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�й�ҩѧ��־ 2013, Vol. 48 Issue (1) :13-16    DOI:
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DAI Yan-qing�� TU Mei-juan�� SUN Dong-li�� SHI Ai-qin�� ZENG Su�� JIANG Hui-di

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ժҪ Ŀ�� �����ȶ�����˹���ת����1��hPepT1������-���Ȯ��ϸ��ģ��(MDCK-hPepT1)��������Ӧ������ҩ�������˹���ת����1���Ƽ���ɸѡ������ ������������pcDNA3.1(+)-hPepT1������תȾ��-���Ȯ��ϵϸ������G418ɸѡ��������ϡ�ͷ���ѡ����¡ϸ�������˹���ת����1ӫ������-Ala-Lys(AMCA)�;������Glysar�Ե���¡ϸ��ת�˻��Խ�����֤��ͨ����ת¼�ۺ�ø��ʽ��Ӧ��֤�˹���ת����1��ת¼ˮƽ�ı�����ҩ�����Glysar��ȡ��Ӱ��ʵ���˹���ת����1���Ƽ���ɸѡ����� ��õ����굥��¡ϸ����D19��A11���Ԧ�-Ala-Lys(AMCA)��ȡ����������תȾ���������-���Ȯ��ϵϸ������Glysar����ȡ�ֱ�ΪתȾ������ϸ����24��11����D19��A11��hPepT1��mRNAˮƽ����������תȾ�������ϸ������֪���˹���ת����1���Ƽ�������������ٵ���¡ϸ��D19��A11��Glysar����ȡ��������ҩ�����Glysar��ȡ������ͬ�̶ȵ����ơ����� ��ʵ��ɹ��������ȶ��߱���˹���ת����1����-���Ȯ��ϵϸ��ģ�ͣ���Ӧ�ø�ģ��ɸѡ�����˹���ת����1����DZ���������õ���ҩ���塣
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Abstract�� OBJECTIVE To establish a cell model with stable-expressed human oligopeptide transporter 1(hPepT1)in MDCK�� then apply the cell model to screen inhibitors of hPepT1 from monomers of Chinese herbs. METHODS Recombinant plasmid pcDNA3.1(+)-hPepT1 was constructed and transfected into MDCK cells. After G418 screening�� monoclones were obtained by limiting dilution. Functional monoclones were selected and validated by uptake of fluorescent substrate ��-Ala-Lys(AMCA)and Glysar�� respectively�� while the reverse transcription-polymerase chain reaction(RT-PCR)was employed to validate the expression of hPepT1 at transcription level. Screening of hPepT1 inhibitors from monomers of Chinese herbs was achieved by investigating their effects on the uptake of Glysar. RESULTS Two monoclones named as D19 and A11 possessed significantly higher uptake of ��-Ala-Lys(AMCA)than that mock cells�� the uptake of Glysar in D19 and A11 were found to be increased up to 24 and 11-fold of mock cells�� respectively. The mRNA level of hPepT1 in D19 and A11 were significantly higher than that in mock cells�� the uptake of Glysar in D19 was obviously inhibited by known hPepT1 inhibitors(substrates). Some monomers from Chinese herbs inhibited the uptake of Glysar to different extent. CONCLUSION A cell model with stable-expressed hPepT1 is successfully established. Using this model�� a few of potential inhibitors of hPepT1 from monomers of Chinese herbs are found.
Keywords�� hPepT1,   MDCK,   cell model,   Chinese herbs,   monomer     
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ͨѶ���� ����淣�Ů������ �о�����ҩ��������л Tel����0571��88208408 E-mail��hdjiang@zju.edu.cn��HT4     Email: hdjiang@zju.edu.cn
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������, Ϳ����, �ﶬ��� .�ȶ�����˹���ת����1��ϸ��ģ�͹�����Ӧ��[J]  �й�ҩѧ��־, 2013,V48(1): 13-16
DAI Yan-Qing, TU Mei-Juan, SUN Dong-Li etc .Establishment and Application of Cell Model with Stable Expression of hPepT1[J]  Chinese Pharmaceutical Journal, 2013,V48(1): 13-16
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[1] BRANDSCH M. Transport of drugs by proton-coupled peptide transporters�� Pearls and pitfalls. Expert Opin Drug Metab Toxicol�� 2009�� 5(8)��887-905.
[2] GUO X J�� MENG Q�� LIU Q�� et al. Construction�� identification and application of HeLa cells stably transfected with human PEPT1 and PEPT2. Peptides�� 2012�� 34(2)��395-403.
[3] HERRERA-RUIZ D�� FARIA T N�� BHARDWAJ R K�� et al. A novel hPepT1 stably transfected cell line�� Establishing a correlation between expression and function. Mol Pharm�� 2004�� 1(2)��136-144.
[4] LU C�� LIAO M�� COHEN L�� et al. Emerging in vitro tools to evaluate cytochrome P450 and transporter-mediated drug-drug interactions. Curr Drug Discov Technol�� 2010�� 7(3)��199-222.
[5] CHU C�� OKAMOTO C T�� HAMM-ALVAREZ S F�� et al. Stable transfection of MDCK cells with epitope-tagged human PepT1. Pharm Res�� 2004�� 21(11)��1970-1973.
[6] ROMANO A�� BARCA A�� KOTTRA G�� et al. Functional expression of SLC15 peptide transporters in rat thyroid follicular cells. Mol Cell Endocrinol�� 2010�� 315(1-2)��174-181.
[7] SUN Y�� SUN J�� LIU J�� et al. Rapid and sensitive hydrophilic interaction chromatography/tandem mass spectrometry method for the determination of glycyl-sarcosine in cell homogenates. J Chromatogr B�� 2009�� 877(7)��649-652.
[8] DIECK S T�� HEUER H�� EHRCHEN J�� et al. The peptide transporter PepT2 is expressed in rat brain and mediates the accumulation of the fluorescent dipeptide derivative beta-Ala-Lys-Nepsilon-AMCA in astrocytes. Glia�� 1999�� 25(1)��10-20.
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[2] ������ ��־�� ��ΰ�� �޶� ��� ���� .��ЧҺ��ɫ�׼����ҩ������Ʒ����ӱ��ͱ��׵�8���򾲴���ҩ���о�[J]. �й�ҩѧ��־, 2012,47(6): 462-466
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[5] �ƾ�Ƽ����ԣ�������������ܣ�л��������ʢɽ.��ҩҩ��ѧ�о��и�ҩ��������״����[J]. �й�ҩѧ��־, 2012,47(21): 1685-1689
[6] Фѩ����濣������룬���������޹���.��ҩע������൰���ʳɷֲⶨ[J]. �й�ҩѧ��־, 2012,47(21): 1768-1769
[7] ��ϣ�����.���β���ҩ���ҩ���л����ѧ�о���չ[J]. �й�ҩѧ��־, 2012,47(18): 1430-1432
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[10] ������ � ������ ��Ӣ��.����Һ��ɫ��-�������׷��ⶨ��ʵ��ҩ���еĻ���ù����[J]. �й�ҩѧ��־, 2012,47(1): 65-68
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[13] Է��ͤ ���� ����ƽ ���л� ����.�ߴ�������Caco-2ϸ��ģ���е�ת�˻����о�[J]. �й�ҩѧ��־, 2011,46(4): 278-282
[14] ���� ���� ����� ���� ������ �ⰮӢ.UPLC-MS/MS�ⶨ��ҩ����19�ְ���������ũҩ������[J]. �й�ҩѧ��־, 2011,46(3): 235-236
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