理中汤对Caco-2细胞PepT1转运功能的影响  

Effects of Lizhong Decoction on the 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10期108-111,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:观察理中汤对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后给予理中汤处理,并设立空白及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,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:理中汤处理组Caco-2细胞15,30,60,120 min时点Glycyl-Sarcosine吸收转运累计量分别为(4.52±0.57),(6.34±0.57),(7.29±0.64),(8.23±0.67)μmol/孔,高于空白对照组(P<0.05),后者分别为(2.42±0.68),(3.15±0.76),(5.62±0.20),(6.54±0.54)μmol/孔;理中汤与Rp-8-Br-cAMP合用组15,30,120 min时点分别为(2.68±0.46),(4.13±0.46),(6.73±0.43)μmol/孔,明显低于相应时点单用理中汤组(P<0.05);理中汤处理后Caco-2细胞膜PepT1蛋白表达增加,PepT1表达与内参GAPDH表达灰度比值理中汤组为(0.326±0.031),高于空白对照组(0.220±0.019)(P<0.01);理中汤对Caco-2细胞SLC15A1 mRNA表达未见明显作用,但与cAMP抑制剂合用,理中汤能逆转Rp-8-Br-cAMP下调Caco-2细胞SLC15A1 mRNA表达的作用。结论:理中汤具有促进正常培养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 effects of Lizhong Decoction on expression and transport ability of PepT1 in Caco-2 cells. Method： Caco2 monolayers were grown on permeable supports. Peptide transport activity was studied using [ ^14C ]-glycyl-sarcosine （[^ 14C ]-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 15, 30, 60, 120 min of group treated by Lizhong Decoction were higher than those of controlled group, （4.52 ± 0.57 ）, （6.34 ± 0.57 ）, （7.29 ± 0.64）, （ 8.23 ± 0. 67 ） μmol/well vs （2.42 ±0.68）, （3.15 ±0.76）, （5.62 ±0.20）, （6.54 ±0.54） μmol/well. The group of Lizhong Decoction together with Rp-8-Br-cAMP at 15, 30, 120 min. were 2.68 ±0.46,4. 13 ±0.46,6.73 ±0.43 μmol/p well. The figures were significantly lower than that of Lizhong Decoction olone. The PepT1 protein expression level of cells treated with Lizhong Decoction was higher that of control. Conelusion：Lizhong Decoction has significant effects on promoting the transport ability of dipeptides in Caco-2 cells, which maybe take effects by inducing the plasmatic PepT1 protein to insert to cell membrane.

关 键 词：理中汤 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]