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目的研究核转录因子(NF-κB)在高浓度葡萄糖诱导INS-1细胞凋亡中的作用。方法大鼠胰岛素瘤细胞系(INS-1)细胞以RPMI1640培养基常规培养。实验分对照组(11.1mmol/l葡萄糖组)、高糖组(33.3mmol/l葡萄糖组)、高糖+NF-κB抑制剂组(33.3mmol/l葡萄糖+NF-κB抑制剂(5μmol/l)干预组)3组。以荧光定量RT-PCR检测IKKβ mRNA水平,Western blot法检测细胞核内NF-κB亚基P65蛋白表达量,Annexin V-PI双染法检测细胞凋亡率。结果与对照组相比较,高糖组IKKβ mRNA水平显著升高(P<0.01),INS-1细胞胞核内P65蛋白表达显著增多(P<0.01),细胞凋亡率显著升高(P<0.05)。与高糖相比较,高糖+NF-κB抑制剂组IKKβ mRNA水平显著下降(P<0.01),胞核内P65蛋白表达显著减少(P<0.01),INS-1细胞凋亡率显著下降(P<0.05)。结论高浓度葡萄糖诱导INS-1细胞NF-κB活化,抑制NF-κB活化可有效抑制高糖诱导的INS-1细胞凋亡。
Objective To investigate the role of nuclear factor-kappa B (NF-κB) in the apoptosis of INS-1 cells induced by high glucose. Methods The rat insulinoma cell line (INS-1) cells were routinely cultured in RPMI1640 medium. The experiment was divided into control group (11.1mmol / l glucose group), high glucose group (33.3mmol / l glucose group), high glucose + NF-κB inhibitor group (33.3mmol / l glucose + NF-κB inhibitor ) Intervention group) 3 groups. The level of IKKβ mRNA was detected by real-time RT-PCR. The expression of P65 protein in NF-κB subunit was detected by Western blot. The apoptosis rate was detected by Annexin V-PI double staining. Results Compared with the control group, the level of IKKβ mRNA in high glucose group was significantly increased (P <0.01), the expression of P65 protein in INS-1 cells was significantly increased (P <0.01), and the apoptosis rate was significantly increased (P < 0.05). Compared with high glucose, the level of IKKβ mRNA was significantly decreased in high glucose + NF-κB inhibitor group (P <0.01), the expression of P65 protein in nucleus was significantly decreased (P <0.01), and the apoptosis rate of INS-1 cells was significantly decreased P <0.05). Conclusion High glucose can induce the activation of NF-κB in INS-1 cells and inhibit the activation of NF-κB in INS-1 cells.