理中汤、人参皂苷Rg1、6-姜酚对Caco-2细胞PepT1转运功能影响的比较

Comparison of Lizhong Decoction,Ginsenoside Rg1 and 6-Gingerol on the Transport Ability of PepT1 in Caco-2 Cells

目的:观察比较理中汤、人参皂苷Rg1、6-姜酚对Caco-2细胞肽转运载体PepT1转运二肽的能力及其蛋白、mRNA表达的影响。方法:Caco-2细胞融合后连续培养28d后分别给予理中汤、人参皂苷Rg1、6-姜酚处理,并设立正常对照组及cAMP抑制剂对照组,用放射性同位素示踪技术比较Caco-2细胞转运二肽化合物Glycyl-Sarcosine的能力,采用westernblot方法测定Caco-2细胞膜上PepT1蛋白的表达,荧光定量PCR方法测定PepT1mRNA(SLC15A1)的表达水平。结果:理中汤、人参皂苷Rg1及6-姜酚均在不同程度上表现为促进Caco-2细胞转运Glycyl-Sarcosine的作用;Caco-2细胞经理中汤/人参皂苷Rg1/6-姜酚处理24h后,细胞膜PepT1蛋白表达水平均明显增高。6-姜酚能明显上调Caco-2细胞SLC15A1表达,人参皂苷对Caco-2细胞SLC15A1表达的作用表现为下调,理中汤对Caco-2细胞SLC15A1表达作用不明显。且理中汤、人参皂苷Rg1、6-姜酚三者作用各有特点。结论:理中汤及其成分人参皂苷Rg1、6-姜酚具有促进正常培养Caco-2细胞转运二肽化合物Glycyl-Sarcosine的作用,该过程调控与胞内第二信使cAMP有一定的关系。

Objective：The aims of this study were to observe the effects of 6-gingerol on expression and transport ability of PepT1 in Caco-2 cells.Methods：Caco2 monolayers were grown on permeable supports.Peptide transport activity was studied using-glycyl-sarcosine（-Gly-Sar）.The densities of PepT1 protein and mRNA（SLC15A1）expression levels were analyzed by Western blot and Real-time quantitative Polymerase Chain Reaction.Results：The total transported Gly-Sar of Caco-2 cells treated by Lizhong Decoction,Ginsenoside Rg1 or 6-Gingerol were higher than those of controlled group.And they showed up-regulated effect on PepT1 protein expression.6-gingerol can up-regulate the mRNA expression.Conclusion：Lizhong Decoction and its compounds ginsenoside Rg1 and 6-gingerol have significant effects on promoting the transport ability of dipeptides in Caco-2 cells,which maybe partly regulated by cAMP pathway.