中国药理学与毒理学杂志2012，Vol.26Issue(6)：864-869,6.DOI:10.3867/j.issn.1000-3002.2012.06.015
基于胚胎干细胞实验模型评价黄芩苷的胚胎毒性
Evaluation of embryotoxicity of baicalin based on embryonic stem cell test system
摘要
Abstract
OBJECTIVE To assess embryotoxicity of baicalin using embryonic stem ( ES) cell test in vitro. METHODS ES D3 cells and BALB/c 3T3 cells were cultured respectively with baicalin 20, 40, 60, 80 and 100 mg·L-1 . Cell morphology was observed with a phase contrast microscope and absorbance of the resulting colored solution at 570 nm was measured by microplate reader. According to the concentration-effect curve, 50% inhibition of cell growth ( IC50 ) and viability were obtained in ES cell line D3 [ IC50 (D3) ] and in 3T3 cells [ IC50 (3T3 ) ]. ES cells were cultured in baicalin with using hanging drop-suspension-attachment method, then cardiac myoblasts specific genes myosin heavy chain ((3-MHC) in differentiation of embryonic stem cell were detected by real time Q-PCR, the growth inhibitory rate was calculated with quantitative analysis, according to the concentration-effect curve, 50% inhibition of ES cells differentiation into cardiac myoblasts [ ID50 ( D3 ) } obtained. Baicalin embryotoxicity potential was predicted using statistics formula. RESULTS ES D3 cells and BALB/c 3T3 cells viability decreased slowly with the increase in baicalin concentration after they were cultured in different concentration of baicalin for ten days, which showed that baicalin had a certain degree of inhibition on ES D3 cell and BALB/c 3T3 cell proliferation. The half-maximal proliferation-inhibition concentration (IC50 ) of baicalin on ES cell line D3 [IC50(D3)j and 3T3 cells[IC50(3T3) ] was 135.9 mg·L-1 and 63.34 mg·L-1. ES cells in vitro by hanging drop-suspension-adherent culture could differentiate to expression of β-MHC gene in myocardial cells. With baicalin 2, 5, 10, 20 and 40 mg·L-1 , the ability of ES cells differentiate into myocardial cells gradually decreased, the inhibitory rate was 29. 5% , 46. 8% , 59. 6% , 61. 7% and 69. 0% , respectively, and this indicated that baicalin had a certain degree of inhibitory effect on ES cells differentiation. The half-maximal differentiation-inhibition concentration(ID50) of baicalin on D3 differentiating into MHC (ID50 (D3) ] was 7. 25 mg · L -1. According to the calculation formula of predicting embryo toxicity, baicalin had weak embryotoxicity. CONCLUSION Baicalin has weak embryotoxicity.关键词
胚胎干细胞/黄芩苷/毒性作用Key words
embryonic stem cell/ baicalin/ toxic actions分类
医药卫生引用本文复制引用
张崴,宋殿荣,王雅楠,朱泽..基于胚胎干细胞实验模型评价黄芩苷的胚胎毒性[J].中国药理学与毒理学杂志,2012,26(6):864-869,6.基金项目
国家中医药管理局中医药行业科研专项(200707011) （200707011）
