白藜芦醇对镉诱导HEK 293T细胞自噬与凋亡的影响  

The Effects of Resveratrol on the Relationship between Cadmium-Induced Autophagy and Apoptosis

作　　者：王芸芸 刘小綪 邵峦峦 杨颖 蒋琦辰[1] 毛伟平[1] WANG Yunyun;LIU Xiaoqian;SHAO Luanluan;YANG Ying;JIANG Qichen;MAO Weiping(Jiangsu Province Key Laboratory for Molecular and Medicine Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing Jiangsu 210046, China)

机构地区：[1]南京师范大学生命科学学院江苏省分子医学重点实验室,江苏南京210046

出　　处：《复旦学报（自然科学版）》2022年第1期52-60,共9页Journal of Fudan University：Natural Science

摘　　要：为了探究白藜芦醇与镉诱导人胚胎肾细胞293(HEK 293T)细胞自噬与凋亡之间的关系,本研究在已证明低浓度镉诱导细胞自噬,高浓度镉导致细胞凋亡的基础上,以HEK 293T细胞为研究对象,并分别与镉、白藜芦醇及各种抑制剂进行联合培养,利用免疫印记分析检测自噬标志蛋白LC3B-Ⅱ/Ⅰ、溶酶体标志蛋白cathepsin L、组蛋白去乙酰化酶相关蛋白SIRT1和凋亡蛋白caspase-3的表达水平,激光共聚焦检测细胞自噬质粒(GFP-LC3)和溶酶体相关膜蛋白结合因子(LAMP2)的表达量,同时检测活性氧(ROS)水平,倒置相差显微镜和流式细胞术检测细胞凋亡情况。结果显示:白藜芦醇进一步增强镉诱导HEK 293T细胞的自噬,且这种效应发生在自噬体溶酶体融合之前,其作用原理可能与白藜芦醇的抗氧化性有关并与SIRT1基因有密切联系,以上结果说明白藜芦醇对镉诱导的细胞自噬有一定的调控作用,此外,细胞凋亡与自噬之间关系紧密,白藜芦醇可抑制镉诱导的细胞凋亡。To explore the relationship between the effect of resveratrol on cadmium-induced autophagy and apoptosis in HEK 293T cells,based on the evidence that low concentrations of cadmium induce cell autophagy and high concentrations of cadmium lead to apoptosis,HEK 293T cells were cultured in combination with cadmium and resveratrol,then western blot analysis was used to detect the expression levels of autophagy marker protein(LC3B-Ⅱ/Ⅰ),lysozyme marker protein(cathepsin L),silent information regulation 2 homolog 1(SIRT1)and apoptosis protein(caspase-3).Laser confocal was used to detect the expression of pcDNA3.1-GFP-LC3B(GFP-LC3)and 1ysosomal-associated membrane protein(LAMP2),while detecting reactive oxygen species(ROS)levels,inverted phase contrast microscopy was used.Flow cytometry was used to detect apoptosis.The results showed that resveratrol could enhance cadmium-induced HEK 293T cell autophagy,and this effect occured before autophagy lysosome fusion.The mechanism may be related to the antioxidant properties of resveratrol and closely related to the SIRT1 gene.The above results showed that resveratrol has a regulatory effect on cadmium-induced cell autophagy.In addition,there is a strong relationship between apoptosis and autophagy,and resveratrol inhibits cadmium-induced apoptosis.

关 键 词：白藜芦醇 镉 HEK 293T细胞 自噬 细胞凋亡 

分 类 号：R965.2[医药卫生—药理学] R992[医药卫生—药学]