甘草酸二铵对雷公藤甲素诱导肠通透增强及染色体损伤的保护作用  

Protective effects of diammonium glycyrrhizinate on permeability increasing and chromosome breakage induced by triptolide

作　　者：淡墨[1] 文海若 闻镍[1] 刘丽[1] 李佐刚[1] 

机构地区：[1]中国食品药品检定研究院国家药物安全评价监测中心药物非临床安全评价研究北京市重点实验室,北京100176

出　　处：《中国新药杂志》2015年第7期750-754,770,共6页Chinese Journal of New Drugs

基　　金：国家"重大新药创制"科技重大专项(2012ZX09302001)

摘　　要：目的:研究雷公藤甲素(triptolide,TP)诱导的肠通透和染色体损伤及甘草酸二铵(diammonium glycyrrhizinate,DG)的保护作用。方法:首先利用Transwell培养体系,建立基于MDCK细胞系的肠通透模型,并通过测定细胞间隙标记物荧光素钠的通透速率对该模型的细胞间隙通透进行评价。利用CCK-8法测定DG对TP诱导MDCK细胞毒性的影响,并进一步观察TP对膜通透的影响及DG预处理24 h后的保护作用。MDCK经处理后接种于6孔细胞培养板,利用胞质分裂阻断法微核检测DG预处理对TP诱发的染色体损伤的影响。结果:荧光素钠表观渗透系数为(4.55±0.04)×10-8cm·s-1,MDCK培养模型形成良好的膜通透屏障。经CCK-8法测定,MDCK经TP孵育1,2和4 h时,IC50分别为40,23和20μmol·L-1。当MDCK经DG(12μmol·L-1)预处理24 h后,再与TP共同孵育1 h,IC50则升高至87μmol·L-1。此外,将TP(15μmol·L-1)加入MDCK模型受池孵育1 h可显著性增强MDCK细胞通透性,而DG(12μmol·L-1)24 h预处理可显著性降低TP对小肠通透性的影响。除细胞毒性,DG(12μmol·L-1)还可有效抑制由TP(15μmol·L-1)诱发的MDCK细胞微核率升高。结论:本研究提示DG可以显著性降低TP对MDCK的毒性及通透性。TP可通过诱发MDCK染色体断裂发挥遗传毒性效应,DG对TP诱导的MDCK染色体断裂亦有保护作用。Objective： To study the protective effects of diammonium glycyrrhizinate（ DG） on permeability and chromosome breakage induced by triptolide（ TP）. Methods： Transwell was used to establish the MDCK model. The paracellular permeability was measured using fluorescein sodium（ FS）. The effect of TP and the protective effect of DG on the paracellular pathway were studied. The cytotoxicity of TP and the protective effect of DG on MDCK were also investigated by CCK-8. The genotoxicity of TP and the protective effect of DG were also examined. Results： The MDCK model was established with Papp of FS（ 4. 55 ± 0. 04） × 10- 8cm·s- 1. After 1,2 and4 h treatment of TP,the IC50 s on MDCK were 40,23 and 20 μmol·L- 1,respectively. After 24 h pretreatment of DG,the IC50 on MDCK（ 1 h TP treatment） was increased to 87 μmol·L- 1. After 1 h TP treatment,the paracellular permeability of MDCK was significantly increased. After 24 h pretreatment of DG（ 12 μmol·L- 1）,the paracellular permeability of MDCK was significantly decreased compared with the TP（ 15 μmol·L- 1） group. The genotoxicities produced by 15 μmol·L- 1TP were attenuated by DG（ 12 μmol·L- 1） significantly. Conclusion： DG showed significant protective effects on the TP-induced cytotoxicity and MDCK permeability changes. TP may induce geno-toxicity on MDCK by promoting the chromosome breakage or loss,while DG showed potent protective effects against the genotoxicity.

关 键 词：雷公藤甲素 甘草酸二铵 小肠通透 染色体断裂 

分 类 号：R965.3[医药卫生—药理学]