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作 者:胡灿[1] 李茹柳[1] 卢文彪[2] 羊燕群[1] 王静[1] 陈蔚文[1,3]
机构地区:[1]广州中医药大学脾胃研究所,广东广州510405 [2]广州中医药大学中药学院,广东广州510405 [3]上海市高校中医内科学E-研究院/上海中医药大学,上海201203
出 处:《中药材》2011年第5期738-746,共9页Journal of Chinese Medicinal Materials
基 金:国家自然科学基金(30772753);广东省自然科学基金(7300343);上海市教育委员会E-研究院建设计划项目(E03008)
摘 要:建立适合肠上皮整复研究的IEC-6细胞迁移药理实验模型,并观察黄芪糖复合物对细胞迁移的影响。方法:设立不同的细胞接种密度、划痕后观察时间、辅助材料Matrigel浓度、血清饥饿处理法、细胞迁移抑制剂DF-MO(二氟甲基鸟氨酸)浓度等以考察模型的建立条件,并在已建立的模型上观察受试药的效果。结果:①细胞宜以4×105/mL接种6孔板;②宜划痕后24 h观察细胞迁移数;③5%Matrigel为适宜浓度;④血清饥饿可造成细胞迁移数明显减少,应使用含血清的培养液。⑤DFMO 2.5~5 mmol/L为抑制细胞迁移适宜浓度。⑥黄芪糖复合物及阳性药精脒能促进细胞迁移。结论:建立了适宜药理实验的IEC-6细胞迁移模型,在此模型上可反映受试药对细胞正常迁移及迁移抑制的药效作用。Objective:To set up a suitable IEC-6 migration model for pharmacological research and observe the effect of complex polysaccharide from Astragalus membranaceus (Fisch.)Bge.var.mongholicus(Bge.) Hsiao to IEC-6 cell migration.Methods:The main conditions related to the establishment of the model,including the planting density of the cell,the observation time after scratching,the concentration of the auxiliary material Matrigel,the treatment of the serum-starvation,the concentration of α-difluoromethylornithine (DFMO),an inhibitor of the cell migration,were investigated respectively;and the effects of the tested medicines on the model were observed.Results:4×105cell/mL was the suitable planting density of the cell in the 6-well plate;at the 24th hour after scratching was the appropriate time to count the migrating cells;and the proper concentration of Matrigel was 5%;the serum-starvation could evidently reduce the migrating cells,so the culture medium should contain the serum;2.5 ~5 mmol/L DFMO was proper for inhibition of the cell migration.Complex polysaccharide fromAstragalus membranaceusand spermidine both can promote cell migration.Conclusion:The established model of IEC-6 cell migration was suitable for intestinal epithelial restitution such as the researches on pathophysiological mechanisms is the effects of the medicines on the cell migration.
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