预知子经ROS介导的PI3K-Akt通路诱导非小细胞肺癌细胞凋亡  

Fructus Akebiae induces apoptosis via regulating ROS-mediated PI3KAkt signaling pathway in non-small cell lung cancer cells

作　　者：高万里 周绮纯 王苏美 GAO Wanli;ZHOU Qichun;WANG Sumei(Clinical and Basic Research Team of Traditional Chinese Medicine Prevention and Treatment of Non-Small Cell Lung Cancer,Second Clinical College of Guangzhou University of Chinese Medicine,Guangzhou 510520,Guangdong,China)

机构地区：[1]广州中医药大学第二临床医学院中医药防治非小细胞肺癌临床与基础研究团队,广东广州510520

出　　处：《中国临床药理学与治疗学》2025年第3期339-346,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金面上项目(82274602)。

摘　　要：目的:探讨预知子诱导人非小细胞肺癌(NSCLC)细胞死亡方式及机制。方法:通过CCK-8实验、Hochest33342/PI染色实验以及克隆形成实验检测预知子对细胞生存、增殖能力的影响,采用Western blot检测预知子干预后对NSCLC细胞内死亡相关蛋白ATG5、HMGB1、GPX4,凋亡蛋白及PI3K-Akt信号通路相关蛋白的表达变化,采用流式细胞术检测细胞凋亡率,通过DCFH-DA荧光探针法观察检测细胞内活性氧(ROS)水平。结果:预知子能明显抑制NSCLC细胞的存活率和增殖能力(P<0.05),预知子处理后NSCLC细胞内凋亡相关蛋白BAX、Cleaved caspase3和Cleaved caspase9表达显著上调(P<0.05),p-PI3K、p-Akt和BCL2蛋白表达均明显下调(P<0.05),预知子可明显促进NSCLC细胞的凋亡率和细胞内ROS量升高(P<0.05),抗氧化剂NAC能逆转预知子导致的NSCLC细胞凋亡、ROS水平升高及PI3K-AKT通路蛋白的调控作用(P<0.05)。结论:预知子通过ROS介导的PI3K-Akt通路导致了NSCLC细胞凋亡。AIM:To explore the way and mechanism of cell death induced by Fructus Akebiae in non-small cell lung cancer(NSCLC)cells.METHODS:CCK-8,Hochest33342/PI staining,and colony formation experiments were used to detect cell viability and proliferation.Western blot was used to detect the expression of cell death-related proteins ATG5,HMGB1,GPX4,apoptosis-related proteins,and PI3K-Akt signaling pathway-related proteins.Flow cytometry was used to detect the apoptosis rate.DCFH-DA fluorescent probes were used to detect the levels of total intracellular ROS under a fluorescence microscope.RESULTS:Fructus Akebiae significantly inhibited the cell viability and proliferation of human NSCLC cells(P<0.05).The expression of apoptosis-related proteins BAX,Cleaved caspase3,and Cleaved caspase9 in NSCLC cells was significantly increased(P<0.05),while p-PI3K,p-Akt,and BCL2 were markedly decreased(P<0.05).Fructus Akebiae was found to clearly increase the apoptosis rate and the levels of total intracellular ROS(P<0.05).The antioxidant NAC significantly reversed apoptosis,ROS production and regulation of PI3KAkt pathway related proteins induced by Fructus Akebiae(P<0.05).CONCLUSION:Fructus Akebiae induces apoptosis via regulating ROS-mediated PI3KAkt signaling pathway in non-small cell lung cancer cells.

关 键 词：预知子 非小细胞肺癌 凋亡 PI3K-Akt通路 ROS 

分 类 号：R285[医药卫生—中药学] R730.5[医药卫生—中医学] R734.2