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目的探讨葡萄糖、肿瘤坏死因子(TNF)-α、干扰素(IFN)-γ对胰岛细胞自身抗原胰岛素瘤相关蛋白(IA)-2表达水平及胰岛素分泌水平的影响。方法以不同浓度的葡萄糖(3、7、11、30mmol/L)、10ng/mlIFN-γ及100ng/mlTNF-α分别刺激胰岛细胞系RIN5F24h和48h,用反转录聚合酶链反应(RT-PCR)方法检测各组RIN5F细胞中IA-2的mRNA表达水平,并用放射免疫法检测上清中胰岛素的浓度。结果RT-PCR结果显示以不同浓度葡萄糖(3、7、11、30mmol/L)分别刺激RIN5F细胞系后IA-2mRNA水平呈浓度依赖性和时间依赖性增加;TNF-α可抑制IA-2mRNA的水平,并且作用48h后IA-2表达水平显著降低(P<0.05);IFN-γ作用24h及48h均可显著抑制IA-2的表达(P<0.05)。TNF-α或IFN-γ刺激RIN5F细胞系后,上清中胰岛素水平均较基础值降低,并且作用48h后差异有统计学意义(P<0.05)。结论葡萄糖可使IA-2的mRNA水平呈剂量依赖性和时间依赖性增加,TNF-α、IFN-γ可抑制IA-2的mRNA表达和胰岛素分泌浓度。
Objective To investigate the effect of glucose, tumor necrosis factor (TNF) -α and interferon-γ (IFN-γ) on the expression of insulinoma-associated protein (IA-2) and insulin secretion in pancreatic islets. Methods The islet cell line RIN5F was stimulated with different concentrations of glucose (3, 7, 11, 30mmol / L), 10ng / ml IFN- γ and 100ng / ml TNF- α for 24 hours and 48 hours, respectively. Reverse transcriptase-polymerase chain reaction ) Method was used to detect the mRNA expression of IA-2 in RIN5F cells in each group, and the concentration of insulin in the supernatant was detected by radioimmunoassay. Results The results of RT-PCR showed that the concentration of IA-2 mRNA in RIN5F cell line stimulated by different concentrations of glucose (3, 7, 11 and 30mmol / L) increased in a concentration-dependent and time-dependent manner. TNF- (P <0.05). IA-2 expression was significantly inhibited by IFN-γ at 24h and 48h (P <0.05). After RIN5F cell line was stimulated by TNF-|Á or IFN-|Ã, the level of insulin in the supernatant was lower than the baseline, and the difference was statistically significant after 48h (P <0.05). Conclusion Glucose can increase mRNA level of IA-2 in a dose-dependent and time-dependent manner. TNF-α and IFN-γ can inhibit IA-2 mRNA expression and insulin secretion.