摘要
目的:研究视黄酸调节胸腺细胞发育的作用及其机制。方法:采用体外胸腺组织培养体系,在培养体系中加入视黄酸或视黄酸受体拮抗剂。在培养的不同时间收集胸腺细胞,用荧光标记的抗CD3、CD4、CD8抗体染色,并进行流式细胞仪分析,观察胸腺细胞表面标志随成熟分化的变化。胸腺细胞抽提的RNA采用实时定量PCR法检测视黄酸受体α(RARα) mRNA的表达。结果:体外胸腺组织培养体系能支持胸腺细胞的发育成熟。在培养24 h促进未成熟的CD4+CD8+胸腺细胞向CD4+成熟细胞分化发育作用明显,并且该作用可被RARα特异性拮抗剂Ro41-5253所拮抗。视黄酸也能显著抑制CD4+CD8+胸腺细胞向CD8+细胞分化,该作用也在培养24 h出现。实时定量PCR显示,视黄酸能促进RARα mRNA表达,并且 RARαmRNA表达水平与CD4+CD8+胸腺细胞百分数之间存在正相关,与CD8+胸腺细胞百分数之间存在负相关。结论:视黄酸可能通过影响其受体RARα表达而影响胸腺细胞的成熟分化。
Objective :To study the effect and mechanism of retinoic acid(RA) on thymocytes development. Methods:Thymus tissue culture system was used to investigate the roles of RA in maturation development of thymocytes. Thymus tissues were cultured with or without retinoic acid and/or retinoic acid receptor antagonist. The cultures were harvested after 24, 48 and 72 hours, respectively. Thymocytes were stained with fluorescence labeled anti-CD3, anti-CD4, anti-CD8 antibodies and analyzed by flow cytometry. The real time quantitative RT-PCR assay was used to detect the retinoic acid receptor (RAR) mRNA expression in thymocytes. Results:The thymus tissue culture system was able to support the maturation of thymocytes. Addition of RA to the culture system promoted thymocytes differentiation from immature CD4+CD8+ cells to mature CD4+ cells, but inhibited the transformation from immature CD4+CD8+ cells to mature CD8+ cells. The effect was marked after 24h culture, and was antagonized by Ro41-5253. RA enhanced the expression of RAR mRNA in thymocytes. A positive correlation was found between the expression of RAR mRNA and the percentage of CD4+CD8+ cells and a negative correlation between the expression of RAR mRNA and the percentage of CD8+ cells. Conclusion:RA likely play an important role in thymocytes development, perhaps by affecting the relative expression of RAR gene.
出处
《营养学报》
CAS
CSCD
北大核心
2004年第1期9-12,共4页
Acta Nutrimenta Sinica
基金
国家自然科学基金资助项目(No.30070661)
