摘要
目的研究免疫抑制剂环孢霉素A(CsA)对NIT-1胰岛β细胞胰岛素分泌的影响及在基因水平上对线粒体氧化磷酸化水平的影响。方法体外培养NIT-1胰岛β细胞,用10 μmol/L CsA处理24和48 h。放射免疫测定法(RIA)检测胰岛素释放,半定量RT-PCR检测CsA处理NIT-1胰岛β细胞后线粒体氧化磷酸化酶系中的几种主要成员(Nuox23、Cox7c和Atp5K)在mRNA水平上的表达。结果CsA处理24和48 h可显著抑制胰岛素的释放并下调Nuox23、Cox7c和Atp5K基因mRNA表达。结论CsA下调线粒体氧化磷酸化酶系基因的表达可能是其降低ATP合成引起胰岛素释放下降的机制之一。
Objective To investigate the effects of cyclosporin A on insulin release and the gene expression profiles of mitochondrial oxidative phosphorylation in NIT-1 cells. Methods NIT-1 cells were exposed to cyclosporin A (10 μmol/L) for 24 and 48 h respectively, after which the amount of insulin release was determined by means of radioimmunoassay (RIA), and the differential expressions of Nuox23, Cox7c and Atp5K genes assessed by semi-quantitative reverse transcription polymerase chain reaction. Results Cyclosporin A reduced insulin release in the cell culture after 24 and 48 h exposure and decreased Nuox23, Cox7c and Atp5K mRNA expressions. Conclusion Cyclosporin A induces inhibition of insulin release in NIT-1 cells, possibly due to the reduction of ATP synthesis involving the down-regulation of the gene expression of mitochondrial oxidative phosphorylation enzymes.
出处
《第一军医大学学报》
CSCD
北大核心
2003年第8期833-836,共4页
Journal of First Military Medical University
关键词
环孢霉素A
胰岛素
RT-PCR
NIT-1
细胞
cyclosporin A
insulin
reverse transcription polymerase chain reaction
NIT-1 cell line
