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Planta Med 2021; 87(01/02): 169-176
DOI: 10.1055/a-1202-4799
Biological and Pharmacological Activities
Original Papers

Bauhinia Protease Inhibitors Attenuate Gastric Ulcer by Blocking Neutrophil Enzymes

Mayara Vioto Valois
1   Department of Biochemistry, Universidade Federal de São Paulo, Escola Paulista de Medicina, SP, Brazil
,
Cleide de Oliveira
1   Department of Biochemistry, Universidade Federal de São Paulo, Escola Paulista de Medicina, SP, Brazil
,
Antonio José Lapa
2   Department of Pharmacology, Universidade Federal de São Paulo, Escola Paulista de Medicina, SP, Brazil
3   Universidade do Estado do Amazonas, Manaus, Amazonas, Brazil
,
Caden Souccar
2   Department of Pharmacology, Universidade Federal de São Paulo, Escola Paulista de Medicina, SP, Brazil
,
Maria Luiza Vilela Oliva 
1   Department of Biochemistry, Universidade Federal de São Paulo, Escola Paulista de Medicina, SP, Brazil
› Author AffiliationsSupported by: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Finance Code 001
Supported by: Fundação de Amparo à Pesquisa do Estado de São Paulo 72017/06630-2017/07972-9
Supported by: Conselho Nacional de Desenvolvimento Científico e Tecnológico 401452/2016-6
Supported by: Conselho Nacional de Desenvolvimento Científico e Tecnológico 406542/2013-9
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Abstract

Proteases play a pivotal role in many signaling pathways; inhibitors of well-established proteases have shown a substantial therapeutic success. This study aimed to examine the in vivo effects of 3 protease inhibitors isolated from Bauhinia species: i) Bauhinia mollis elastase inhibitor, which blocks human neutrophil elastase (Kiapp 2.8 nM) and cathepsin G (Kiapp 1.0 nM) activities; ii) Bauhinia mollis trypsin inhibitor, a trypsin inhibitor (Kiapp 5.0 nM); and iii) Bauhinia bauhinioides cruzipain inhibitor, which inhibits elastase (Kiapp 2.6 nM), cathepsin G (Kiapp 160.0 nM), and the cysteine proteases cathepsin L (Kiapp 0.2 nM). Bauhinia bauhinioides cruzipain inhibitor, Bauhinia mollis elastase inhibitor, and Bauhinia mollis trypsin inhibitor were isolated using acetone and ammonium sulfate fractionations, DEAE-Sephadex, trypsin-Sepharose, and Resource-Q chromatographies. Mice and rats were treated intraperitoneally with 1 dose of inhibitor; gastric mucosal lesions were induced by cold-restraint stress. Oral pretreatment of mice with Bauhinia mollis elastase inhibitor or Bauhinia mollis trypsin inhibitor (1 – 10 mg/kg) did not show anti-ulcer effect, while Bauhinia bauhinioides cruzipain inhibitor (0.1 – 1.0 mg/kg) produced a similar reduction of the index of mucosal damage at all effective doses (30 to 33% < control). In rats at doses lower than those used in mice, Bauhinia mollis elastase inhibitor and Bauhinia bauhinioides cruzipain inhibitor reduced the index of mucosal damage by 66% and 54% of controls, respectively. The results indicate a protective effect against gastric mucosal lesions associated with elastase inhibition but not inhibition of trypsin activities. Moreover, the lack of Bauhinia mollis elastase inhibitor efficacy observed in mice may possibly be related to the reported structural differences of elastase in mice and rats.

Key words

Bauhinia bauhiniodes - Bauhinia mollis - elastase - gastric mucosal lesions - Leguminosae - protease inhibitor

Supporting Information

    Isolation of Bauhinia mollis elastase inhibitor (BmEI) and Bauhinia mollis trypsin inhibitor (BmTI).

  • Supporting Information


Publication History

Received: 09 April 2020

Accepted after revision: 13 June 2020

Article published online:
14 July 2020

© 2020. Thieme. All rights reserved.

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  • References

  • 1 Pott A, Pott VJ. Plants of Pantanal – Embrapa – Centro de Pesquisa Agropecuaria do Pantanal (Corumba, MS). Brasilia: EMBRAPA-SPl; 1994
    Google Scholar
  • 2 Mello VJ, Gomes MTR, Lemos FO, Delfino JL, Andrade SP, Lopes MTP, Salas CE. The gastric ulcer protective and healing role of cysteine proteinases from Carica candamarcensis . Phytomedicine 2008; 15: 237-244
    CrossrefPubMedGoogle Scholar
  • 3 Bhalke RD, Giri MA, Pal SC. Antiulcer activity of methanol extract of leaves of Bauhinia racemosa Linn (Leguminaceae). Research J Pharm and Tech 2010; 4: 124-127
    PubMedGoogle Scholar
  • 4 Chechinel Filho V. Chemical composition and biological potential of plants from the genus Bauhinia . Phytother Res 2009; 23: 1347-1354
    CrossrefPubMedGoogle Scholar
  • 5 Oliva MLV, Silva MCC, Sallai RC, Brito MV, Sampaio MU. A novel subclassification for Kunitz proteinase inhibitors from leguminous seeds. Biochimie 2010; 92: 1667-1673
    CrossrefPubMedGoogle Scholar
  • 6 Almeida-Reis R, Theodoro-Junior OA, Oliveira BTM, Oliva LV, Toledo-Arruda AC, Bonturi CR, Brito MV, Lopes FDTQS, Prado CM, Florencio AC, Martins MA, Owen CA, Leick EA, Oliva MLV, Tibério IFLC. Plant proteinase inhibitor BbCI modulates lung inflammatory responses and mechanic and remodeling alterations induced by elastase in mice. Biomed Res Int 2017; 2017: 1-13
    CrossrefPubMedGoogle Scholar
  • 7 Rodrigues APD, Bortolozzo ASS, Arantes-Costa FM, Saraiva-Romanholo BM, de Souza FCR, Brüggemann TR, Santana FPR, de Brito MV, Bonturi CR, Nunes NNDS, Prado CM, Leick EA, Oliva MLV, Martins MA, Righetti RF, Tibério IFLC. A plant proteinase inhibitor from Enterolobium contortisiliquum attenuates airway hyperresponsiveness, inflammation and remodeling in a mouse model of asthma. Histol Histopathol 2019; 34: 537-552
    PubMedGoogle Scholar
  • 8 Oliva MLV, Sampaio MU. Bauhinia Kunitz-type proteinase inhibitors: structural characteristics and biological properties. Biol Chem 2008; 389: 1007-1013
    CrossrefPubMedGoogle Scholar
  • 9 de Oliveira C, Santana LA, Carmona AK, Cezari MH, Sampaio MU, Sampaio CAM, Oliva MLV. Structure of Cruzipain/Cruzain inhibitors isolated from Bauhinia bauhinioides seeds. Biol Chem 2001; 382: 847-852
    CrossrefPubMedGoogle Scholar
  • 10 Araújo APU, Hansen D, Vieira DF, de Oliveira C, Santana LA, Beltramini LM, Sampaio CAM, Sampaio MU, Oliva MLV. Kunitz-type Bauhinia bauhinioides inhibitors devoid of disulfide bridges: isolation of the cDNAs, heterologous expression and structural studies. Biol Chem 2005; 386: 561-568
    PubMedGoogle Scholar
  • 11 Oliveira C, Navarro-Xavier RA, Anjos-Vallota EA, Martins JO, Silveira VLF, Gonçalves LRC, Araújo MS, Motta G, Sannomiya P, Oliva MLV. Effect of plant neutrophil elastase inhibitor on leucocyte migration, adhesion and cytokine release in inflammatory conditions. Br J Pharmacol 2010; 161: 899-910
    CrossrefPubMedGoogle Scholar
  • 12 Kushimoto S, Okajima K, Okabe H, Binder BR. Role of granulocyte elastase in the formation of hemorrhagic shock-induced gastric mucosal lesions in the rat. Crit Care Med 1996; 24: 1041-1046
    CrossrefPubMedGoogle Scholar
  • 13 Liu W, Okajima K, Murakami K, Harada N, Isobe H, Irie T. Role of neutrophil elastase in stress-induced gastric mucosal injury in rats. J Lab Clin Med 1998; 132: 432-439
    CrossrefPubMedGoogle Scholar
  • 14 Murakami K, Okajima K, Uchiba M, Harada N, Liu W, Okabe H, Takatsuki K. Role of granulocyte elastase in indomethacininduced gastric mucosal lesion formation in rats. J Lab Clin Med 1997; 130: 307-313
    CrossrefPubMedGoogle Scholar
  • 15 Zimmerman BJ, Granger DN. Reperfusion-induced leukocyte infiltration: Role of elastase. Am J Physiol Circ Physiol 1990; 259: H390-394
    CrossrefPubMedGoogle Scholar
  • 16 Okajima K, Murakami K, Liu W, Uchiba M. Inhibition of neutrophil activation by ranitidine contributes to prevent stress-induced gastric mucosal injury in rats. Crit Care Med 2000; 28: 2858-2865
    CrossrefPubMedGoogle Scholar
  • 17 Harada N, Okajima K, Murakami K, Isobe H, Liu W. Gastric prostacyclin (PGI2) prevents stress-induced gastric mucosal injury in rats primarily by inhibiting leukocyte activation. Prostaglandins Other Lipid Mediat 1999; 57: 291-303
    CrossrefPubMedGoogle Scholar
  • 18 Kessenbrock K, Dau T, Jenne DE. Tailor-made inflammation: how neutrophil serine proteases modulate the inflammatory response. J Mol Med 2011; 89: 23-28
    CrossrefPubMedGoogle Scholar
  • 19 Korkmaz B, Horwitz MS, Jenne DE, Gauthier F. Neutrophil elastase, proteinase 3, and cathepsin G as therapeutic targets in human diseases. Pharmacol Rev 2010; 62: 726-759
    CrossrefPubMedGoogle Scholar
  • 20 Scott CJ, Taggart CC. Biologic protease inhibitors as novel therapeutic agents. Biochimie 2010; 92: 1681-1688
    CrossrefPubMedGoogle Scholar
  • 21 Oliva MLV, Sampaio MU. Action of plant proteinase inhibitors on enzymes of physiopathological importance. An Acad Bras Cienc 2009; 81: 615-621
    CrossrefPubMedGoogle Scholar
  • 22 Sumikawa JT, Nakahata AM, Fritz H, Mentele R, Sampaio MU, Oliva MLV. A Kunitz-type glycosylated elastase inhibitor with one disulfide bridge. Planta Med 2006; 72: 393-397
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Woloszynek JC, Hu Y, Pham CTN. Cathepsin G-regulated release of formyl peptide receptor agonists modulate neutrophil effector functions. J Biol Chem 2012; 287: 34101-34109
    CrossrefPubMedGoogle Scholar
  • 24 Bonaventura A, Liberale L, Carbone F, Vecchié A, Diaz-Cañestro C, Camici GG, Montecucco F, Dallegri F. The pathophysiological role of neutrophil extracellular traps in inflammatory diseases. Thromb Haemost 2018; 118: 6-27
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Harada N, Okajima K, Murakami K, Isobe H, Liu W. Gastric prostacyclin (PGI2) prevents stress-induced gastric mucosal injury in rats primarily by inhibiting leukocyte activation. Prostaglandins Other Lipid Mediat 1999; 57: 291-303
    CrossrefPubMedGoogle Scholar
  • 26 Caruso R, Fina D, Peluso I, Fantini MC, Tosti C, Blanco GDV, Paoluzi OA, Caprioli F, Andrei F, Stolfi C, Romano M, Ricci V, MacDonald TT, Pallone F, Monteleone G. IL-21 is highly produced in Helicobacter pylori-infected gastric mucosa and promotes gelatinases synthesis. J Immunol 2007; 178: 5957-5965
    CrossrefPubMedGoogle Scholar
  • 27 Sampieri CL, de la Peña S, Ochoa-Lara M, Zenteno-Cuevas R, León-Córdoba K. Expression of matrix metalloproteinases 2 and 9 in human gastric cancer and superficial gastritis. World J Gastroenterol 2010; 16: 1500-1505
    CrossrefPubMedGoogle Scholar
  • 28 Bulgari M, Sangiovanni E, Colombo E, Maschi O, Caruso D, Bosisio E, DellʼAgli M. Inhibition of neutrophil elastase and metalloprotease-9 of human adenocarcinoma gastric cells by chamomile (Matricaria recutita L.) infusion. Phytother Res 2012; 26: 1817-1822
    CrossrefPubMedGoogle Scholar
  • 29 Regoli D, Gobeil F. Kinins and peptide receptors. Biol Chem 2016; 397: 297-304
    CrossrefPubMedGoogle Scholar
  • 30 Harada N, Okajima K, Liu W. Rebamipide decreases the susceptibility of gastric mucosa to acid-induced injury in rats by inhibiting neutrophil activation. Dig Dis Sci 2005; 50: 56-62
    CrossrefPubMedGoogle Scholar
  • 31 Puente XS. A Genomic Analysis of rat proteases and protease inhibitors. Genome Res 2004; 14: 609-622
    CrossrefPubMedGoogle Scholar
  • 32 Heinz A, Jung MC, Jahreis G, Rusciani A, Duca L, Debelle L, Weiss AS, Neubert RHH, Schmelzer CEH. The action of neutrophil serine proteases on elastin and its precursor. Biochimie 2012; 94: 192-202
    CrossrefPubMedGoogle Scholar
  • 33 Johnson D, Travis J. The oxidative inactivation of human alpha-1-proteinase inhibitor. Further evidence for methionine at the reactive center. J Biol Chem 1979; 254: 4022-4026
    CrossrefPubMedGoogle Scholar
  • 34 Beatty K, Bieth J, Travis J. Kinetics of association of serine proteinases with native and oxidized alpha-1-proteinase inhibitor and alpha-1-antichymotrypsin. J Biol Chem 1980; 255: 3931-3934
    CrossrefPubMedGoogle Scholar
  • 35 Taggart C, Cervantes-Laurean D, Kim G, McElvaney NG, Wehr N, Moss J, Levine RL. Oxidation of either methionine 351 or methionine 358 in alpha 1-antitrypsin causes loss of anti-neutrophil elastase activity. J Biol Chem 2000; 275: 27258-27265
    CrossrefPubMedGoogle Scholar
  • 36 Edman P, Högfeldt E, Sillén LG, Kinell PO. Method for determination of the amino acid sequence in peptides. Acta Chem Scand 1950; 4: 283-293
    CrossrefPubMedGoogle Scholar
  • 37 Oliva MLV, Mendes CR, Juliano MA, Chagas JR, Rosa JC, Greene LJ, Sampaio MU, Sampaio CA. Characterization of a tissue kallikrein inhibitor isolated from Bauhinia bauhinioides seeds: inhibition of the hydrolysis of kininogen related substrates. Immunopharmacology 1999; 45: 163-169
    CrossrefPubMedGoogle Scholar
  • 38 Morrison AR, Winokur TS, Brown WA. Inhibition of soybean lipoxygenase by mannitol. Biochem Biophys Res Commun 1982; 108: 1757-1762
    CrossrefPubMedGoogle Scholar
  • 39 Knight CG. The Characterization of Enzymes Inhibition. In: Barrett AJ, Salvesen G. eds. Proteinase Inhibitors. Amsterdam: Elsevier; 1986: 23-51
    Google Scholar
  • 40 Biondo TMA, Tanae MM, Coletta ED, Lima-Landman MTR, Lapa AJ, Souccar C. Antisecretory actions of Baccharis trimera (Less.) DC aqueous extract and isolated compounds: Analysis of underlying mechanisms. J Ethnopharmacol 2011; 136: 368-373
    CrossrefPubMedGoogle Scholar
  • 41 Nakahata AM, Bueno NR, Rocha HAO, Franco CRC, Chammas R, Nakaie CR, Jasiulionis MG, Nader HB, Santana LA, Sampaio MU, Oliva MLV. Structural and inhibitory properties of a plant proteinase inhibitor containing the RGD motif. Int J Biol Macromol 2006; 40: 22-29
    CrossrefPubMedGoogle Scholar