蛋白酶体抑制剂Epoxomicin对视网膜色素上皮细胞自噬-溶酶体途径的影响

发布时间：2018-06-04 02:23

本文选题：泛素 + 蛋白酶体　；参考：《中山大学学报(医学科学版)》2016年04期

【摘要】：【目的】研究抑制泛素-蛋白酶体途径(UPP)对自噬-溶酶体途径(ALP)的影响及相关作用机制。【方法】人视网膜色素上皮细胞系ARPE-19共分为3组,分别为DMEM空白对照组、DMEM+DMSO阴性对照组、DMEM+Epoxomicin实验组。应用蛋白免疫印迹法和细胞免疫荧光法检测泛素化蛋白的表达,观察UPP功能的改变情况;观察LC3,LAMP1、LAMP2的表达,评价自噬作用和溶酶体的功能变化情况;观察HDAC6,p62的表达,初步探索UPP抑制对ALP改变的分子机制。【结果】采用蛋白酶体抑制剂Epoxomicin抑制泛素-蛋白酶体途径后,细胞内泛素化蛋白质聚集物增多;自噬溶酶体途径被激活,自噬标志物LC3表达增高,溶酶体膜相关蛋白LAMP1、LAMP2蛋白水平表达亦增高;抑制UPP后HDAC6和p62表达升高,可能参与ALP的激活过程。【结论】泛素-蛋白酶体途径抑制后,自噬溶酶体途径被激活且参与蛋白质聚集物的降解。
[Abstract]:[objective] to study the effect of UPP on autophagy lysosomal pathway (ALP) and its mechanism. [methods] Human retinal pigment epithelial cell line ARPE-19 was divided into three groups. DMEM Epoxomicin test group was used as DMEM blank control group and DMEM DMSO negative control group. The expression of ubiquitin protein was detected by Western blot and cell immunofluorescence to observe the change of UPP function, the expression of LC3 / LAMP1 / LAMP2 was observed to evaluate the changes of autophagy and lysosome function, and the expression of HDAC6 / p62 was observed. [results] after the proteasome inhibitor Epoxomicin was used to inhibit the ubiquitin proteasome pathway, the intracellular ubiquitin protein aggregates increased, and the autophagy lysosome pathway was activated. The expression of autophagy marker LC3 and lysosomal membrane-associated protein LAMP1 / LAMP2 was also increased, and the expression of HDAC6 and p62 was increased after UPP. [conclusion] Ubiquitin proteasome pathway inhibited the expression of HDAC6 and p62. [conclusion] Ubiquitin proteasome pathway inhibits the expression of HDAC6 and p62. Autophagy lysosome pathway is activated and involved in the degradation of protein aggregates.
【作者单位】：中山大学中山眼科中心//眼科学国家重点实验室;
【基金】：卫生部部属(管)医院2013-2015年度国家自然科学青年基金项目(81200670)
【分类号】：R774.1

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