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Role of the antioxidant properties in the gastroprotective and gastric healing activity promoted by Brazilian green propolis and the healing efficacy of Artepillin C

  • Philipe Costa
  • Lincon Bordignon Somensi
  • Rita de Cássia Melo Vilhena de Andrade Fonseca da Silva
  • Luísa Nathalia Bolda Mariano
  • Thaise Boeing
  • Bruna Longo
  • Ellen Perfoll
  • Priscila de Souza
  • Lucas Fernando Sérgio Gushiken
  • Cláudia Helena Pellizzon
  • Débora Munhoz Rodrigues
  • Jairo Kenupp Bastos
  • Sérgio Faloni de Andrade
  • Luísa Mota da SilvaEmail author
Original Article
  • 40 Downloads

Abstract

Green propolis is a resinous substance used in folk medicine given its anti-inflammatory, antibacterial, and anti-ulcer effects. Our research group has already confirmed the gastroprotective activity of hydroalcoholic extract from green propolis (HEGP), as well as of its main isolated compounds. In continuity, this study evaluated the antioxidant mode of action involved in the preventive effect induced by HEGP, and its therapeutic gastric healing potential on installed ulcers. In addition, the healing effect of its main compound Artepillin C was also investigated. Acute and chronic ulcers were induced in rats by given ethanol or acetic acid, respectively. In acute model, the rats were orally pre-treated with vehicle (water plus 1% Tween, 1 mL/kg), HEGP (30–300 mg/kg), or carbenoxolone (200 mg/kg) 1 h prior the ulcer induction. In the chronic ulcer protocol, the rats received vehicle (water plus 1% Tween, 1 mL/kg), HEGP (300 mg/kg), or omeprazole (20 mg/kg) twice a day by 7 days, whereas groups of mice received vehicle (water plus 1% Tween, 1 mL/kg), Artepillin C (18 mg/kg), or ranitidine (20 mg/kg) twice a day by 4 days. Ulcerated tissue was collected for histological, histochemical, immunostaining, oxidative, and inflammatory analyses. The in vitro scavenger activity of HEGP was also verified using the DPPH assay. The oral pre-treatment with HEGP (100 and 300 mg/kg) prevented the gastric epithelial damage promoted by ethanol. Besides, HEGP (100 and 300 mg/kg) reduced SOD activity about 11% and 26%, respectively, and increased the activity of GST around 20% and CAT in 80%. HEGP (300 mg/kg) also reduced the production of reactive oxygen species, as well as lipoperoxidation levels in the ethanol-ulcerated tissue. In the acetic acid-induced chronic ulcer, the daily treatment with HEGP (300 mg/kg) accelerates the healing process by 71%. In this model, HEGP normalized SOD and CAT activity and increased GST activity by 109% when compared to non-ulcerated rats. In both models, the extract administration increased the mucin PAS staining and reduced the myeloperoxidase activity at the ulcer site. Moreover, the treatment with HEGP enhanced the PCNA immunostaining, but did not alter the concentration of collagen in the acetic acid-ulcerated tissue. The extract had a direct DPPH radical-scavenging ability (LogIC50: 0.56). Besides, as expected, HPLC analysis showed Artepillin C as a major compound and its administration at 18 mg/kg also accelerated the gastric healing ulcer process in mice. Our findings confirm that HEGP displays both gastroprotective and gastric healing properties, contributing to the validation of its popular use as preventive and therapeutic approaches. These actions occur through the increase in mucin production and the reestablishment of the oxidative balance due to a reduction in gastric inflammation.

Keywords

Propolis Mucin Oxidative stress Gastric inflammation 

Abbreviations

ALFAC

85% Alcohol, 10% formaldehyde and 5% acetic acid

ART C

Artepillin C

BHT

Butylated hydroxytoluene

CAT

Catalase

CBX

Carbenoxolone

CDNB

1-Chloro-2,4-dinitrobenzene

DCFH-DA, 2′,7′

Dichlorofluorescin diacetate

DPPH

2,2-diphenyl-1-picrylhydrazyl

DMSO

N,N-dimetyl sulfoxide

DTNB

5,5-dithio-bis-(2-nitrobenzoic acid)

EDTA

Ethylenediaminetetraacetic

FOX2

Ferrous oxidation-xylenol orange

GSH

Reduced glutathione

GST

Glutathione-S-transferase

HEGP

Hydroalcoholic extract from green propolis

HTAB

Hexadecyltrimethylammonium bromide

IBPs

Proton pump inhibitors

LOOH

Lipoperoxides

MPO

Myeloperoxidase

OME

Omeprazole

PAS

Periodic acid of Schiff

PBS

Phosphate-buffered saline

PCNA

Proliferating cell nuclear antigen

RAN

Ranitidine

ROS

Reactive oxygen species

SOD

Superoxide dismutase

TMB

3,3′,5,5′-Tetramethylbenzidine

VEH

Vehicle

Notes

Acknowledgements

This study was financed in part by the Coordination of Improvement of Higher Education Personnel (CAPES, Portuguese: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil; CAPES)—Finance Code 001 and São Paulo Research Foundation (FAPESP, Portuguese: Fundação de Amparo à Pesquisa do Estado de São Paulo)—Finance Code 2017/04138. For the accomplishment of this study, we also counted on the assistance of Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Universidade do Vale do Itajaí (UNIVALI). Dr. Rita de Cássia Melo Vilhena de Andrade Fonseca da Silva received a Postdoctoral scholarship from PNPD/CAPES.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Philipe Costa
    • 1
    • 2
    • 3
  • Lincon Bordignon Somensi
    • 1
  • Rita de Cássia Melo Vilhena de Andrade Fonseca da Silva
    • 1
  • Luísa Nathalia Bolda Mariano
    • 1
  • Thaise Boeing
    • 1
  • Bruna Longo
    • 1
    • 2
    • 3
  • Ellen Perfoll
    • 1
  • Priscila de Souza
    • 1
  • Lucas Fernando Sérgio Gushiken
    • 2
  • Cláudia Helena Pellizzon
    • 2
  • Débora Munhoz Rodrigues
    • 3
  • Jairo Kenupp Bastos
    • 3
  • Sérgio Faloni de Andrade
    • 1
  • Luísa Mota da Silva
    • 1
    Email author
  1. 1.Programa de Pós-Graduação em Ciências Farmacêuticas, Núcleo de Investigações Químico-Farmacêuticas (NIQFAR)Universidade do Vale do Itajaí (UNIVALI)Santa CatarinaBrazil
  2. 2.Departamento de MorfologiaUniversidade do Estado de São Paulo (Unesp), Instituto de Biociências, BotucatuSão PauloBrazil
  3. 3.Faculdade de Ciências Farmacêuticas de Ribeirão PretoUniversidade de São Paulo (USP)Ribeirão PretoBrazil

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