摘要
AIM: To investigate the effects of nilotinib in a rat model of indomethacin-induced enterocolitis.METHODS: Twenty-one Wistar albino female rats obtained from Dokuz Eylul University Department of Laboratory Animal Science were divided into the following three groups: control(n = 7), indomethacin(n = 7) and nilotinib(n = 7). A volume of 0.25 m L of physiological serum placebo was administered to the control and indomethacin groups through an orogastric tube for 13 d. To induce enterocolitis, the indomethacin and nilotinib groups received 7.5 m L/kg indomethacin dissolved in 5% sodium bicarbonate and administered subcutaneously in a volume of 0.5 m L twice daily for three days. Nilotinib was administered 20 mg/kg/d in two divided doses to the nilotinib group of rats for 13 d through an orogastric tube, beginning on the same day as indomethacin administration. For 13 d, the rats werefed a standard diet, and their weights were monitored daily. After the rats were sacrificed, the intestinal and colonic tissue samples were examined. The macroscopic and microscopic pathology scores were evaluated. The pathologist stained all tissue samples using terminal deoxynucleotidyl transferase-mediated d UTP-biotin nick-end labeling method. Mucosal crypts and apoptotic cells were quantified. The plateletderived growth factor receptor(PDGFR) α and β scores assessed by immunohistochemical staining method and tissue and serum tumor necrosis factor(TNF) α levels were determined by enzyme-linked immunosorbent assay. RESULTS: Between days 1 and 13, the rats in the nilotinib and indomethacin groups lost significantly more weight than the controls(-11 g vs +14.14 g, P = 0.013;-30 g vs +14.14 g, P = 0.003). In the small intestinal and colonic tissues, the macroscopic scores were significantly lower in the nilotinib group than in the indomethacin group(1.14 ± 0.38 and 7.29 ± 2.98, P = 0.005; 1.14 ± 0.38 and 7.43 ± 2.64, P = 0.001, respectively), but the values of the nilotinib and indomethacin groups were similar to the control group. In the small intestinal and colonic tissues, the microscopic scores were significantly lower in the nilotinib group than in the indomethacin group(3.43 ± 2.99 and 7.67 ± 3.67, P = 0.043; 2.29 ± 0.76 and 8.80 ± 2.68, P = 0.003, respectively), but the values were similar to the control group. The PDGFR β scores in the small intestine and colon were significantly lower in the nilotinib group than in the indomethacin group(1.43 ± 0.79 and 2.43 ± 0.54, P = 0.021; 1.57 ± 0.54 and 3 ± 0, P =0.001), and the values were similar to controls. The colonic PDGFR α scores were significantly lower in the nilotinib group than in the indomethacin group(1.71 ± 0.49 and 3 ± 0, P = 0.001). The colonic apoptosis scores were significantly lower in the controls than in the nilotinib group(1.57 ± 1.13 and 4 ± 1.29, P = 0.007). Furthermore, the serum and tissue TNF-α levels were similar between the nilotinib and indomethacin groups.C O N C L U S I O N : In the indomethacin-induced enterocolitis rat model, nilotinib has a positive effect on the macroscopic and microscopic pathologic scores, ensuring considerable mucosal healing. Nilotinib decreases PDGFR α and β levels and increases the colonic apoptotic scores, but it has no significant effects on weight loss and the TNF-α levels.
AIM: To investigate the effects of nilotinib in a rat model of indomethacin-induced enterocolitis.METHODS: Twenty-one Wistar albino female rats obtained from Dokuz Eylul University Department of Laboratory Animal Science were divided into the following three groups: control(n = 7), indomethacin(n = 7) and nilotinib(n = 7). A volume of 0.25 m L of physiological serum placebo was administered to the control and indomethacin groups through an orogastric tube for 13 d. To induce enterocolitis, the indomethacin and nilotinib groups received 7.5 m L/kg indomethacin dissolved in 5% sodium bicarbonate and administered subcutaneously in a volume of 0.5 m L twice daily for three days. Nilotinib was administered 20 mg/kg/d in two divided doses to the nilotinib group of rats for 13 d through an orogastric tube, beginning on the same day as indomethacin administration. For 13 d, the rats werefed a standard diet, and their weights were monitored daily. After the rats were sacrificed, the intestinal and colonic tissue samples were examined. The macroscopic and microscopic pathology scores were evaluated. The pathologist stained all tissue samples using terminal deoxynucleotidyl transferase-mediated d UTP-biotin nick-end labeling method. Mucosal crypts and apoptotic cells were quantified. The plateletderived growth factor receptor(PDGFR) α and β scores assessed by immunohistochemical staining method and tissue and serum tumor necrosis factor(TNF) α levels were determined by enzyme-linked immunosorbent assay. RESULTS: Between days 1 and 13, the rats in the nilotinib and indomethacin groups lost significantly more weight than the controls(-11 g vs +14.14 g, P = 0.013;-30 g vs +14.14 g, P = 0.003). In the small intestinal and colonic tissues, the macroscopic scores were significantly lower in the nilotinib group than in the indomethacin group(1.14 ± 0.38 and 7.29 ± 2.98, P = 0.005; 1.14 ± 0.38 and 7.43 ± 2.64, P = 0.001, respectively), but the values of the nilotinib and indomethacin groups were similar to the control group. In the small intestinal and colonic tissues, the microscopic scores were significantly lower in the nilotinib group than in the indomethacin group(3.43 ± 2.99 and 7.67 ± 3.67, P = 0.043; 2.29 ± 0.76 and 8.80 ± 2.68, P = 0.003, respectively), but the values were similar to the control group. The PDGFR β scores in the small intestine and colon were significantly lower in the nilotinib group than in the indomethacin group(1.43 ± 0.79 and 2.43 ± 0.54, P = 0.021; 1.57 ± 0.54 and 3 ± 0, P =0.001), and the values were similar to controls. The colonic PDGFR α scores were significantly lower in the nilotinib group than in the indomethacin group(1.71 ± 0.49 and 3 ± 0, P = 0.001). The colonic apoptosis scores were significantly lower in the controls than in the nilotinib group(1.57 ± 1.13 and 4 ± 1.29, P = 0.007). Furthermore, the serum and tissue TNF-α levels were similar between the nilotinib and indomethacin groups.C O N C L U S I O N : In the indomethacin-induced enterocolitis rat model, nilotinib has a positive effect on the macroscopic and microscopic pathologic scores, ensuring considerable mucosal healing. Nilotinib decreases PDGFR α and β levels and increases the colonic apoptotic scores, but it has no significant effects on weight loss and the TNF-α levels.
引文
1 Loftus EV.Clinical epidemiology of inflammatory bowel disease:Incidence,prevalence,and environmental influences.Gastroenterology 2004;126:1504-1517[PMID:15168363 DOI:10 .1053/j.gastro.2004.01.063]
2 Hanauer SB,Present DH.The state of the art in the management of inflammatory bowel disease.Rev Gastroenterol Disord 2003;3:81-92[PMID:12776005]
3 Sandborn WJ,Feagan BG.Review article:mild to moderate Crohn’s disease--defining the basis for a new treatment algorithm.Aliment Pharmacol Ther 2003;18:263-277[PMID:12895211DOI:10.1046/j.1365-2036.2003.01661.x]
4 CammàC,Giunta M,Rosselli M,Cottone M.Mesalamine in the maintenance treatment of Crohn’s disease:a meta-analysis adjusted for confounding variables.Gastroenterology 1997;113:1465-1473[PMID:9352848 DOI:10.1053/gast.1997.v113.pm9352848]
5 Lochs H,Mayer M,Fleig WE,Mortensen PB,Bauer P,Genser D,Petritsch W,Raithel M,Hoffmann R,Gross V,Plauth M,Staun M,Nesje LB.Prophylaxis of postoperative relapse in Crohn’s disease with mesalamine:European Cooperative Crohn’s Disease Study VI.Gastroenterology 2000;118:264-273[PMID:10648454 DOI:10 .1016/S0016-5085(00)70208-3]
6 Stein RB,Hanauer SB.Medical therapy for inflammatory bowel disease.Gastroenterol Clin North Am 1999;28:297-321[PMID:10372270 DOI:10.1016/S0889-8553(05)70058-3]
7 Feagan BG,Rochon J,Fedorak RN,Irvine EJ,Wild G,Sutherland L,Steinhart AH,Greenberg GR,Gillies R,Hopkins M.Methotrexate for the treatment of Crohn’s disease.The North American Crohn’s Study Group Investigators.N Engl J Med 1995;332:292-297[PMID:7816064 DOI:10.1056/NEJM199502023320503]
8 Feagan BG,Fedorak RN,Irvine EJ,Wild G,Sutherland L,Steinhart AH,Greenberg GR,Koval J,Wong CJ,Hopkins M,Hanauer SB,Mc Donald JW.A comparison of methotrexate with placebo for the maintenance of remission in Crohn’s disease.North American Crohn’s Study Group Investigators.N Engl J Med 2000;342:1627-1632[PMID:10833208 DOI:10.1056/NEJM200006013422202]
9 Williams I,Goh J.Investigational new drugs in the treatment of inflammatory bowel disease:a review.J Exp Pharmacol 2011;3:13-19[DOI:10.2147/JEP.S7806]
10 Ardizzone S,Bianchi Porro G.Inflammatory bowel disease:new insights into pathogenesis and treatment.J Intern Med2002;252:475-496[PMID:12472908 DOI:10.1046/j.1365-2796.2002.01067.x]
11 Kakkar A,Wasan SK,Farraye FA.Targeting mucosal healing in Crohn’s disease.Gastroenterol Hepatol(N Y)2011;7:374-380[PMID:21869869]
12 Landi B,Anh TN,Cortot A,Soule JC,Rene E,Gendre JP,Bories P,See A,Metman EH,Florent C.Endoscopic monitoring of Crohn’s disease treatment:a prospective,randomized clinical trial.The Groupe d’Etudes Therapeutiques des Affections Inflammatoires Digestives.Gastroenterology 1992;102:1647-1653[PMID:1568574]
13 Modigliani R,Mary JY,Simon JF,Cortot A,Soule JC,Gendre JP,Rene E.Clinical,biological,and endoscopic picture of attacks of Crohn’s disease.Evolution on prednisolone.Groupe d’Etude Thérapeutique des Affections Inflammatoires Digestives.Gastroenterology 1990;98:811-818[PMID:2179031]
14 Ma?osa M,Naves JE,Leal C,CabréE,Moreno V,LorenzoZu?iga V,Boix J,Domènech E.Does methotrexate induce mucosal healing in Crohn’s disease?Inflamm Bowel Dis 2010;16:377-378[PMID:19575354 DOI:10.1002/ibd.21015]
15 Laharie D,Reffet A,Belleannée G,Chabrun E,Subtil C,Razaire S,Capdepont M,de Lédinghen V.Mucosal healing with methotrexate in Crohn’s disease:a prospective comparative study with azathioprine and infliximab.Aliment Pharmacol Ther 2011;33:714-721[PMID:21235604 DOI:10.1111/j.1365-2036.2010.04569.x]
16 Sandborn WJ,Van O EC,Zins BJ,Tremaine WJ,Mays DC,Lipsky JJ.An intravenous loading dose of azathioprine decreases the time to response in patients with Crohn’s disease.Gastroenterology 1995;109:1808-1817[PMID:7498645 DOI:10 .1016/0016-5085(95)90747-5]
17 D’Haens G,Geboes K,Ponette E,Penninckx F,Rutgeerts P.Healing of severe recurrent ileitis with azathioprine therapy in patients with Crohn’s disease.Gastroenterology1997;112:1475-1481[PMID:9136824 DOI:10.1016/S0016-5085(97)70027-1]
18 D’Haens G,Geboes K,Rutgeerts P.Endoscopic and histologic healing of Crohn’s(ileo-)colitis with azathioprine.Gastrointest Endosc 1999;50:667-671[PMID:10536324]
19 D’Haens GR,Vermeire S,Van Assche G,Noman M,Aerden I,Van Olmen G,Rutgeerts P.Therapy of metronidazole with azathioprine to prevent postoperative recurrence of Crohn’s disease:a controlled randomized trial.Gastroenterology 2008;135:1123-1129[PMID:18727929 DOI:10.1053/j.gastro.2008.07.010]
20 Colombel JF,Sandborn WJ,Reinisch W,Mantzaris GJ,Kornbluth A,Rachmilewitz D,Lichtiger S,D’Haens G,Diamond RH,Broussard DL,Tang KL,van der Woude CJ,Rutgeerts P.Infliximab,azathioprine,or combination therapy for Crohn’s disease.N Engl J Med 2010;362:1383-1395[PMID:20393175DOI:10.1056/NEJMoa0904492]
21 Rutgeerts P,Feagan BG,Lichtenstein GR,Mayer LF,Schreiber S,Colombel JF,Rachmilewitz D,Wolf DC,Olson A,Bao W,Hanauer SB.Comparison of scheduled and episodic treatment strategies of infliximab in Crohn’s disease.Gastroenterology 2004;126:402-413[PMID:14762776 DOI:10.1053/j.gastro.2003.11.014]
22 Hanauer SB,Feagan BG,Lichtenstein GR,Mayer LF,Schreiber S,Colombel JF,Rachmilewitz D,Wolf DC,Olson A,Bao W,Rutgeerts P.Maintenance infliximab for Crohn’s disease:the ACCENT I randomised trial.Lancet 2002;359:1541-1549[PMID:12047962 DOI:10.1016/S0140-6736(02)08512-4]
23 Rutgeerts P,Van Assche G,Sandborn WJ,Wolf DC,Geboes K,Colombel JF,Reinisch W,Kumar A,Lazar A,Camez A,Lomax KG,Pollack PF,D’Haens G.Adalimumab induces and maintains mucosal healing in patients with Crohn’s disease:data from the EXTEND trial.Gastroenterology 2012;142:1102-1111.e2[PMID:22326435 DOI:10.1053/j.gastro.2012.01.035]
24 Colombel JF,Lémann M,Bouhnik Y.Endoscopic Mucosal Improvement in Patients With Active Crohn’s Disease Treated With Certolizumab Pegol:Week 10 and 54 Results of the Music Trial.Gastroenterology AGA 2010;138:S166[DOI:10.1016/S0016-5085(10)60761-5]
25 Sandborn WJ,Colombel JF,Enns R,Feagan BG,Hanauer SB,Lawrance IC,Panaccione R,Sanders M,Schreiber S,Targan S,van Deventer S,Goldblum R,Despain D,Hogge GS,Rutgeerts P.Natalizumab induction and maintenance therapy for Crohn’s disease.N Engl J Med 2005;353:1912-1925[PMID:16267322DOI:10.1056/NEJMoa043335]
26 Papi C,Fascì-Spurio F,Rogai F,Settesoldi A,Margagnoni G,Annese V.Mucosal healing in inflammatory bowel disease:treatment efficacy and predictive factors.Dig Liver Dis 2013;45:978-985[PMID:24018244 DOI:10.1016/j.dld.2013.07.006]
27 Dave M,Loftus EV.Mucosal healing in inflammatory bowel disease-a true paradigm of success?Gastroenterol Hepatol(N Y)2012;8:29-38[PMID:22347830]
28 Robinson DR,Wu YM,Lin SF.The protein tyrosine kinase family of the human genome.Oncogene 2000;19:5548-5557[PMID:11114734 DOI:10.1038/sj.onc.1203957]
29 Manley PW,Stiefl N,Cowan-Jacob SW,Kaufman S,Mestan J,Wartmann M,Wiesmann M,Woodman R,Gallagher N.Structural resemblances and comparisons of the relative pharmacological properties of imatinib and nilotinib.Bioorg Med Chem 2010;18:6977-6986[PMID:20817538 DOI:10.1016/j.bmc.2010.08.026]
30 Weisberg E,Manley PW,Breitenstein W,Brüggen J,CowanJacob SW,Ray A,Huntly B,Fabbro D,Fendrich G,Hall-Meyers E,Kung AL,Mestan J,Daley GQ,Callahan L,Catley L,Cavazza C,Azam M,Neuberg D,Wright RD,Gilliland DG,Griffin JD.Characterization of AMN107,a selective inhibitor of native and mutant Bcr-Abl.Cancer Cell 2005;7:129-141[PMID:15710326DOI:10.1016/j.ccr.2005.01.007]
31 Weisberg E,Manley P,Mestan J,Cowan-Jacob S,Ray A,Griffin JD.AMN107(nilotinib):a novel and selective inhibitor of BCRABL.Br J Cancer 2006;94:1765-1769[PMID:16721371 DOI:10 .1038/sj.bjc.6603170]
32 Manley PW,Drueckes P,Fendrich G,Furet P,Liebetanz J,Martiny-Baron G,Mestan J,Trappe J,Wartmann M,Fabbro D.Extended kinase profile and properties of the protein kinase inhibitor nilotinib.Biochim Biophys Acta 2010;1804:445-453[PMID:19922818 DOI:10.1016/j.bbapap.2009.11.008]
33 Rhee CK,Lee SH,Yoon HK,Kim SC,Lee SY,Kwon SS,Kim YK,Kim KH,Kim TJ,Kim JW.Effect of nilotinib on bleomycin-induced acute lung injury and pulmonary fibrosis in mice.Respiration 2011;82:273-287[PMID:21659722 DOI:10.1159/000327719]
34 Ataca P,Soyturk M,Karaman M,Unlu M,Sagol O,Dervis Hakim G,Yilmaz O.Nilotinib-mediated mucosal healing in a rat model of colitis.World J Gastroenterol 2013;19:6237-6244[PMID:24115822 DOI:10.3748/wjg.v19.i37.6237]
35 Yamada T,Deitch E,Specian RD,Perry MA,Sartor RB,Grisham MB.Mechanisms of acute and chronic intestinal inflammation induced by indomethacin.Inflammation 1993;17:641-662[PMID:7906675 DOI:10.1007/BF00920471]
36 Kawada M,Arihiro A,Mizoguchi E.Insights from advances in research of chemically induced experimental models of human inflammatory bowel disease.World J Gastroenterol 2007;13:5581-5593[PMID:17948932 DOI:10.3748/wjg.v13.i42.5581]
37 Vilaseca J,Salas A,Guarner F,Rodríguez R,Martínez M,Malagelada JR.Dietary fish oil reduces progression of chronic inflammatory lesions in a rat model of granulomatous colitis.Gut1990;31:539-544[PMID:2161781 DOI:10.1136/gut.31.5.539]
38 Dieleman LA,Palmen MJ,Akol H,Bloemena E,Pe?a AS,Meuwissen SG,Van Rees EP.Chronic experimental colitis induced by dextran sulphate sodium(DSS)is characterized by Th1 and Th2cytokines.Clin Exp Immunol 1998;114:385-391[PMID:9844047DOI:10.1046/j.1365-2249.1998.00728.x]
39 Baumgart DC,Carding SR.Inflammatory bowel disease:cause and immunobiology.Lancet 2007;369:1627-1640[PMID:17499605 DOI:10.1016/S0140-6736(07)60750-8]
40 Engel MA,Neurath MF.New pathophysiological insights and modern treatment of IBD.J Gastroenterol 2010;45:571-583[PMID:20213337 DOI:10.1007/s00535-010-0219-3]
41 Schlessinger J.Cell signaling by receptor tyrosine kinases.Cell 2000;103:211-225[PMID:11057895 DOI:10.1016/S0092-8674(00)00114-8]
42 Cuzzocrea S,Mc Donald MC,Mazzon E,Mota-Filipe H,Lepore V,Ciccolo A,Terranova ML,Britti D,Caputi AP,Thiemermann C.The tyrosine kinase inhibitor tyrphostin AG 126 reduced the development of colitis in the rat.Lab Invest 2000;80:1439-1453[PMID:11005212 DOI:10.1038/Labinvest.3780151]
43 Magro F,Costa C.Long-standing remission of Crohn’s disease under imatinib therapy in a patient with Crohn’s disease.Inflamm Bowel Dis 2006;12:1087-1089[PMID:17075351 DOI:10 .1097/01.mib.0000232468.15950.34]
44 Krzystek-Korpacka M,Neubauer K,Matusiewicz M.Plateletderived growth factor-BB reflects clinical,inflammatory and angiogenic disease activity and oxidative stress in inflammatory bowel disease.Clin Biochem 2009;42:1602-1609[PMID:19679116 DOI:10.1016/j.clinbiochem.2009.08.002]
45 Bonner JC.Regulation of PDGF and its receptors in fibrotic diseases.Cytokine Growth Factor Rev 2004;15:255-273[PMID:15207816 DOI:10.1016/j.cytogfr.2004.03.006]
46 Baumann H,Gauldie J.The acute phase response.Immunol Today1994;15:74-80[PMID:7512342 DOI:10.1016/0167-5699(94)90137-6]
47 Reimund JM,Wittersheim C,Dumont S,Muller CD,Baumann R,Poindron P,Duclos B.Mucosal inflammatory cytokine production by intestinal biopsies in patients with ulcerative colitis and Crohn’s disease.J Clin Immunol 1996;16:144-150[PMID:8734357 DOI:10 .1007/BF01540912]
48 De Backer O,Lefebvre RA.Is there a role for imatinib in inflammatory bowel disease?Inflamm Bowel Dis 2008;14:579-581[PMID:18050299 DOI:10.1002/ibd.20320]
49 Nair DG,Miller KG,Lourenssen SR,Blennerhassett MG.Inflammatory cytokines promote growth of intestinal smooth muscle cells by induced expression of PDGF-Rβ.J Cell Mol Med2014;18:444-454[PMID:24417820 DOI:10.1111/jcmm.12193]
50 Liu Y,Wang Z,Kwong SQ,Lui EL,Friedman SL,Li FR,Lam RW,Zhang GC,Zhang H,Ye T.Inhibition of PDGF,TGF-β,and Abl signaling and reduction of liver fibrosis by the small molecule Bcr-Abl tyrosine kinase antagonist Nilotinib.J Hepatol 2011;55:612-625[PMID:21251937 DOI:10.1016/j.jhep.2010.11.035]
51 Wang Y,Mandal D,Wang S,Kleinerman ES,Pollock RE,Lev D,Hayes-Jordan A.Platelet-derived growth factor receptor beta inhibition increases tumor necrosis factor-related apoptosisinducing ligand(TRAIL)sensitivity:imatinib and TRAIL dual therapy.Cancer 2010;116:3892-3902[PMID:20564078 DOI:10 .1002/cncr.25107]
52 D’Argenio G,Farrace MG,Cosenza V,De Ritis F,Della Valle N,Manguso F,Piacentini M.Expression of apoptosis-related proteins in rat with induced colitis.Int J Colorectal Dis 2004;19:451-460[PMID:15083321 DOI:10.1007/s00384-004-0585-5]