木犀草素通过增加ROS的产生和下调AKT/mTOR通路及HO-1蛋白表达抑制肺癌A549细胞增殖  

Luteolin inhibits proliferation of lung cancer A549 cells by increasing ROS production and inhibiting the AKT/mTOR signaling pathway and HO-1 expression

作　　者：李欢 邱紫欣 徐文洁 陈雪 魏典典 王允 LI Huan;QIU Zixin;XU Wenjie;CHEN Xue;WEI Diandian;WANG Yun(School of Public Health,Bengbu Medical University,Bengbu 233000,China)

机构地区：[1]蚌埠医科大学公共卫生学院,安徽蚌埠233000

出　　处：《南方医科大学学报》2024年第12期2367-2374,共8页Journal of Southern Medical University

基　　金：安徽省高校自然科学研究项目(2022AH051454,2022AH051425);蚌埠医学院自然科学研究项目(2021byzd030);蚌埠医学院研究生科研创新项目(Byycx22066)。

摘　　要：目的探究木犀草素(Lut)对肺癌A549细胞增殖的抑制作用及其内在机制。方法用不同浓度的Lut处理A549细胞48 h,通过MTT法检测细胞活性,通过平板克隆和EdU染色检测细胞增殖,通过DCFH-DA法检测细胞活性氧(ROS)水平,通过Hoechst33258染色法检测细胞凋亡水平,通过MDC染色法检测细胞自噬水平,通过Western blotting实验检测细胞凋亡相关蛋白Bax、Bcl-2、Cleaved caspase-9,自噬相关蛋白LC3B、Beclin1、P62,AKT/mTOR通路蛋白以及HO-1蛋白的表达。结果Lut剂量依赖性的抑制A549细胞的活力和增殖能力(P<0.05),引发细胞内ROS水平增加(P<0.05),上调凋亡相关蛋白Bax、Cleaved caspase-9和自噬相关蛋白Beclin1的表达,增加LC3B-II/LC3B-I的比值,下调抗凋亡蛋白Bcl-2和自噬相关蛋白P62的表达,诱导细胞凋亡和自噬(P<0.001)。此外,Lut可显著抑制AKT和mTOR的磷酸化,下调HO-1蛋白的表达(P<0.05)。结论Lut通过增加细胞内ROS的产生,抑制AKT/mTOR通路以及下调HO-1蛋白水平诱导A549细胞的凋亡和自噬。Objective To investigate the mechanism of luteolin for inhibiting proliferation of lung cancer A549 cells.Methods A549 cells treated with different concentrations of luteolin for 48 h were evaluated for changes in cell viability,proliferation,reactive oxygen species(ROS)production and apoptosis using MTT assay,plate cloning assay,EdU staining,DCFH-DA assay and Hoechst33258 staining.The changes in cell autophagy were examined with MDC staining,and the expressions of apoptosis-related proteins(Bax,Bcl-2,and cleaved caspase-9),autophagy-related proteins(LC3B,Beclin 1,and P62),AKT/mTOR pathway proteins,and HO-1 protein were detected using Western blotting.Results Treatment with luteolin dosedependently inhibited the viability and proliferation of A549 cells,increased intracellular ROS levels,up-regulated the expressions of Bax,cleaved caspase-9,and Beclin 1,increased the LC3B-II/LC3B-I ratio,down-regulated the expressions of Bcl-2 and P62,and induced cell apoptosis and autophagy.Luteolin also significantly inhibited the phosphorylation of AKT and mTOR and down-regulated the expression of HO-1 protein in the cells.Conclusion Luteolin induces apoptosis and autophagy to inhibit proliferation of A549 cells by increasing ROS production,inhibiting the AKT/mTOR pathway and downregulating HO-1 protein expression.

关 键 词：木犀草素 活性氧 凋亡 自噬 AKT/mTOR通路 HO-1 

分 类 号：R285[医药卫生—中药学]