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目的 研究转录因子NF-κB在细胞因子[白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和干扰素γ(IFN-γ)]诱导的INS-1胰岛β细胞损害中的作用。方法 构建携有NF-κB的抑制物IκBα的突变体IκBα△N的逆转录病毒,采用免疫印迹分析、κB报告基因荧光分析、四甲基偶氮唑盐细胞活力分析等方法进行研究。结果 IL-1β可诱导INS-1细胞的IκBα降解和NF-κB激活,而对感染IκBα△N逆转录病毒的INS-1/IκBα△N细胞则无此作用。IL-1β加IFN-γ或TNF-α加IFN-γ可致INS-1细胞活力降低,而INS-1/IκBα△N细胞可抵抗这些细胞因子导致的细胞损害。结论IL-1B、TNF-α可导致β细胞活力降低,IFN-γ可能起促进作用,抑制NF-κB活性可能保护β细胞免于上述细胞因子介导的损害。
Objective To investigate the role of transcription factor NF-κB in INS-1 islet β-cell damage induced by cytokines [IL-1β, TNF-α and IFN-γ] Role. Methods The retrovirus carrying IκBα △ N mutant of IκBα, an inhibitor of NF-κB, was constructed and analyzed by immunoblot analysis, fluorescence analysis of κB reporter gene and cell viability analysis of tetramethylzarbazole salt. Results IL-1β induced IκBα degradation and NF-κB activation in INS-1 cells, but no effect on INS-1 / IκBαΔN cells infected with IκBαΔN retrovirus. IL-1β plus IFN-γ or TNF-α plus IFN-γ resulted in decreased viability of INS-1 cells, whereas INS-1 / IκBαΔN cells were able to counteract the cellular damage caused by these cytokines. Conclusion IL-1B and TNF-α can induce the decrease of β-cell viability and the enhancement of IFN-γ. Inhibition of NF-κB activity may protect β-cells from cytokine-mediated damage.