6-姜酚对Caco-2细胞PepT1转运功能的影响  

Effects of 6-gingerol on Transport Ability of PepT1 in Caco-2 Cells

作　　者：郭文峰[1] 杨伟鹏[2] 王怡薇[2] 王彦礼[2] 胡灿[1] 温鹏[1] 高小玲[1] 陈蔚文[1,3] 

机构地区：[1]广州中医药大学脾胃研究所,广州510405 [2]中国中医科学院中药研究所,北京100700 [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：《中国实验方剂学杂志》2011年第16期116-119,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(30600798);上海市教育委员会E-研究院建设计划项目(E03008)

摘　　要：目的:观察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28 d后给予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Western blot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PepT1 mRNA(SLC15A1)的表达水平。结果:6-姜酚处理组Caco-2细胞60,120 min时点Glycyl-Sarcosine吸收转运累计量分别为(6.84±0.46),(8.61±0.54)μmol/孔,高于正常对照组(P<0.05),后者分别为(5.62±0.20)、(6.54±0.54)μmol/孔;6-姜酚与Rp-8-Br-cAMP合用组120 min时点为(6.92±0.67)μmol/孔,明显低于相应时点单用6-姜酚组(P<0.05);6-姜酚处理后Caco-2细胞膜PepT1蛋白表达增加,PepT1表达与内参GAPDH表达灰度比值6-姜酚组为(0.317±0.022),高于空白对照组(0.220±0.019)(P<0.01);荧光定量PCR结果,6-姜酚能上调Caco-2细胞SLC15A1 mRNA表达,但与cAMP抑制剂合用后该抑制作用被阻断。结论: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,该作用可能与上调PepT1蛋白及mRNA表达有关,该过程调控与胞内第二信使cAMP有一定的关系。Objective： To observe the effects of 6-gingerol on expression and transport ability of PepT1 in Caco-2 cells.Method： Caco-2 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.Result： The total transported Gly-Sar of Caco-2 cells within 60,120 minutes of group treated by 6-gingerol were higher than those of controlled group,（6.84±0.46）,（8.61±0.54） μmol/per well vs（5.62±0.20）,（6.54±0.54） μmol/per well,and higher than group of cells treated together with Rp-8-Br-cAMP within 120 minutes,the latter result were（6.92±0.67） μmol/per well.And 6-gingerol showed up-regulated effect on PepT1 protein and mRNA expression,also can be inhabited by Rp-8-Br-cAMP.Conclusion： 6-gingerol has significant effects on promoting the transport ability of dipeptides in Caco-2 cells,which maybe take effects by up-regulating PepT1 protein and mRNA expression.

关 键 词：6-姜酚 肽转运载体1 转运功能 环磷酸腺苷 

分 类 号：R285.5[医药卫生—中药学]