N-甲基芸香碱通过调控ERK通路诱导人胶质母细胞瘤细胞凋亡和自噬  

Induction of apoptosis and autophagy in human glioblastoma cells by N-methylflindersine:insights into regulatory role of ERK pathway

作　　者：黎思源 罗宇悠 罗雄明 王仲宇 陈晖彤 樊东 袁星怡 陈乐 唐佩 刘靖[1] 王宗明[2] 王欣 LI Siyuan;LUO Yuyou;LUO Xiongming;WANG Zhongyu;CHEN Huitong;FAN Dong;YUAN Xingyi;CHEN Le;TANG Pei;LIU Jing;WANG Zongming;WANG Xin(Department of Human Anatomy,Histology and Embryology,School of Basic Medical Sciences,Guangdong Pharmaceutical University,Guangzhou 510006,China;Pituitary Tumor Center,Department of Neurosurgery,the First Affiliated Hospital,Sun Yat-sen University,Guangzhou 510062,China;School of Life Sciences and Biopharmaceutics,Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广东药科大学基础医学院人体解剖学与组织胚胎学系,广东广州510006 [2]中山大学附属第一医院神经外科垂体瘤中心,广东广州510062 [3]广东药科大学生命科学与生物制药学院,药用生物活性物质研究所,广东广州510006

出　　处：《中国病理生理杂志》2024年第4期594-601,共8页Chinese Journal of Pathophysiology

基　　金：Supported by the National Natural Science Foundation of China for Young Scholars(No.81802678);the Guangdong Natural Science Foundation Project(No.2018A030310107);the Guangzhou Basic and Applied Basic Research Foundation(No.202102021116,No.202201010307,No.202201010299);the Guangdong Province Administration of Traditional Chinese Medicine Project(No.20231210).

摘　　要：目的:已知红树林相关植物能够产生具有抗肿瘤活性的天然化合物。尽管这些化合物具有潜在的治疗价值,但其抗肿瘤作用的分子机制尚需阐明。本研究侧重探讨一种来源于红树林相关植物大管[Micromelum falcatum(Lour.)Tan.]的生物碱N-甲基芸香碱对人胶质母细胞瘤U87细胞的影响。方法:从大管茎皮中分离得到15种化合物。其中,N-甲基芸香碱被筛选用于评估其对U87细胞生长的抑制作用。采用划痕实验、Hoechst 33342/PI染色和蛋白免疫印迹等多种方法,研究了该化合物对细胞迁移、凋亡以及与自噬相关的蛋白的影响。结果:N-甲基芸香碱在24 h内抑制了U87细胞的迁移活动,抗凋亡蛋白Bcl-2的表达减少,促凋亡蛋白Bax的表达增加,促使U87细胞发生凋亡,同时提高了U87细胞中自噬相关蛋白LC3-II/LC3-I的比值。此外,在N-甲基芸香碱处理细胞后,发现ERK信号通路被下调。结论:N-甲基芸香碱能够诱导U87细胞的凋亡和自噬性细胞死亡,从而抑制细胞生长,这种效应似乎与ERK信号通路有关。AIM:Mangrove-associated plants are known for producing natural compounds with antitumor activity.Despite the potential therapeutic value of these compunds,the molecular mechanisms underlying their antitumor effects remain unclear.This study aimed to investigate the antitumor properties of N-methylflindersine,an alkaloid derived from the mangrove-associated plant,Micromelum falcatum(Lour.)Tan.,and its effects on U87 human glioblastoma cells.METHODS:We identified and isolated 15 compounds from the stem bark of Micromelum falcatum.Among these,we screened N-methylflindersine for its potential inhibitory effects on U87 cell growth.Various assays,including wound healing,Hoechst 33342/PI staining,and protein expression analysis,were conducted to investigate the compound's impact on cell migration,apoptosis,and autophagy-related proteins.RESULTS:Within 24 h,N-methylflindersine demonstrated the ability to reduce U87 cell migration and increase the apoptotic U87 cell population.Furthermore,it downregulated the anti-apoptosis protein Bcl-2 expression,upregulated the pro-apoptosis protein Bax expression,and elevated the ratio of autophagy-related protein LC3-II/LC3-I in U87 cells.Additionally,the ERK signaling pathway was found to be down-regulated following N-methylflindersine treatment.CONCLUSION:N-methylflindersine appears to induce both apoptosis and autophagic cell death in U87 cells,resulting in reduced cell growth.This effect seems to be associated with the downregulation of the ERK signaling pathway.

关 键 词：红树林相关植物 N-甲基芸香碱 胶质母细胞瘤 

分 类 号：R739.4[医药卫生—肿瘤] R966[医药卫生—临床医学] R363.2