苦楝素诱导小鼠肝细胞DNA损伤  

Toosendanin induced DNA damage in mouse hepatocytes

作　　者：卢艳琳 叶杰[1] 闫晓凤 张宇佳 王晓玲[1] LU Yan-lin;YE Jie;YAN Xiao-feng;WANG Xiao-ling(Department of biology,School of basic medicine,Shanghai University of traditional Chinese medicine(Shanghai,201203)China)

机构地区：[1]上海中医药大学基础医学院生物教研室,上海201203 [2]上海中医药大学附属龙华医院肿瘤科

出　　处：《中西医结合肝病杂志》2020年第4期330-332,共3页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：国家自然科学基金项目(No.81703832);十三五“重大新药创制”科技重大专项(No.2017ZX09304001);龙华医院国家中医临床研究基地“龙医团队、龙医学者育苗”项目(No.LYTD-65)。

摘　　要：目的:观察苦楝素对DNA损伤及损伤修复的影响。方法:体外采用苦楝素与小鼠胚胎肝细胞BNL CL2细胞株共培养,CCK-8检测肝细胞活力,吸光度检测肝细胞内ROS含量,Western blot检测γ-H2AX、Parp-1、poly ADP-ribose蛋白表达,观察不同浓度苦楝素以及苦楝素不同作用时间对DNA损伤及损伤修复的影响。结果:随着苦楝素浓度升高,肝细胞BNL.CL2细胞活力逐渐下降(P<0.0001),ROS含量升高(P<0.05),同时γ-H2AX蛋白质含量随着苦楝素作用时间的延长而升高(P<0.0001)。苦楝素作用0.5~1.0 h Parp-1自身PAR化量达最高(P<0.05);而苦楝素作用时间自1~24 h Parp-1的PAR逐渐减少(P<0.05)。结论:苦楝素可通过增加细胞内ROS诱导DNA损伤,同时抑制Parp-1的活性从而抑制DNA损伤的修复。Objective:To observe the effect of Toosendanin on DNA damage and repair.Methods:Toosendanin was cultured with BNL CL2 cell line.To observe the effect of different concentration and different time of action of Toosendanin on DNA damage and repair,CCK-8 was used to detect the hepatocyte viability and ROS was used to detect absorbance of ROS content in hepatocytes.Western blot was used to detect the expression ofγ-H2AX,Parp-1,poly ADP-ribose.Results:With the concentration of Toosendanin increased,the viability of BNLCL2 cells was decreased gradually(P<0.0001)and the ROS content was increased(P<0.05).At the same time,the protein content ofγ-H2AX was increased with the prolongation of action time of Toosendanin(P<0.0001)and the protein content of poly ADP-ribose was the highest in 0.5-1 hour(P<0.05).But the protein content of poly ADP-ribose was decreased gradually from 1 hour to 24 hours(P<0.05).Conclusion:Toosendanin can inhibit the repair of DNA damage by increasing ROS induced DNA damage and inhibiting Parp-1 activity.

关 键 词：肝细胞 苦楝素 DNA损伤 

分 类 号：R735.7[医药卫生—肿瘤]