺激组。培养72 h后采用双抗夹心酶联免疫吸附试验(ELISA)和Real-Time PCR分别检测培养上清液中IL-13和IL-17水平及CD4+T细胞中IL-13和IL-17 mRNA表达。结果与空白刺激组相比,LPS单独刺激组培养上清液中IL-13和IL-17水平及CD4+T细胞中IL-13和IL-17 mRNA表达水平明显升高(均P<0.01),而1,25-(OH)2D3处理可降低其表达水平(均P<0.05),但仍高于空白刺激组(均P<0.01)。结论 LPS可促进脐血CD4+T细胞IL-13和IL-17的表达;1,25-(OH)2D3对于LPS诱导的脐血CD4+T细胞IL-13和IL-17表达具有抑制作用,提示1,25-(OH)2D3可能在变应原致敏的早期发挥一定保护作用。
Objective To study the effect of 1,25-（OH）2D3 on lipopolysaccharide （LPS）-induced expression of interleukin-13 （IL-13） and interleukin-17 （IL-17） in cord blood CD4＋T ceils, providing theoretical basis for clinical reasonable application of vitamin D and prevention of asthma and allergic diseases. Methods Mononuclear cells （MNCs） were isolated from umblilical cord blood （50 mL） of 12 normal eutocia term newborns by gravity centrifugation. CD4~T cells were isolated using magnetic beads, which was cultured with following three kinds of stimulus for 72 hours：, natural state （blank group）, LPS （10 btg/mL）stimulation alone and LPS（10 txg/mL）＋l,25-（OH）2D3 （10-s mmol/L） stimulation. Levels oflL-13 and IL-17 in the culture supematant and mRNA expressions in cord blood CD4＋T cells were detected using ELISA and real Time-PCR respectively. Results Compared with the blank group, levels of IL-13 and 1L-17 in the culture supematant and mRNA expression of IL-13 and IL-17 in the cord blood CD4＋T ceils increased in the LPS stimulation alone group （P〈O.O1）. When co-stimulation of 1,25-（OH）2D3 with LPS, levels of IL-13 and IL-17 in the culture supematant and mRNA expression of IL-13 and IL-17 in the cord blood CD4＋T cells decreased compared with LPS-stimulated alone group （P〈0.05）, but remained higher than the blank group （P〈0.01）. Conclusions LPS can promote expression of IL-13 and IL-17 in cord blood CD4＋T cells. 1,25-（OH）2D3 inhibits the expression of IL-13 and IL- 17, suggesting that vitamin D intake may provide protective effects in the development of atopy-predisposing immuneresponses in early life.
出处
《中国当代儿科杂志》
CAS
CSCD
北大核心
2013年第9期763-766,共4页
Chinese Journal of Contemporary Pediatrics
基金
上海市科学技术委员会重大项目(10D21951100)
