环孢霉素A对NIT-1胰岛β细胞基因表达谱的影响

Effects of cyclosporin A on gene expression profiles of NIT-1 pancreatic β cell line

目的采用基因芯片技术,观察免疫抑制剂环孢霉素A(CsA)对NIT-1胰岛β细胞基因表达谱的影响。方法体外培养NIT-1胰岛β细胞,以10μmol/LCsA处理24h。应用基因芯片分别检测CsA处理24h及空白对照组NIT-1胰岛β细胞的基因表达谱。结果CsA作用于NIT-1胰岛β细胞24h后,在4096条基因中,有38条基因表达上调,其中已知功能基因13条,主要是与应激反应、蛋白质合成及细胞生长相关的基因。有46条基因表达下调,其中已知功能基因25条,主要是与细胞生长发育、氧化磷酸化过程及蛋白合成有关的基因。RT-PCR技术验证了Zfr、Tpi和Pax63个基因表达的变化。结论CsA作用于NIT-1胰岛β细胞24h后,可影响NIT-1细胞中多种基因的表达,其中对细胞生长发育、氧化磷酸化等有关基因表达的抑制作用,可能与CsA抑制NIT-1细胞胰岛素释放的作用机制相关。本研究为进一步深入研究CsA对胰岛β细胞的作用机制提供了线索和依据。

Objective To observe the effects of cyclosporin A (CsA) on gene expression profiles of NIT-1 pancreatic β cells cell line using microarray technique.Methods NIT-1 cells were exposed to cyclosporin A treatment (10 μmol/L) for 24 h and the differential gene expressions were assessed using microarray technique.Results After 24h of CsA treatment,38 of the 4096 genes tested were up-regulated,including 13 genes with known functions involving stress response,cell growth and protein synthesis.Meanwhile 46 genes were down-regulated,including 25 genes with known functions involving cell growth and maturation,oxidative phosphorylation and protein synthesis.Changes of gene expression of Zfr,Tpi and Pax6 were confirmed by semi-quantitative reverse transcription polymerase chain reaction.Conclusions CsA treatment for 24 h induces changes in the gene expression profiles of NIT-1 pancreatic β cells.Down-regulation of the genes related to cell growth and maturation,oxidative phosphorylation and protein synthesis may partly explain the mechanisms that CsA inhibits the release of insulin from pancreatic β cells.