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目的探讨视黄酸(RA)对儿童淋巴结B细胞成熟分化的影响,及其与视黄酸受体基因表达水平变化的关系。方法本组24例,按年龄分为<1岁组、1～3岁组和～5岁组,每组8例。取患儿正常淋巴结分离细胞进行体外培养,分为RA组,RA+视黄酸受体α(RARα)拮抗剂(Ro415253)组(RA+Ro),脂多糖组(LPS),(LPS+RA)组和对照组。分别加入atRA、Ro415253、LPS。培养24、48h收集细胞。采用流式细胞术分析细胞表面标志,观察B细胞的成熟分化;RT荧光定量PCR测定视黄酸受体基因mRNA。结果<1岁组,RA组成熟B细胞的百分比明显高于对照组(24h:23%±5%vs17%±3%;48h:28%±6%vs22%±4%)(P均<0.05);LPS+RA组活化B细胞数明显高于LPS组(24h:82%±10%vs76%±8%;48h:83%±8%vs78%±10%)(P均<0.05)。同时,RARα基因表达水平也显著上调,而该受体拮抗剂可抑制RA的调节作用。1～3岁组淋巴结B细胞的成熟和活化呈现完全相同的调节变化,而～5岁组RA的这些调节作用不明显。结论RA能促进体外培养的淋巴结B细胞的分化发育和活化,该作用在3岁以下小儿明显。RARα基因的表达调节可能是介导RA作用的主要途径。
Objective To investigate the effect of retinoic acid (RA) on the maturation and differentiation of B lymphocytes in children and its relationship with the gene expression of retinoic acid receptor. Methods The group of 24 cases, according to age is divided into <1 year old group, 1 to 3 years old group and ~ 5 years old group, 8 cases in each group. The cells were divided into RA group, RA + Ro415253 group (RA + Ro), lipopolysaccharide group (LPS), (LPS + RA) Group and control group. Join atRA, Ro415253, LPS respectively. The cells were cultured for 24,48 h. Cell surface markers were analyzed by flow cytometry to observe the maturation and differentiation of B cells. RT-PCR was used to determine the mRNA of retinoic acid receptor. Results The percentage of mature B cells in RA group was significantly higher than that in control group (<23% ± 5% vs 17% ± 3%; 48 h: 28% ± 6% vs 22% ± 4% ). The number of activated B cells in LPS + RA group was significantly higher than that in LPS group (24h: 82% ± 10% vs76% ± 8%; 48h: 83% ± 8% vs78% ± 10%) (all P <0.05). At the same time, RARα gene expression level was also significantly increased, and the receptor antagonist can inhibit the regulation of RA. The maturation and activation of B lymphocytes from 1 to 3-year-old group showed exactly the same regulatory changes, but these regulatory effects were insignificant in ~ 5-year-old group. Conclusion RA can promote the differentiation, development and activation of lymph node B cells cultured in vitro, which is obvious in children under 3 years of age. RARα gene expression regulation may be the main way to mediate the role of RA.