摘要
目的利用HepaRG细胞,探讨补骨脂二氢黄酮甲醚毒性剂量及其毒性作用机制。方法 MTT法确定毒性及剂量。以浓度为6.25、12.5、25μmol·L^(-1)的补骨脂二氢黄酮甲醚作用于细胞24 h后,首先利用Hoechst 33342/PI及LDH的方法检测细胞死亡,并检测凋亡相关蛋白Bax、Bcl-2、caspase-3活性。通过检测线粒体膜电位、ATP含量、线粒体膜通道开放孔开放性及细胞色素C的含量,观察其对线粒体损伤的影响。结果补骨脂二氢黄酮甲醚给药24 h后,凋亡蛋白Bax/Bcl-2比例、caspase-3活性随药物浓度增大而增加,在6.25~25μmol·L^(-1)时,其凋亡率呈现明显的剂量依赖性,但在25μmol·L^(-1)时细胞以坏死为主。作用24 h后线粒体损伤相关指标明显加剧。结论补骨脂二氢黄酮甲醚作用于HepaRG细胞后通过损伤线粒体诱导细胞凋亡和坏死。
Aim To probe into the bavachinin toxic mechanisms in HepaRG cells.Methods MTT assay was used to determine toxicity and dose.After treated with 6.25,12.5 and 25μmol·L-1 bavachinin for 24 h,the cell death was investigated by Hoechst 33342/PI,LDH,caspase-3 activity,and Bax,Bcl-2 expression levels.Mitochondrial damage was detected by mitochondrial membrane potential,ATP content,openness of mitochondrial permeability transition pore and cytochrome C level.Results Bavachinin administration up-regulated Bax/Bcl-2 ratio and caspase-3 activity with increase of drug concentration after 24 h.Apoptotic rate increased in a dose-dependent manner between 6.25 and 25μmol·L-1,but when reached to 25μmol·L-1,cells showed more necrosis.Mitochondrial injury indicators significantly increased after 24h.Conclusion Bavachinin induces HepaRG cell apoptosis and necrosis through mitochondria injury.
作者
季宇彬
王敏
王姗
罗云
孙桂波
孙晓波
JI Yu-bin;WANG Min;WANG Shan;LUO Yun;SUN Gui-bo;SUN Xiao-bo(Research Center on Life Sciences and Environmental Sciences,Harbin University of Commerce,Harbin 150076,China;Pharmacology and Toxicology Center,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences,Beijing 100193,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2018年第4期544-550,共7页
Chinese Pharmacological Bulletin
基金
国家科技部"重大新药创制"科技重大专项(No2015ZX09501004-001-003)
中国医学科学院医学与健康科技创新工程(No 2017-12M-1-013)
