胰高血糖素样肽-1对AGEs诱导H9C2心肌细胞自噬的影响

发布时间：2018-07-01 14:04

  本文选题：胰高血糖素样肽素- + 糖基化终产物　； 参考：《中国药理学通报》2017年03期

【摘要】：目的观察胰高血糖素样肽-1(GLP-1)对晚期糖基化终产物(AGEs)诱导H9C2心肌细胞自噬(Autophagy)的影响,初步探讨GLP-1心肌细胞保护作用与自噬活性关系。方法将传代培养H9C2心肌细胞随机分4组:(1)Control组:加入0.9%生理盐水;(2)AGEs组:加入100 mg·L~(-1)AGEs;(3)AGEs+GLP-1组:同时加入100 mg·L~(-1)AGEs和10 nmol·L~(-1)GLP-1;(4)AGEs+GLP-1+Rapamycin组:同时加入100 mg·L~(-1)AGEs、10 nmol·L~(-1)GLP-1及5μmol·L~(-1)Rapamycin(自噬诱导剂)。各组在经上述预处理24 h后,分别用CCK-8法检测细胞存活率Hoechst 33258试剂盒检测细胞的凋亡率,DCFH-DA荧光探针检测细胞内活性氧(ROS)水平,流式细胞术检测细胞自噬溶酶体形成,Western blot法检测自噬相关蛋白(LC3Ⅱ,Beclin-1)表达。结果 (1)与Control组相比,AGEs组细胞生存率明显减少,细胞内ROS水平明显增高,细胞自噬溶酶体水平及自噬相关蛋白(LC3Ⅱ、Beclin-1)表达均明显增加;(2)与AGEs组相比,AGEs+GLP-1组细胞生存率明显增高,细胞内ROS水平、细胞自噬溶酶体水平及自噬相关蛋白(LC3Ⅱ、Beclin-1)表达均明显下降;(3)与AGEs组相比,AGEs+GLP-1+rapamycin组中细胞内ROS水平降低,而细胞生存率、细胞自噬溶酶体水平及自噬相关蛋白(LC3Ⅱ、Beclin-1)表达未见差异。结论 (1)AGEs可导致H9C2心肌细胞内ROS升高,诱导细胞损伤并激活细胞自噬;(2)GLP-1对AGEs诱导的H9C2心肌细胞损伤具有保护作用,其保护机制可能与抑制细胞自噬活性有关。
[Abstract]:Objective to observe the effect of glucagon like peptide 1 (GLP-1) on autophagy (Autophagy) induced by advanced glycation end products (ages), and to explore the relationship between the protective effect of GLP-1 and autophagy activity. Methods cultured H9C2 cardiomyocytes were randomly divided into four groups: (1) Control group: 0. 9% saline; (2) ages group: 100 mg L ~ (-1) ages; (3) ages GLP-1 group: 100 mg L ~ (-1) ages and 10 nmol L ~ (-1) GLP-1; (4) ages GLP-1 Rapamycin group: 100 mg L ~ (-1) ages 10 nmol L ~ (-1) GLP-1 and 5 渭 mol L ~ (-1) rapamycin. After pretreatment for 24 h, the rate of apoptosis and the level of reactive oxygen species (Ros) were detected by CCK-8 Hoechst 33258 kit and DCFH-DA fluorescence probe, respectively. Flow cytometry was used to detect the expression of autophagy associated protein (LC3 鈪，

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