HMGB1介导NF-kB通路促进多囊卵巢综合征胰岛素抵抗细胞模型的凋亡

摘要

Objective To investigate the effect of HMGB1 on IkB/NF-KB signaling pathway in ovarian granulosa cells of rats with insulin resistant. Methods The ovarian granulosa cells in vitro were cultured, and the control group, different concentrations of insulin intervention groups, different concentrations of HMGB1 intervention groups and insulin resistance+si-HMGB1 group and others were designed. The levels of glucose, HMGB1, androgen, TNF-α and phosphorylated NF-kB p65 were determined by enzyme-linked immuno sorbent assay (ELISA). The concentrations of IkBa, pIkBa, Caspase3 and Cleaved Casepase3 were detected by Western blotting. Results Compared with the control group, high insulin significantly decreased the glucose of rats (P<0.05), increased the androgen, HMGB1 and TNF-α (P<0.01) in ovarian granulosa cells, and then activated transcription factor NF-kB p65, significantly increased phosphorylated NF-kB p65(P<0.05). Androgen, phosphorylated IKBa, phosphorylated NF-kB p65 and TNF-a increased significantly with the increase of HMGB1 (P<0.05 or P<0.01). When adding HMGB1 interference small molecules into the ovarian granulosa cells, side effects induced by insulin resistance relieved, and the expression of androgen, Cleaved caspase 3, TNF-α, phosphorylated IkBa and phosphorylated NNF-kB p65(P<0.05 or P<0.01) were significantly lower than the control group. Conclusion HMGB1 could promote the increase of inflammatory factors in the PCOS insulin resistance ovarian granulosa cell model. HMGB1 may mediate the NF-kB signaling pathway, promote the cell apoptosis in insulin resistance ovarian granulosa cells, also involve in the pathogenesis of PCOS.%目的 探讨高迁移率族蛋白1(HMGB1)对胰岛素抵抗的大鼠卵巢颗粒细胞IkB/NF-kB信号通路的影响.方法 体外培养正常大鼠卵巢颗粒细胞,设对照组、不同浓度胰岛素干预组,不同浓度HMGB1干预组,胰岛素抵抗+si-HMGB1干预组等,用酶联免疫吸附法 (ELISA) 测定葡萄糖、HMGB1、雄激素、TNF-α、磷酸化NF-kB p65的含量;免疫印迹检测IkBa、pIkBa、Caspase3和Cleaved Caspase3的蛋白表达.结果 与对照组比较,胰岛素过高导致大鼠卵巢细胞的葡萄糖含量明显降低(P<0.05);雄激素及相关炎症因子HMGB1、TNF-α明显升高(P<0.01),并激活转录因子NF-kB p65并使其磷酸化程度明显升高 (P<0.05).随着HMGB1浓度升高,雄激素、磷酸化IkBa、磷酸化NF-kB p65和TNF-α的表达量较对照组均明显升高(P<0.05 or P<0.01);而当胰岛素抵抗的卵巢颗粒细胞中加入HMGB1的干扰小分子,则发现胰岛素抵抗带来的副作用得到缓解,较胰岛素抵抗对照组雄激素、Cleaved Cas-pase3、TNF-α、磷酸化IkBa和磷酸化NF-kB p65的表达量均显著性下降 (P<0.05 or P<0.01).结论 HMGB1在多囊卵巢综合征(PCOS)胰岛素抵抗卵巢颗粒细胞模型中促使炎症因子升高,可能介导了NF-kB信号通路的激活,促进胰岛素抵抗卵巢颗粒细胞凋亡,参与了PCOS的发病.