9种中药提取物细胞毒性及其与人结肠腺癌细胞系单层细胞旁通透性的相关性研究  

Correlation of in vitro cytotoxicity with paracellular permeability in Caco-2 cells for the extracts of nine traditional Chinese medicine

作　　者：蔡润兰[1] 代勇[2] 王敏[1] 高源[1] 徐世军[2] 齐云[1] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]成都中医药大学,四川成都610075

出　　处：《中国药理学通报》2011年第10期1418-1421,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No81173645);国家"十一五"科技支撑计划资助项目(No2007BAI40B01)

摘　　要：目的了解9种中药提取物对Caco-2细胞毒性与对其细胞单层细胞-细胞间通透性的相关性。方法用MTT法测定9种中药提取物24 h及48 h的细胞毒,并计算IC50值。采用TranswellTM培养板构建Caco-2细胞单层,细胞电阻仪测定受试物对其TEER值的影响,找到影响细胞单层紧密连接的临界剂量,以此剂量与细胞毒参数行相关分析。结果 24、48 h细胞毒参数(IC50)与临界剂量(最大无毒剂量、最小有毒剂量)相关系数均>0.84,相关性检验P值均<0.01。最大无毒剂量与IC50的比值列阵显示,《小品方》甘草饮最小,三七醇提物最大。结论 9种中药提取物对Caco-2细胞毒与该细胞单层细胞旁通透性高度相关,采用细胞毒参数来预估中药提取物在Caco-2单层细胞模型供侧起始最大浓度具可行性。Aim To estimate the correlation of paracellular permeability（PP） with cytotoxicity in Caco-2 cell for the extracts of nine traditional Chinese medicine（TCM）.Methods Cytotoxicity was carried out using MTT cell viability assay in 96-well plates for 24 h or 48 h exposure time.PP was measured using TEER（membrane integrity indicator） on cells grown on inserts and the critical dose for effect on membrane integrity was analyzed.And correlation of cytotoxicity with the critical dose was investigated.Results The date showed that there were high correlations（r0.84） between IC50 values of the extracts for 24 h or 48 h exposure time and the critical dose（P0.01）.By arraying the ratios of the maximum nontoxic dose with IC50 values,the ratio of recipe of Gancaoyin was smallest and the extract of Panax notoginseng was biggest in the array.Conclusions The cytotoxicity displays the high correlation with PP in Caco-2 cells for the extracts of nine TCM.Therefore,it is feasible to depend on the IC50 values estimating initial maximum concentration of the extracts of TCM in donor compartment of Caco-2 cell monolayer model.

关 键 词：体外细胞毒 细胞旁通透性 CACO-2细胞 MTT活性测定 跨上皮电阻 相关性 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]