褪黑素对软脂酸诱导L02细胞胰岛素抵抗的防护作用及机制

发布时间：2018-06-22 22:41

本文选题：胰岛素抵抗 + 活性氧ROS　；参考：《中国老年学杂志》2017年13期

【摘要】：目的探讨褪黑素(MT)对软脂酸(PA)诱导L02细胞胰岛素抵抗的防护作用及机制。方法培养L02细胞,分别设立对照组(BSA组)、PA组、PA+MT组、PA+维生素C(Vit C)组。各组药物浓度和孵育时间根据不同的实验需要来确定。用荧光比色法测定各组细胞胞内氧自由基(ROS)水平,Western印迹法检测胰岛素刺激后胞内胰岛素信号蛋白磷酸化水平,包括Y-p-胰岛素受体(IR)、Y-p-胰岛素受体底物(IRS)1/2以及应激敏感激酶p-JNK水平。结果用不同浓度的PA处理L02细胞24 h,细胞内ROS生成量呈现明显的浓度依赖关系,差异有统计学意义(P0.05);用0.3 mmol/L的PA处理L02细胞不同时间,细胞内ROS生成量存在明显的时间依赖关系,在24 h达到最高,并在36 h后仍维持较高水平(P0.05);与BSA组相比,PA组ROS含量明显升高(P0.05)PA+MT组ROS含量无明显变化(P0.05);PA+MT组ROS生成量明显低于PA组(P0.05)。PA+Vit C组与PA组相比,ROS的生成量减少,但仍高于PA+MT组(P0.05)。与BSA组相比,PA组Y-p-IR、Y-p-IRS1/2磷酸化水平减少,而p-JNK磷酸化水平升高;与PA组相比,PA+MT组Y-p-IR、Y-p-IRS1/2磷酸化水平显著升高,p-JNK磷酸化水平明显降低(P0.05)。结论 PA导致细胞内ROS增加,MT能够清除PA产生的过量ROS。MT通过减少JNK的活化,改善因ROS引起的胰岛素信号传递损伤,对PA诱导的胰岛素抵抗具有防护作用。
[Abstract]:Objective to investigate the protective effect and mechanism of melatonin (MT) on insulin resistance induced by palmitate (PA) in L02 cells. Methods L02 cells were cultured and divided into two groups: control group (BSA group), PA group (PA MT group) and vitamin C (Vit C) group. Drug concentration and incubation time were determined according to different experimental needs. Intracellular oxygen free radical (Ros) levels were measured by fluorescence colorimetry and phosphorylation of insulin signaling proteins was detected by Western blotting, including 1 / 2 of Y-p- insulin receptor (IR) -Y-p- insulin receptor substrate (IRS) and stress sensitive kinase (p-JNK). Results when L02 cells were treated with different concentrations of PA for 24 h, the intracellular Ros production showed a significant concentration-dependent relationship (P0.05), and 0.3 mmol / L PA was used to treat L02 cells for different time. The intracellular Ros production was time-dependent and reached its highest level at 24 h. After 36 hours, the Ros content in PA group was significantly higher than that in BSA group (P0.05). The Ros production in PA MT group was significantly lower than that in PA group (P0.05), but still higher than that in PA MT group (P0.05). Compared with BSA group, the phosphorylation level of Y-p-IRN Y-p-IRS1 / 2 decreased and p-JNK phosphorylation increased in PA group, and the phosphorylation level of Y-p-IRY-p-IRS1 / 2 in PA MT group was significantly higher than that in PA group (P0.05). Conclusion PA can decrease the activation of JNK and improve the insulin signal transduction injury induced by Ros, which can protect against the insulin resistance induced by PA.
【作者单位】：河南科技大学医学院生物化学与分子生物学教研室;天津市东丽区中医医院外科;
【基金】：河南科技大学创新能力培育基金(No.2012ZCX024)
【分类号】：R587.1

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