基于小鼠胚胎干细胞实验模型评价川芎水煎液的胚胎毒性  

Evaluation of embryotoxicity of Chuanxiong Rhizoma decoction based on mouse embryonic stem cell test model

作　　者：夏荃 鲍倩[1] 蒋德菊[2] 易华[1] XIA Quan;BAO Qian;JIANG De-ju;YI Hua(Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Longgang Central Hospital of Shenzhen,Shenzhen 518116,China)

机构地区：[1]广州中医药大学,广东广州510006 [2]深圳市龙岗中心医院,广东深圳518116

出　　处：《中成药》2018年第9期1910-1915,共6页Chinese Traditional Patent Medicine

基　　金：广东省自然基金课题(2014A030313414)

摘　　要：目的建立小鼠胚胎干细胞实验模型,并以其评价川芎水煎液的胚胎毒性。方法体外培养小鼠胚胎成纤维细胞3T3和胚胎干细胞(mESCs)D3,MTT法检测青霉素G、苯妥英钠、5-氟尿嘧啶对3T3、D3细胞的毒性作用,绘制细胞存活率-浓度曲线,计算3种受试物对2种细胞的半数抑制浓度(IC_(50)3T3和IC_(50)D3)。利用悬滴-悬浮-贴壁方法,体外培养D3细胞分化成心肌细胞。通过实时定量PCR(Q-PCR)检测心肌细胞中肌球蛋白重链基因(β-MHC)的表达。再计算3种受试物对D3细胞的半数分化抑制浓度(ID50D3),判定其胚胎毒性等级。结果青霉素G、苯妥英钠、5-氟尿嘧啶分别为无、弱、强胚胎毒性。川芎水煎液的IC_(50)3T3、IC_(50)D3、ID50D3分别为9.39、18.78、10.20 mg/m L。结论川芎水煎液胚胎毒性较弱。AIM To establish mouse embryonic stem cell test model to evaluate the embryotoxicity of Chuan xiong Rhizoma decoction.METHODS Mouse embryonic fibroblasts 3T3 and embryonic stem cells(mESCs)D3 cultured in vitro were applied to testing the cytotoxic effects of penicillin G,phenytoin sodium and 5-fluorouracil on 3T3,D3 cells by MTT assay.According to drawn concentration-effect curves,the half inhibition concentrations of three test substances on two cells were calculated(IC503T3 and IC50D3).Myocardial cells cultured from D3 cells in vitro by hanging drop-suspension-attachment method.The expression of myosin heavy chain genes(β-MHC)in these myocardial cells were detected by real-time quantitative PCR(Q-PCR).The calculation of half inhibition cell differentiation concentrations(ID50 D3)of three test substances on D3 cells were subsequently performed,after which their embryotoxicity grades were determined.RESULTS Penicillin G was identified with no embryotoxicity,phenytoin sodium was found to be with weak embryotoxicity,and 5-fluorouracil was strongly embryotoxic.The IC503T3,IC50D3,ID50D3 of Chuanxiong Rhizoma decoction were determined to be 9.39,18.78,10.20 mg/mL,respectively.CONCLUSION Chuanxiong Rhizoma decoction demonstrates a weak embryotoxicity.

关 键 词：川芎水煎液 胚胎毒性 胚胎干细胞实验模型 

分 类 号：R285.5[医药卫生—中药学]