槟榔碱通过氧化应激介导的内皮间充质转化致HUVE细胞损伤的机制研究  

Damage mechanism of arecoline on HUVECs triggered by oxidative stress-mediated endothelial-to-mesenchymal transition

作　　者：邓国磊 何双桃[1] 李国良[1] 朱可可 DENG Guolei;HE Shuangtao;LI Guoliang;ZHU Keke(Southern Branch of the First People's Hospital of Chenzhou,Chenzhou,423000,Hunan,China;The First Hospital of Hunan University of Chinese Medicine,Changsha,410007,Hunan,China)

机构地区：[1]郴州市第一人民医院南院,湖南郴州423000 [2]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《肿瘤药学》2022年第4期488-495,共8页Anti-Tumor Pharmacy

基　　金：国家自然科学基金(81874496);湖南省中医药管理局青年项目(No.2021240)。

摘　　要：目的基于氧化应激介导的内皮间充质转化(EndMT)探讨槟榔碱(Arecoline)诱导人脐静脉内皮细胞(HUVEC)损伤的作用机制研究。方法采用槟榔碱干预复制HUVECs损伤模型。实验设对照组、槟榔碱高剂量组和槟榔碱低剂量组。采用CCK-8检测不同浓度槟榔碱对HUVECs存活率的影响,细胞成像分析检测HUVECs形态学变化,采用MitoSOX探针检测线粒体活性氧(ROS)水平,采用免疫荧光及激光共聚焦检测氧化应激相关蛋白PGC-1α、Nrf2及EndMT相关蛋白E-cadherin、N-cadherin、Vimentin、SNAI1的表达情况,Western blotting检测氧化应激相关蛋白PGC-1α、Nrf2及EndMT相关蛋白N-cadherin、Vimentin的表达情况。结果槟榔碱可以浓度依赖性抑制HUVECs活性,降低细胞存活率,诱导细胞形态结构发生变化,明显下调PGC-1α、Nrf2蛋白的表达,促进细胞线粒体释放ROS,同时显著下调细胞中E-cadherin蛋白的表达,上调N-cadherin、Vimentin和SNAI1蛋白的表达,最终诱导HUVECs发生EndMT,显著加重细胞损伤。结论槟榔碱对HUVECs具有明显的损伤作用,可能是通过氧化应激介导的EndMT发挥作用,这可能是槟榔碱所致口腔黏膜下纤维化病变继而诱发口腔癌的重要病理机制。Objective To investigate the damage mechanism of arecoline on the human umbilical vein endothelial cells(HUVEC) triggered by oxidative stress-mediated endothelial-to-mesenchymal transition(EndMT).Methods The HUVECs model induced by arecoline was established according to literature.The cultured cells were randomly divided into control group,arecoline high-dose group and arecoline low-dose group.The viability of cells treated by different concentrations of arecoline was detected by CCK-8 assay.Morphological change of HUVECs was observed by cell imaging analysis.Mitochondrial reactive oxygen species(ROS) was analyzed by MitoSOX fluorescent probe.The expressions of oxidative stressrelated proteins PGC-1α and Nrf2,and EndMT-related proteins E-cadherin,N-cadherin,Vimentin and SNAI1 were detected by immunofluorescent staining and laser scanning confocal microscope.The expressions of oxidative stress-related proteins PGC-1α,Nrf2 and EndMT-related proteins N-cadherin,Vimentin were also detected by Western blotting.Results Arecoline could significantly inhibit the viability of HUVECs by a concentration-dependent way.Additionally,it could induce morphological changes in HUVECs,and observably accelerate abnormal increase of mitochondrial ROS.Furthermore,it could obviously down-regulate the expressions of PGC-1α,Nrf2 and E-cadherin,while up-regulate the expressions of N-cadherin,Vimentin and SNAI1.It eventually induced EndMT in HUVECs,and significantly aggravated the damage of HUVECs.Conclusion Arecoline exhibits remarkable damage effects on HUVECs,which may be triggered by oxidative stress-mediated EndMT.This might be the important pathological mechanism for oral submucous fibrosis-mediated oral carcinoma induced by arecoline.

关 键 词：槟榔碱 氧化应激 内皮间充质转化 口腔黏膜下纤维化 口腔癌 

分 类 号：R739.85[医药卫生—肿瘤] R285.5[医药卫生—临床医学]