Nitric oxide mediates increased P-glycoprotein activity in interferon-γ-stimulated human intestinal cells

被引:21
作者
Dixit, SG
Zingarelli, B
Buckley, DJ
Buckley, AR
Pauletti, GM
机构
[1] Univ Cincinnati, Coll Pharm, Div Pharmaceut Sci, Cincinnati, OH 45267 USA
[2] Childrens Hosp Med Ctr, Div Crit Care, Cincinnati, OH USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY | 2005年 / 288卷 / 03期
关键词
Caco-2; cells; cytokines; efflux systems; signal transduction pathways;
D O I
10.1152/ajpgi.00248.2004
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Patients with refractory inflammatory bowel disease (IBD) exhibit increased expression of intestinal P-glycoprotein (P-gp) as well as elevated luminal IFN-gamma and nitric oxide ( NO) levels. Using the in vitro Caco-2 cell culture model, we investigated whether these pathological mediators associated with the etiology of IBD affect functional activity of intestinal efflux systems. IFN-gamma reduced cellular uptake of cyclosporin A (CysA) but not methotrexate (MTX) in a time- and concentration-dependent manner. Simultaneously, P-gp expression increased by approximately twofold. Coincubation with the inducible NO synthase inhibitor L-N-6-(1-iminoethyl)-lysine (L-NIL) dramatically reduced production of intracellular NO in response to IFN-gamma stimulus. The presence of L-NIL also abrogated the cytokine-mediated increase in P-gp expression and function suggesting that NO is required for IFN-gamma-mediated activation of this efflux system. Exposure of Caco-2 cells to the chemical NO donor S-nitroso-N-acetylpenicillamine ( SNAP) produced a concentration-dependent decrease in intracellular CysA accumulation that was paralleled by an increase in P-gp expression. Both IFN-gamma and SNAP enhanced DNA binding of NF-kappaB, whereas inclusion of L-NIL dramatically decreased this cytokine-induced effect on NF-kappaB binding. These results suggest that NO mediates IFN-gamma-induced increase in expression and function of intestinal P-gp in the human Caco-2 cell culture model by altering DNA binding of NF-kappaB, which may enhance transcription of the ABCB1 gene encoding for this efflux system.
引用
收藏
页码:G533 / G540
页数:8
相关论文
共 49 条
[1]   P-glycoprotein: from genomics to mechanism [J].
Ambudkar, SV ;
Kimchi-Sarfaty, C ;
Sauna, ZE ;
Gottesman, MM .
ONCOGENE, 2003, 22 (47) :7468-7485
[2]   Messenger RNA expression of transporter and ion channel genes in undifferentiated and differentiated Caco-2 cells compared to human intestines [J].
Anderle, P ;
Rakhmanova, V ;
Woodford, K ;
Zerangue, N ;
Sadée, W .
PHARMACEUTICAL RESEARCH, 2003, 20 (01) :3-15
[3]   Effect of tumor necrosis factor-α and interferon-γ on intestinal P-glycoprotein expression, activity, and localization in Caco-2 cells [J].
Belliard, AM ;
Lacour, B ;
Farinotti, R ;
Leroy, C .
JOURNAL OF PHARMACEUTICAL SCIENCES, 2004, 93 (06) :1524-1536
[4]   Transporter-enzyme interactions:: Implications for predicting drug-drug interactions from in vitro data [J].
Benet, LZ ;
Cummins, CL ;
Wu, CY .
CURRENT DRUG METABOLISM, 2003, 4 (05) :393-398
[5]   NF-κB transcription factor induces drug resistance through MDR1 expression in cancer cells [J].
Bentires-Alj, M ;
Barbu, V ;
Fillet, M ;
Chariot, A ;
Relic, B ;
Jacobs, N ;
Gielen, J ;
Merville, MP ;
Bours, V .
ONCOGENE, 2003, 22 (01) :90-97
[6]  
Bertilsson PM, 2001, J PHARM SCI, V90, P638, DOI 10.1002/1520-6017(200105)90:5<638::AID-JPS1020>3.0.CO
[7]  
2-L
[8]   The immunological and genetic basis of inflammatory bowel disease [J].
Bouma, G ;
Strober, W .
NATURE REVIEWS IMMUNOLOGY, 2003, 3 (07) :521-533
[9]   Regulation of induction of nitric oxide synthase and the inhibitory actions of dexamethasone in the human intestinal epithelial cell line, Caco-2: influence of cell differentiation [J].
Cavicchi, M ;
Whittle, BJR .
BRITISH JOURNAL OF PHARMACOLOGY, 1999, 128 (03) :705-715
[10]   Acquired interferonγ responsiveness during Caco-2 cell differentiation:: effects on iNOS gene expression [J].
Chavez, AM ;
Morin, MJ ;
Unno, N ;
Fink, MP ;
Hodin, RA .
GUT, 1999, 44 (05) :659-665