阿司匹林对胰岛瘤细胞株NIT-1细胞增殖及NF-κB表达的影响

Effect of aspirin on proliferation and nuclear factor kappa B p65 expression in NIT-1 insulinoma cells

目的探讨阿司匹林对胰岛瘤细胞株NIT-1细胞增殖、胰岛素分泌及NF-κB p65的影响。方法培养胰岛细胞株NIT-1细胞,经不同浓度阿司匹林处理后,采用MTT法检测细胞增殖,Hoechest33342荧光染色法观察细胞核形态和细胞凋亡变化,用放射免疫法测定胰岛素分泌和通过NF-κB p65激活-核转运试剂盒在荧光显微镜下检测NF-κB p65的表达。结果随药物浓度增加和作用时间的延长,阿司匹林对NIT-1细胞增殖的抑制作用逐渐增强,细胞培养24h时,5mmol/L阿司匹林组与对照(NC)组比较(P=0.011374),10mmol/L阿司匹林组与NC组比较(P=0.000079),差异均有统计学意义。随阿司匹林剂量的不断增加,细胞培养各时间段均出现不同的剂量-效应关系。当药物浓度在1mmol/L时,细胞培养48h后,细胞增殖明显变缓慢,胰岛素分泌水平降低,且表现出明显的时间-效应关系。随阿司匹林浓度的增加,凋亡细胞数明显增多,细胞内NF-κB p65表达显著下降。结论阿司匹林对胰岛瘤细胞株NIT-1的增殖具有抑制作用,其抑制作用随阿司匹林浓度增加,呈时间-效应和剂量-效应关系。

Objective To investigate the effect of aspirin on proliferation, insulin secretion, and nuclear factor kappa B （NF-κB） p65 protein expression in insulinoma NIT-1 cells. Methods The cultured insulinoma NIT-1 ceils were treated with aspirin in different concentrations. Effect of aspirin on proliferation of NIT-1 ceils was evaluated by MTT assay, and the morphological and apoptotic changes were observed by immunofluorescence techniques with Hoechst 33342 staining. The insulin was measured by insulin Radioimmunoassay detection Kit. Expression of NF-κB p65 was determined under a fluorescence microscope by Cellular NF-κB Translocation Kit. Results Inhibition rate of the NIT-1 cells was obviously enhanced along with the increased aspirin concentratiorL When cultured for 24 h, in the comparisons of 5mmol/L aspirin group with the control group （P = 0. 011374） and 10mmol/L aspirin group with the control group （P=0. 000079）, the differences were statistically significant. Along with the increase of aspirin doses, there appeared the dose-dependent changes of NIT-1 cell proliferation and NF-κB p65 expression. When the cells cultured in aspirin at 1 mmol/L for 48 h, the cell proliferation became slow significantly, and the insulin secretion was lowered, showed a significant time-effect relationship. When the aspirin dose increased, the apoptotic cells were getting significantly more, while the expression of NF- κB p65 was getting significantly less. Conclusion Aspirin can inhibit the proliferation of pancreatic islet NIT-1 cells, and this inhibition shows time-effect and dose-effect relations with the increase of aspirin concentration.