Tissue-nonspecific Alkaline Phosphatase Is Activated in Enterocytes by Oxidative Stress Via Changes in Glycosylation

被引:60
作者
Lopez-Posadas, Rocio [1 ]
Gonzalez, Raquel [1 ]
Ballester, Isabel [2 ]
Martinez-Moya, Patricia [3 ]
Romero-Calvo, Isabel [1 ]
Dolores Suarez, Maria [3 ]
Zarzuelo, Antonio [1 ]
Martinez-Augustin, Olga [3 ]
Sanchez de Medina, Fermin [1 ]
机构
[1] Univ Granada, Sch Pharm, Dept Pharmacol, Ctr Invest Biomed Red Enfermedades Hepat & Digest, E-18071 Granada, Spain
[2] Massachusetts Gen Hosp, Gastrointestinal Unit GRJ716, Boston, MA 02114 USA
[3] Univ Granada, Sch Pharm, Dept Biochem & Mol Biol, Ctr Invest Biomed Red Enfermedades Hepat & Digest, E-18071 Granada, Spain
关键词
alkaline phosphatase; oxidative stress; inflammatory bowel disease; glycosylation; EPITHELIAL-CELL LINE; INDUCED RAT COLITIS; MESSENGER-RNA; SINGLE-GENE; IN-VIVO; LIVER; KIDNEY; BONE; EXPRESSION; SERUM;
D O I
10.1002/ibd.21381
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Background: Intestinal inflammation produces an induction of alkaline phosphatase (AP) activity that is attributable in part to augmented expression, accompanied by a change in isoform, in epithelial cells. Methods: This study focuses on induction of AP in intestinal epithelial cells in vitro. Results: Treatment with the oxidants H2O2, monochloramine, or tButOOH increases AP activity in vitro in Caco-2, HT29, and IEC18 cells. We selected IEC18 cells for further testing. Basal AP activity in IEC18 cells is of the tissue-nonspecific (bone-liver-kidney) type, as indicated by Northern and Western blot analysis. Oxidative stress augments AP activity and the sensitivity of the enzyme to levamisole, homoarginine, and heat in IEC18 cells. Increased immunoreactivity to tissue-nonspecific AP antibodies suggests an isoform shift from liver to either kidney or bone type. This effect occurs without changes at the mRNA level and is sensitive to tunicamycin, an inhibitor of N-glycosylation, and neuraminidase digestion. Saponin and deoxycholate produce similar effects to oxidants. Butyrate but not proinflammatory cytokines or LPS can induce a similar effect but without toxicity. The AP increase is not prevented by modulators of the MAPK, NF-kappa B, calcium, and cyclic adenosine monophosphate (cAMP) pathways, and is actually enhanced by actinomycin D via higher cell stress. Conclusions: Oxidative stress causes a distinct increase in enterocyte AP activity together with cell toxicity via changes in the glycosylation of the enzyme that correspond to a shift in isotype within the tissue-nonspecific paradigm. We speculate that this may have physiological implication for gut defense.
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页码:543 / 556
页数:14
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