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目的探讨同型半胱氨酸(Hcy)对克隆的胰岛β细胞NIT-1细胞株的存活率和凋亡的影响,以及对细胞内超氧化物歧化酶(SOD)的影响。方法将不同浓度的Hcy作用于NIT-1细胞后,用MTT的方法检测细胞生存率;用流式细胞仪Annexin V/PI双染色法和琼脂糖凝胶电泳检测不同浓度的Hcy作用不同时间对NIT-1细胞凋亡的影响;用黄嘌呤氧化酶法和WST结合的方法检测Hcy作用后的NIT-1细胞内SOD的活性。结果Hcy以时间、剂量依赖性的方式抑制NIT-1细胞的存活率(P<0.01)。Hcy可诱导NIT-1细胞的凋亡,随作用时间的延长和Hcy浓度增加NIT-1细胞的凋亡率逐渐增加,浓度为100μmol/L的Hcy作用24 h细胞凋亡明显增加,凋亡率为7.21%(P<0.01),250μmol/L的Hcy作用12 h后细胞凋亡率达8.91%(P<0.01)。100μmol/L的Hcy作用24 h后NIT-1细胞内SOD活性较正常组细胞下降20.2%(P<0.01)。结论Hcy可抑制NIT-1细胞的存活率,并以时间和剂量依赖性的方式诱导细胞凋亡;这些有害作用可能是通过抑制细胞内的SOD的活性而发挥作用,为研究保护胰岛细胞功能提供一种新的思路。
Objective To investigate the effect of homocysteine (Hcy) on the survival and apoptosis of cloned islet β cell NIT-1 cell line and the effect on the cell superoxide dismutase (SOD). Methods Different concentrations of Hcy were applied to NIT-1 cells and cell viability was detected by MTT assay. Flow Cytometry Annexin V / PI double staining and agarose gel electrophoresis were used to detect the effects of different concentrations of Hcy on NIT-1 cells. NIT-1 cells. The activity of SOD in NIT-1 cells treated with Hcy was detected by xanthine oxidase and WST. Results Hcy inhibited the survival of NIT-1 cells in a time-and dose-dependent manner (P <0.01). Hcy can induce the apoptosis of NIT-1 cells. The apoptosis rate of NIT-1 cells increased gradually with the prolongation of time and the increase of Hcy concentration. The apoptosis rate of NIT-1 cells increased obviously with Hcy concentration of 100μmol / L, Was 7.21% (P <0.01). After treated with 250μmol / L Hcy for 12 h, the apoptosis rate was 8.91% (P <0.01). The activity of SOD in NIT-1 cells was decreased by 20.2% (P <0.01) compared with normal cells after treated with 100μmol / L Hcy for 24 hours. Conclusion Hcy can inhibit the survival rate of NIT-1 cells and induce apoptosis in a time-and dose-dependent manner. These detrimental effects may play a role in inhibiting the activity of intracellular SOD, and provide the basis for studying the function of protecting islet cells A new way of thinking.