人参皂苷Rg1对Caco-2细胞PepT1转运功能的影响  

Effect of Ginsenoside Rg1 on Transport Ability of PepT1 in Caco-2 Cells

作　　者：郭文峰[1] 胡灿[1] 温鹏[1] 高小玲[1] 李茹柳[1] 陈蔚文[1,2] 

机构地区：[1]广州中医药大学脾胃研究所,广东广州510405 [2]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：《广州中医药大学学报》2011年第4期388-392,共5页Journal of Guangzhou University of Traditional Chinese Medicine

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

摘　　要：【目的】观察人参皂苷Rg1对Caco-2细胞肽转运载体PepT1转运二肽化合物的能力及其蛋白、mRNA表达的影响。【方法】Caco-2细胞长满瓶底后继续培养28 d,然后给予人参皂苷Rg1(终浓度为50 mmol/L)处理,并设立空白对照组及cAMP抑制剂(Rp-8-Br-cAMP)对照组,采用放射性同位素示踪技术比较Caco-2细胞对二肽化合物Glycyl-Sarcosine的转运能力,采用Western blot法测定Caco-2细胞膜上PepT1蛋白的表达,荧光定量PCR法测定PepT1 mRNA(SLC15A1)的表达水平。【结果】人参皂苷Rg1处理组Caco-2细胞15、30、60、120 min时点Glycyl-Sarcosine吸收转运累计量均显著高于空白对照组对应时点的量(P<0.05或P<0.01),人参皂苷Rg1与Rp-8-Br-cAMP合用组15、30、60、120 min时点均显著低于相应时点单用人参皂苷Rg1组(P<0.05或P<0.01),人参皂苷Rg1处理后Caco-2细胞膜PepT1蛋白表达显著增加(P<0.05);荧光定量PCR结果显示:人参皂苷Rg1能显著下调SLC15A1表达(P<0.05)。【结论】人参皂苷Rg1具有促进正常培养Caco-2细胞转运二肽化合物Glycyl-Sarcosine的作用,该作用可能与其促进胞浆中PepT1蛋白嵌入胞膜中发挥转运二肽的功能有关,该过程调控与胞内第二信使cAMP有一定的关系。Objective To observe the effect of ginsenoside Rg1 on transport ability of peptide transport carrier PepT1 for bipeptid compound and on protein and mRNA expression of PepT1 in Caco-2 cells.Methods Caco-2 monolayers were grown on permeable supports for 28 days.Then the cultured Caco-2 cells were treated with ginsenoside Rg1 or cyclic adenosine monophosphate（cAMP） inhibitor（Rp-8-Br-cAMP）.Peptide transport activity was studied by using glycyl-sarcosine.PepT1 protein expression was detected by Western blot method,and PepT1 mRNA（SLC15A1） expression level was analyzed by real-time fluorescent quantitative polymerase chain reaction（PCR）.Results At time points of 15,30,60 and 120 min,the glycyl-sarcosine amount conveyed by Caco-2 cells was higher in ginsenoside Rg1 group than that in the blank control group（P0.05 or P0.01）,and was lower in the combination group of ginsenoside Rg1 and Rp-8-Br-cAMP than that in ginsenoside Rg1 group（P0.05 or P0.01）.After treatment with ginsenoside Rg1,the protein expression of PepT1 was increased and SLC15A1 expression level was down-regulated（P0.05）.Conclusion Ginsenoside Rg1 can promote the transport of glycyl-sarcosine in normal Caco-2 cells,and the mechanism is probably related with the enhancement of cytoplasm PepT1 inserting cell membrane and with the regulation of intracellular cAMP.

关 键 词：人参皂苷/药理学 基因表达调控 转运功能 细胞培养 

分 类 号：R285.5[医药卫生—中药学]