曲古霉素A和5-氮杂胞苷对细胞因子诱导损伤的胰岛β细胞的保护作用

Protective effect of trichostatin a and 5-azacitidine on cytokine-induced toxicity in pancreatic β-cells

目的 探讨细胞因子白细胞介素-1β和干扰素-γ诱导下曲古霉素A(TSA)、5-氮杂胞苷(5-AzaC)及其联合干预对胰岛p细胞增殖、凋亡及功能的影响.方法 采用白细胞介素-1β和干扰素-γ诱导大鼠胰岛素瘤细胞(RIN-m5f)损伤,用TSA、5-AzaC对其进行干预.具体分组:空白对照组、细胞因子诱导组、0.05/0.10 μmol/L TSA干预组、0.63/1.25 μmol/L 5-AzaC干预组、0.10 μmol/L TSA+ 1.25 μmol/L5-AzaC联合干预组.用MTT法检测RIN-m5f细胞的增殖活性,流式细胞技术检测细胞凋亡率及酶联免疫吸附法测定葡萄糖刺激胰岛素分泌能力.结果 0.05/0.10 μmol/L TSA干预组、0.63/1.25μmol/L 5-AzaC干预组、0.10 μmol/L TSA+ 1.25tμmol/L 5-AzaC联合干预组细胞增殖活性分别为70.1％/79.2％、67.3％/82.9％和89.1％,高于细胞因子诱导组的33.9％(P＜0.05);其凋亡率分别为10.3％/10.5％、7.9％/9.6％和8.2％,低于细胞因子诱导组的16.6％(P＜0.05);各组葡萄糖刺激胰岛素分泌能力均比细胞因子诱导组升高(P＜0.05).结论 TSA和5-AzaC可以促进细胞因子诱导的胰岛β细胞增殖,抑制其凋亡和恢复p细胞的胰岛素分泌功能.

Objective To investigate the effect of trichostatin A (TSA) and 5-azacitidine (5-AzaC) on pancreatic β-cells impaired by cytokine,via measuring the proliferation,apoptosis,and function of pancreatic β-cells.Methods RIN-m5f was impaired by interleukin-1β and interferon-γin vitro,and treated with TSA and 5-AzaC.Experiment groups included blank control group,cytokine induction group,0.05/0.10 μmoL/L TSA group,0.63/1.25 μmoL/L 5-AzaC group,and0.10 μmol/L TSA plus 1.25 μmol/L 5-AzaC group.The viability of RIN-m5f cells was detected by MTT assay.Apoptotic rate was determined by Annexin V-fluorescein isothiocyanate (FITC) /propidium iodide flow cytometry.Insulin secretion was measured by enzyme-linked immunosorbent assay.Results The viability of RIN-m5f cells in 0.05/0.10 μmoL/L TSA group,0.63/1.25 μmol/L 5-AzaC group,and 5-AzaC plus TSA group was 70.1％/79.2 ％,67.3 ％/82.9 ％,and 89.1％ respectively,being higher than that in the cytokine group (33.9％,P＜0.05) ; the apoptosis rate was 10.3％/10.5％,7.9％/9.6％,and 8.2％,being lower than that in the cytokine group (16.6％,P＜0.05) ; the capacity of glucose-stimulated insulin secretion of all the treated groups was higher than that in the cytokine group (P＜0.05).Conclusion TSA and 5-AzaC might promote the proliferation of pancreatic β-cells impaired by cytokines,inhibit its apoptosis and recover its insulin secretion.