Chymase inhibitor TY-51469 in therapy of inflammatory bowel disease
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摘要
AIM: To investigate the effect of chymase inhibitor TY-51469 in the therapy of inflammatory bowel disease and the underlying mechanism. METHODS: Seventy-five healthy Sprague-Dawley rats were randomly assigned to one of the three groups(control group, model group and TY-51469 experiment group) and each group had twenty-five rats. The rats of the model group and experiment group were subjected to treatment with 3.5% dextran sulfate sodium(DSS) 10 mg/kg to induce colitis. The control group and model group were subjected to intraperitoneal injection of saline, while the experiment group was subjected to intraperitoneal injection of 10 mg/kg TY-51469 each day. Five rats of each group were sacrificed on 0, 7, 14, 21 and 28 d, respectively. The degree of inflammation was assessed by histopathological scoring; flow cytometry was performed to detect the proportion of CD4+CD25+ Tregs in peripheral blood; colon tissues of rats were collected to measure m RNA and protein expression by PCR, Western blot and immunohistochemistry; serum levels of interleukin(IL)-10, transforming growth factor(TGF)-β1 and IL-17A were detected by ELISA. RESULTS: The rats in the experiment group and model group had significantly more severe colitis than the ones in the control group(P < 0.05) before treatment on day 0; no significant difference was observed between the experiment group and model group(P > 0.05). After treatment with TY-51469, the rats in the experiment group had significantly less severe colitis compared with the model group on 7, 14, 21 and 28 d(P < 0.05). The proportion of CD4+CD25+ Tregs was lower in the model group and experiment group than in the control group; the experiment group had a significantly higher proportion of CD4+CD25+ Tregs than that in the model group(P < 0.05). The model group and experiment group demonstrated lower expression of Foxp3 than the control group; the experiment group had higher Foxp3 expression than the model group(P < 0.05). Cytokines IL-10, TGF-β1 and IL-17 A were lower in the model group and experiment group than in the control group; the experiment group had higher expression than the model group(P < 0.05). CONCLUSION: After treatment with chymase inhibitor TY-51469, the experiment group demonstrated more significantly reduced intestinal inflammation and higher expression of immune tolerance related cytokines(IL-10, TGF-β1, IL-17A) and Foxp3 which is specifically expressed in Tregs compared with the model group. Therefore, chymase inhibitor TY-51469 might ameliorate the progression of DSS-induced colitis possibly by increasing the expression of Tregs and cytokines.
AIM: To investigate the effect of chymase inhibitor TY-51469 in the therapy of inflammatory bowel disease and the underlying mechanism. METHODS: Seventy-five healthy Sprague-Dawley rats were randomly assigned to one of the three groups(control group, model group and TY-51469 experiment group) and each group had twenty-five rats. The rats of the model group and experiment group were subjected to treatment with 3.5% dextran sulfate sodium(DSS) 10 mg/kg to induce colitis. The control group and model group were subjected to intraperitoneal injection of saline, while the experiment group was subjected to intraperitoneal injection of 10 mg/kg TY-51469 each day. Five rats of each group were sacrificed on 0, 7, 14, 21 and 28 d, respectively. The degree of inflammation was assessed by histopathological scoring; flow cytometry was performed to detect the proportion of CD4+CD25+ Tregs in peripheral blood; colon tissues of rats were collected to measure m RNA and protein expression by PCR, Western blot and immunohistochemistry; serum levels of interleukin(IL)-10, transforming growth factor(TGF)-β1 and IL-17A were detected by ELISA. RESULTS: The rats in the experiment group and model group had significantly more severe colitis than the ones in the control group(P < 0.05) before treatment on day 0; no significant difference was observed between the experiment group and model group(P > 0.05). After treatment with TY-51469, the rats in the experiment group had significantly less severe colitis compared with the model group on 7, 14, 21 and 28 d(P < 0.05). The proportion of CD4+CD25+ Tregs was lower in the model group and experiment group than in the control group; the experiment group had a significantly higher proportion of CD4+CD25+ Tregs than that in the model group(P < 0.05). The model group and experiment group demonstrated lower expression of Foxp3 than the control group; the experiment group had higher Foxp3 expression than the model group(P < 0.05). Cytokines IL-10, TGF-β1 and IL-17 A were lower in the model group and experiment group than in the control group; the experiment group had higher expression than the model group(P < 0.05). CONCLUSION: After treatment with chymase inhibitor TY-51469, the experiment group demonstrated more significantly reduced intestinal inflammation and higher expression of immune tolerance related cytokines(IL-10, TGF-β1, IL-17A) and Foxp3 which is specifically expressed in Tregs compared with the model group. Therefore, chymase inhibitor TY-51469 might ameliorate the progression of DSS-induced colitis possibly by increasing the expression of Tregs and cytokines.
AIM: To investigate the effect of chymase inhibitor TY-51469 in the therapy of inflammatory bowel disease and the underlying mechanism. METHODS: Seventy-five healthy Sprague-Dawley rats were randomly assigned to one of the three groups(control group, model group and TY-51469 experiment group) and each group had twenty-five rats. The rats of the model group and experiment group were subjected to treatment with 3.5% dextran sulfate sodium(DSS) 10 mg/kg to induce colitis. The control group and model group were subjected to intraperitoneal injection of saline, while the experiment group was subjected to intraperitoneal injection of 10 mg/kg TY-51469 each day. Five rats of each group were sacrificed on 0, 7, 14, 21 and 28 d, respectively. The degree of inflammation was assessed by histopathological scoring; flow cytometry was performed to detect the proportion of CD4+CD25+ Tregs in peripheral blood; colon tissues of rats were collected to measure m RNA and protein expression by PCR, Western blot and immunohistochemistry; serum levels of interleukin(IL)-10, transforming growth factor(TGF)-β1 and IL-17A were detected by ELISA. RESULTS: The rats in the experiment group and model group had significantly more severe colitis than the ones in the control group(P < 0.05) before treatment on day 0; no significant difference was observed between the experiment group and model group(P > 0.05). After treatment with TY-51469, the rats in the experiment group had significantly less severe colitis compared with the model group on 7, 14, 21 and 28 d(P < 0.05). The proportion of CD4+CD25+ Tregs was lower in the model group and experiment group than in the control group; the experiment group had a significantly higher proportion of CD4+CD25+ Tregs than that in the model group(P < 0.05). The model group and experiment group demonstrated lower expression of Foxp3 than the control group; the experiment group had higher Foxp3 expression than the model group(P < 0.05). Cytokines IL-10, TGF-β1 and IL-17 A were lower in the model group and experiment group than in the control group; the experiment group had higher expression than the model group(P < 0.05). CONCLUSION: After treatment with chymase inhibitor TY-51469, the experiment group demonstrated more significantly reduced intestinal inflammation and higher expression of immune tolerance related cytokines(IL-10, TGF-β1, IL-17A) and Foxp3 which is specifically expressed in Tregs compared with the model group. Therefore, chymase inhibitor TY-51469 might ameliorate the progression of DSS-induced colitis possibly by increasing the expression of Tregs and cytokines.
引文
1 Ford AC,Moayyedi P,Hanauer SB.Ulcerative colitis.BMJ 2013;346 :f432[PMID:23386404 DOI:10.1136/bmj.f432]
2 Di Sabatino A,Biancheri P,Rovedatti L,Macdonald TT,Corazza GR.Recent advances in understanding ulcerative colitis.Intern Emerg Med 2012;7:103-111[PMID:22068230 DOI:10.1007/s11739-011-0719-z]
3 Nielsen OH.New strategies for treatment of inflammatory bowel disease.Front Med(Lausanne)2014;1:3[PMID:25685754 DOI:10 .3389/fmed.2014.00003]
4 Amiot A,Peyrin-Biroulet L.Current,new and future biological agents on the horizon for the treatment of inflammatory bowel diseases.Therap Adv Gastroenterol 2015;8:66-82[PMID:25729432 DOI:10.1177/1756283X14558193]
5 Ananthakrishnan AN.Epidemiology and risk factors for IBD.Nat Rev Gastroenterol Hepatol 2015;12:205-217[PMID:25732745 DOI:10.1038/nrgastro.2015.34]
6 Maul J,Zeitz M.Ulcerative colitis:immune function,tissue fibrosis and current therapeutic considerations.Langenbecks Arch Surg 2012;397:1-10[PMID:21479621 DOI:10.1007/s00423-011-0789-4]
7 Dasgupta A,Saxena R.Regulatory T cells:a review.Natl Med J India 2012;25:341-351[PMID:23998865]
8 Oh SA,Li MO.TGF-β:guardian of T cell function.J Immunol 2013;191:3973-3979[PMID:24098055 DOI:10.4049/jimmunol.1301843]
9 Yahiro E,Miura S,Imaizumi S,Uehara Y,Saku K.Chymase inhibitors.Curr Pharm Des 2013;19:3065-3071[PMID:23176221]
10 Tojo H,Urata H.Chymase inhibition and cardiovascular protection.Cardiovasc Drugs Ther 2013;27:139-143[PMID:23468322 DOI:10.1007/s10557-013-6450-4]
11 Dutra RC,Cola M,Leite DF,Bento AF,Claudino RF,Nascimento AF,Leal PC,Calixto JB.Inhibitor of PI3Kγameliorates TNBSinduced colitis in mice by affecting the functional activity of CD4+CD25+Fox P3+regulatory T cells.Br J Pharmacol 2011;163:358-374[PMID:21244371 DOI:10.1111/j.1476-5381.2011.01226.x]
12 Xavier RJ,Podolsky DK.Unravelling the pathogenesis of inflammatory bowel disease.Nature 2007;448:427-434[PMID:17653185]
13 Cooney R,Jewell D.The genetic basis of inflammatory bowel disease.Dig Dis 2009;27:428-442[PMID:19897957 DOI:10 .1159/000234909]
14 Reiff C,Kelly D.Inflammatory bowel disease,gut bacteria and probiotic therapy.Int J Med Microbiol 2010;300:25-33[PMID:19800289 DOI:10.1016/j.ijmm.2009.08.004]
15 Jani N,Regueiro MD.Medical therapy for ulcerative colitis.Gastroenterol Clin North Am 2002;31:147-166[PMID:12122729]
16 Lakatos PL,Lakatos L.Ulcerative proctitis:a review of pharmacotherapy and management.Expert Opin Pharmacother 2008;9:741-749[PMID:18345952 DOI:10.1517/14656566.9.5.741]
17 Heuston S,Hyland NP.Chymase inhibition as a pharmacological target:a role in inflammatory and functional gastrointestinal disorders?Br J Pharmacol 2012;167:732-740[PMID:22646261DOI:10.1111/j.1476-5381.2012.02055.x]
18 Doggrell SA,Wanstall JC.Cardiac chymase:pathophysiological role and therapeutic potential of chymase inhibitors.Can J Physiol Pharmacol 2005;83:123-130[PMID:15791285]
19 Ishida K,Takai S,Murano M,Nishikawa T,Inoue T,Murano N,Inoue N,Jin D,Umegaki E,Higuchi K,Miyazaki M.Role of chymase-dependent matrix metalloproteinase-9 activation in mice with dextran sodium sulfate-induced colitis.J Pharmacol Exp Ther2008;324:422-426[PMID:18024785]
20 Kakimoto K,Takai S,Murano M,Ishida K,Yoda Y,Inoue T,Jin D,Umegaki E,Higuchi K.Significance of chymase-dependent matrix metalloproteinase-9 activation on indomethacin-induced small intestinal damages in rats.J Pharmacol Exp Ther 2010;332:684-689[PMID:19996300 DOI:10.1124/jpet.109.162933]
21 Bamias G,Kaltsa G,Ladas SD.Cytokines in the pathogenesis of ulcerative colitis.Discov Med 2011;11:459-467[PMID:21616044]
22 Mayne CG,Williams CB.Induced and natural regulatory T cells in the development of inflammatory bowel disease.Inflamm Bowel Dis 2013;19:1772-1788[PMID:23656897 DOI:10.1097/MIB.0b013e318281f5a3]
23 Lo WC,Arsenescu RI,Friedman A.Mathematical model of the roles of T cells in inflammatory bowel disease.Bull Math Biol 2013;75:1417-1433[PMID:23760658 DOI:10.1007/s11538-013-9853-2]
24 Marlow GJ,van Gent D,Ferguson LR.Why interleukin-10supplementation does not work in Crohn’s disease patients.World J Gastroenterol 2013;19:3931-3941[PMID:23840137 DOI:10 .3748/wjg.v19.i25.3931]
25 Han G,Li F,Singh TP,Wolf P,Wang XJ.The pro-inflammatory role of TGFβ1:a paradox?Int J Biol Sci 2012;8:228-235[PMID:22253566]
26 Buckner JH.Mechanisms of impaired regulation by CD4(+)CD25(+)FOXP3(+)regulatory T cells in human autoimmune diseases.Nat Rev Immunol 2010;10:849-859[PMID:21107346 DOI:10 .1038/nri2889]
27 Cao AT,Yao S,Gong B,Elson CO,Cong Y.Th17 cells upregulate polymeric Ig receptor and intestinal Ig A and contribute to intestinal homeostasis.J Immunol 2012;189:4666-4673[PMID:22993206DOI:10.4049/jimmunol.1200955]
28 Ogawa A,Andoh A,Araki Y,Bamba T,Fujiyama Y.Neutralization of interleukin-17 aggravates dextran sulfate sodium-induced colitis in mice.Clin Immunol 2004;110:55-62[PMID:14962796]
29 Gladiator A,Wangler N,Trautwein-Weidner K,Leibund GutLandmann S.Cutting edge:IL-17-secreting innate lymphoid cells are essential for host defense against fungal infection.J Immunol 2013;190:521-525[PMID:23255360 DOI:10.4049/jimmunol.1202924]
30 Puel A,Cypowyj S,Bustamante J,Wright JF,Liu L,Lim HK,Migaud M,Israel L,Chrabieh M,Audry M,Gumbleton M,Toulon A,Bodemer C,El-Baghdadi J,Whitters M,Paradis T,Brooks J,Collins M,Wolfman NM,Al-Muhsen S,Galicchio M,Abel L,Picard C,Casanova JL.Chronic mucocutaneous candidiasis in humans with inborn errors of interleukin-17 immunity.Science 2011;332:65-68[PMID:21350122 DOI:10.1126/science.1200439]
2 Di Sabatino A,Biancheri P,Rovedatti L,Macdonald TT,Corazza GR.Recent advances in understanding ulcerative colitis.Intern Emerg Med 2012;7:103-111[PMID:22068230 DOI:10.1007/s11739-011-0719-z]
3 Nielsen OH.New strategies for treatment of inflammatory bowel disease.Front Med(Lausanne)2014;1:3[PMID:25685754 DOI:10 .3389/fmed.2014.00003]
4 Amiot A,Peyrin-Biroulet L.Current,new and future biological agents on the horizon for the treatment of inflammatory bowel diseases.Therap Adv Gastroenterol 2015;8:66-82[PMID:25729432 DOI:10.1177/1756283X14558193]
5 Ananthakrishnan AN.Epidemiology and risk factors for IBD.Nat Rev Gastroenterol Hepatol 2015;12:205-217[PMID:25732745 DOI:10.1038/nrgastro.2015.34]
6 Maul J,Zeitz M.Ulcerative colitis:immune function,tissue fibrosis and current therapeutic considerations.Langenbecks Arch Surg 2012;397:1-10[PMID:21479621 DOI:10.1007/s00423-011-0789-4]
7 Dasgupta A,Saxena R.Regulatory T cells:a review.Natl Med J India 2012;25:341-351[PMID:23998865]
8 Oh SA,Li MO.TGF-β:guardian of T cell function.J Immunol 2013;191:3973-3979[PMID:24098055 DOI:10.4049/jimmunol.1301843]
9 Yahiro E,Miura S,Imaizumi S,Uehara Y,Saku K.Chymase inhibitors.Curr Pharm Des 2013;19:3065-3071[PMID:23176221]
10 Tojo H,Urata H.Chymase inhibition and cardiovascular protection.Cardiovasc Drugs Ther 2013;27:139-143[PMID:23468322 DOI:10.1007/s10557-013-6450-4]
11 Dutra RC,Cola M,Leite DF,Bento AF,Claudino RF,Nascimento AF,Leal PC,Calixto JB.Inhibitor of PI3Kγameliorates TNBSinduced colitis in mice by affecting the functional activity of CD4+CD25+Fox P3+regulatory T cells.Br J Pharmacol 2011;163:358-374[PMID:21244371 DOI:10.1111/j.1476-5381.2011.01226.x]
12 Xavier RJ,Podolsky DK.Unravelling the pathogenesis of inflammatory bowel disease.Nature 2007;448:427-434[PMID:17653185]
13 Cooney R,Jewell D.The genetic basis of inflammatory bowel disease.Dig Dis 2009;27:428-442[PMID:19897957 DOI:10 .1159/000234909]
14 Reiff C,Kelly D.Inflammatory bowel disease,gut bacteria and probiotic therapy.Int J Med Microbiol 2010;300:25-33[PMID:19800289 DOI:10.1016/j.ijmm.2009.08.004]
15 Jani N,Regueiro MD.Medical therapy for ulcerative colitis.Gastroenterol Clin North Am 2002;31:147-166[PMID:12122729]
16 Lakatos PL,Lakatos L.Ulcerative proctitis:a review of pharmacotherapy and management.Expert Opin Pharmacother 2008;9:741-749[PMID:18345952 DOI:10.1517/14656566.9.5.741]
17 Heuston S,Hyland NP.Chymase inhibition as a pharmacological target:a role in inflammatory and functional gastrointestinal disorders?Br J Pharmacol 2012;167:732-740[PMID:22646261DOI:10.1111/j.1476-5381.2012.02055.x]
18 Doggrell SA,Wanstall JC.Cardiac chymase:pathophysiological role and therapeutic potential of chymase inhibitors.Can J Physiol Pharmacol 2005;83:123-130[PMID:15791285]
19 Ishida K,Takai S,Murano M,Nishikawa T,Inoue T,Murano N,Inoue N,Jin D,Umegaki E,Higuchi K,Miyazaki M.Role of chymase-dependent matrix metalloproteinase-9 activation in mice with dextran sodium sulfate-induced colitis.J Pharmacol Exp Ther2008;324:422-426[PMID:18024785]
20 Kakimoto K,Takai S,Murano M,Ishida K,Yoda Y,Inoue T,Jin D,Umegaki E,Higuchi K.Significance of chymase-dependent matrix metalloproteinase-9 activation on indomethacin-induced small intestinal damages in rats.J Pharmacol Exp Ther 2010;332:684-689[PMID:19996300 DOI:10.1124/jpet.109.162933]
21 Bamias G,Kaltsa G,Ladas SD.Cytokines in the pathogenesis of ulcerative colitis.Discov Med 2011;11:459-467[PMID:21616044]
22 Mayne CG,Williams CB.Induced and natural regulatory T cells in the development of inflammatory bowel disease.Inflamm Bowel Dis 2013;19:1772-1788[PMID:23656897 DOI:10.1097/MIB.0b013e318281f5a3]
23 Lo WC,Arsenescu RI,Friedman A.Mathematical model of the roles of T cells in inflammatory bowel disease.Bull Math Biol 2013;75:1417-1433[PMID:23760658 DOI:10.1007/s11538-013-9853-2]
24 Marlow GJ,van Gent D,Ferguson LR.Why interleukin-10supplementation does not work in Crohn’s disease patients.World J Gastroenterol 2013;19:3931-3941[PMID:23840137 DOI:10 .3748/wjg.v19.i25.3931]
25 Han G,Li F,Singh TP,Wolf P,Wang XJ.The pro-inflammatory role of TGFβ1:a paradox?Int J Biol Sci 2012;8:228-235[PMID:22253566]
26 Buckner JH.Mechanisms of impaired regulation by CD4(+)CD25(+)FOXP3(+)regulatory T cells in human autoimmune diseases.Nat Rev Immunol 2010;10:849-859[PMID:21107346 DOI:10 .1038/nri2889]
27 Cao AT,Yao S,Gong B,Elson CO,Cong Y.Th17 cells upregulate polymeric Ig receptor and intestinal Ig A and contribute to intestinal homeostasis.J Immunol 2012;189:4666-4673[PMID:22993206DOI:10.4049/jimmunol.1200955]
28 Ogawa A,Andoh A,Araki Y,Bamba T,Fujiyama Y.Neutralization of interleukin-17 aggravates dextran sulfate sodium-induced colitis in mice.Clin Immunol 2004;110:55-62[PMID:14962796]
29 Gladiator A,Wangler N,Trautwein-Weidner K,Leibund GutLandmann S.Cutting edge:IL-17-secreting innate lymphoid cells are essential for host defense against fungal infection.J Immunol 2013;190:521-525[PMID:23255360 DOI:10.4049/jimmunol.1202924]
30 Puel A,Cypowyj S,Bustamante J,Wright JF,Liu L,Lim HK,Migaud M,Israel L,Chrabieh M,Audry M,Gumbleton M,Toulon A,Bodemer C,El-Baghdadi J,Whitters M,Paradis T,Brooks J,Collins M,Wolfman NM,Al-Muhsen S,Galicchio M,Abel L,Picard C,Casanova JL.Chronic mucocutaneous candidiasis in humans with inborn errors of interleukin-17 immunity.Science 2011;332:65-68[PMID:21350122 DOI:10.1126/science.1200439]