加味丹玄口康对槟榔碱诱导的HUVECs细胞内皮间充质转化的影响  

Effect of Jiawei Danxuan Koukang on arecoline-induced endothelial to mesenchymal transition of HUVECs cells

作　　者：朱可可 任振虎[2,3] 谭怡丝 谢圣发 谭劲 朱萱[5] ZHU Ke-ke;REN Zhen-hu;TAN Yi-si;XIE Sheng-fa;TAN Jin;ZHU Xuan(First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Ninth Hospital of Shanghai Jiao Tong University School of Medcine,Shanghai 200011,China;National Center for Stomatology/National Clinical Research Center for Oral Disease,Shanghai 200011,China;Medicine Xintian County Hospital of Traditional Chinese Medicine,Xintian Hunan 425799,China;Hunan University of Chinese Medicine,Changsha Hunan 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]上海交通大学医学院附属第九人民医院,上海200011 [3]国家口腔医学中心/国家口腔疾病临床医学研究中心,上海200011 [4]新田县中医院,湖南新田425799 [5]湖南中医药大学,湖南长沙410208

出　　处：《时珍国医国药》2023年第3期572-576,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(81874496);湖南省中医药管理局重点项目(201808);湖南省中医药管理局优秀青年项目(2021240);湖南省教育厅科研项目(21B0398,22C0190);湖南省长沙市指导性科技计划(kzd2001010)。

摘　　要：目的基于氧化应激介导的内皮间充质转化(EndMT)探讨加味丹玄口康对槟榔碱诱导所致HUVECs细胞损伤的保护机制。方法参照文献方法复制槟榔碱诱导的人脐静脉内皮细胞(HUVECs)损伤模型,实验设正常组、模型组、空白血清对照组和加味丹玄口康含药血清组。采用CCK-8法检测不同浓度的槟榔碱及加味丹玄口康含药血清对HUVECs细胞存活率的影响,细胞成像分析检测各组HUVECs细胞形态学变化,采用MitoSOX荧光探针检测线粒体活性氧(ROS),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及间充质转化标志物相关蛋白E-cadherin、N-cadherin、Vimentin、SNAI1表达情况。结果加味丹玄口康能有效改善槟榔碱诱导所致HUVECs细胞损伤,提高细胞存活率(P<0.01),明显抑制细胞线粒体活性氧(ROS)异常升高(P<0.01),显著逆转氧化应激相关蛋白PGC-1α、Nrf2表达减少(P<0.01或P<0.05),同时上调细胞EndMT相关蛋白E-cadherin、下调EndMT相关蛋白N-cadherin、Vimentin和SNAI1表达(P<0.01或P<0.05)。结论加味丹玄口康对槟榔碱诱导所致HUVECs细胞损伤具有的保护作用,其是通过抑制氧化应激介导的EndMT进而发挥抗槟榔碱损伤作用,这可能是加味丹玄口康抗槟榔碱所致的口腔黏膜下纤维化病变继而预防口腔癌的重要机制。Objective To investigate the protective mechanism of Jiawei Danxuan Koukang against damage of arecoline-induced HUVECs cells based on endothelial to mesenchymal transition mediated by oxidative stress.Methods Cell model of HUVECs in-duced by arecoline was replicated according to literature.The cultured cells or rats were randomly divided into normal group,model group,blank serum group,and Jiawei Danxuan Koukang group.The HUVECs cell viability treated by several concentra-tions of arecoline and jiawei Danxuan Koukang containing drug serum was detected by MTT assay.Morphological changes of HU-VECs cells was detected by cell imaging analysis.Mitochondrial reactive oxygen species(ROS)was analyzed by MitoSOX fluo-rescent probe.The oxidative stress-related protein expression of PGC-1α,Nrf2 and EndMT-related protein expression of E-cadherin,N-cadherin,Vimentin and SNAI1were test by immunofluorescent staining and laser scanning confocal microscope.Results Jiawei Danxuan Koukang can significantly alleviate damage of HUVECs cells induced by arecoline,increase cell survival(P<0.01),and markedly inhibit abnormal increasement of mitochondrial reactive oxygen species(ROS).Moreover,Jiawei Danxuan Koukang can also obviously reverse the down-regulated expression of oxidative stress-related proteins PGC-1αand Nrf2(P<0.01 or P<0.05).Additionally,the endMT-related protein expression of E-cadherin was up-regulated and that the expression of E-cadherin,N-cadherin,Vimentin and SNAI1 was down-regulated by Jiawei Danxuan Koukang treatment(P<0.01or P<0.05).Conclusion Jiawei Danxuan Koukang has remarkable protective effects against damage of arecoline-in-duced HUVECs cells,via restraining EndMT mediated by oxidative stress,which may be the important mechanism of Jiawei Danxuan Koukang for the prevention and treatment of oral submucous fibrosis(OSF)induced by arecoline.

关 键 词：加味丹玄口康 槟榔碱 氧化应激 内皮间充质转化 细胞损伤 

分 类 号：R285.5[医药卫生—中药学]