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作 者:郑倩 王晓珊 林蔓婷 沈思霞 朱思婷 宋冠华 ZHENG Qian;WANG Xiaoshan;LIN Manting;SHEN Sixia;ZHU Siting;SONG Guanhua(School of Life Science,Huizhou University,Huizhou 516000,Guangdong,China)
出 处:《惠州学院学报》2023年第3期35-39,共5页Journal of Huizhou University
基 金:广东省普通高校特色创新类项目(2019KTSCX179);广东省科技创新战略专项“攀登计划”(pdjh2022b0500);广东省大学生创新创业训练项目(S202110577059)。
摘 要:为探究异乌药内酯诱导细胞内活性氧的产生在介导线粒体凋亡途径中的作用及对肺癌A549细胞凋亡的影响,将A549细胞分为对照组(DMSO)、异乌药内酯处理组(15μmol/L)、抗氧化剂NAC组(3 mmol/L)和异乌药内酯联合NAC处理组(Isolinderalactone 15μmol/L+NAC 3 mmol/L)进行实验。采用CCK-8方法检测细胞活性,通过流式细胞术检测细胞凋亡情况,DCFH-DA法检测A549细胞内活性氧水平变化,JC-1染色后流式细胞术检测细胞内线粒体膜电位,Western blot方法检测细胞内线粒体凋亡通路相关关键蛋白表达的变化。结果表明:异乌药内酯能抑制肺癌A549细胞生长并诱导细胞凋亡,促进ROS积累,线粒体膜显著去极化,上调促凋亡蛋白Bax表达水平而下调抗凋亡蛋白Bcl-2表达水平。抗氧化剂NAC与异乌药内酯联合作用后,与异乌药内酯单独处理组相比,以上各方面的变化水平都呈现出不同程度的下降。异乌药内酯能通过诱导细胞内活性氧积累,激活线粒体凋亡途径从而促进A549细胞发生凋亡。To investigate the role of reactive oxygen species(ROS)accumulation induced by Isolinderalactone in mitochondrial apop‐totic pathway and the effect on apoptosis in A549 cells,the A549 cells were divided into control group(DMSO),Isolinderalactone group(15μmol/L),antioxidant NAC group(3 mmol/L)and Isolinderalactone+NAC group(Isolinderalactone 15μmol/L+NAC 3 mmol/L)for assays.CCK-8 assay was employed to detect the cell viability.Flow cytometry after DCFH-DA fluorescence staining was used to eval‐uated the level of ROS in A549 cells.JC-1 fluorescence staining was to detect the intracellular mitochondrial membrane potential(MMP).Western blot was applied to measure the expression levels of key protein associated with mitochondrial apoptotic pathway in A549 cells.Isolinderalactone exhibited a significant inhibitory effect on the viability of A549 cells,induced cell apoptosis,increased the intracellular ROS level,depolarized MMP,promoted the expression level of pro-apoptotic protein Bax and down-regulated the expres‐sion of anti-apoptotic protein Bcl-2.After pretreatment with antioxidant NAC,the viability of A549 cells was increased,and the rate of cell apoptosis was dramatically decreased.Furthermore,the level of increased intracellular ROS,depolarized MMP and ratio of Bax/Bcl2 were all significantly decreased.Isolinderalactone induced A549 cell apoptosis by promoting intracellular ROS accumulation to acti‐vate mitochondrial apoptotic pathway.
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