title: Evidence of gastric ulcer healing activity of Maytenus robusta Reissek: In vitro and in vivo studies.
abstract: BACKGROUND
  Maytenus robusta Reissek (Celastraceae) is traditionally used in Brazilian folk medicine to treat gastric ulcer, as a substitute for M. ilicifolia, which is almost extinct. The gastroprotective properties of M. robusta were demonstrated previously using only preventive approaches, such as acute gastric ulcer models. However, the healing effect of M. robusta in gastric ulcers remains unclear.
  
  
  OBJECTIVE
  The current study was carried out to investigate the healing effectiveness of M. robusta hydroalcoholic extract (HEMR) from aerial parts in the acetic acid-induced chronic ulcer model and to determine its effect on cell proliferation, scavenging free radicals, and inflammatory and oxidative damage.
  
  
  METHODS
  To evaluate the healing properties of HEMR in vivo, chronic gastric ulcer was induced in rats by 80% acid acetic. Next, different groups of animals (n=6) were treated orally with vehicle (water plus 1% tween, 1 ml/kg), omeprazole (20mg/kg), or HEMR (1-10mg/kg), twice daily for 7 days. At the end of the treatment, the total ulcer area (mm(2)) was measured and a sample of gastric tissue was taken for histological and histochemical analysis. Evaluation of GSH and LOOH levels, GST, SOD, CAT and MPO activity was also performed at the site of the lesion. In parallel, radical scavenging activity, cytoprotective effect, and cell proliferation activity in fibroblasts (L929 cells) were determined by in vitro trials. The antisecretory properties were evaluated using the pylorus ligature model in rats, and the anti-Helicobacter pylori activity was determined in vitro. Acute toxicity was evaluated by relative organ weight and biochemical parameters in serum. The prokinetic properties were also evaluated in mice.
  
  
  RESULTS
  Oral administration of HEMR (10mg/kg) reduced the gastric ulcer area by 53%, compared to the vehicle group (120.0 ± 8.3mm(2)), the regeneration of gastric mucosa was evidenced in histological analysis. Moreover, HEMR treatment increased gastric mucin content and reduced oxidative stress and inflammatory parameters at the site of the ulcer. In vitro, HEMR (1-1000 µg/ml) was able to scavenge free radical DPPH and promote cytoprotection against H2O2 in fibroblasts at 0.1-100 µg/ml. Moreover, HEMR healing properties also were confirmed by enhancement of proliferation and coverage of scratched wounds in fibroblast monolayer. However, HEMR (10mg/kg) by the intraduodenal route did not promote changes in volume, pH, total acidity or pepsin activity in the pylorus ligature model, and HEMR up to 2000 µg/ml also did not present considerable activity against H. pylori. In relation to gastrointestinal motility, HEMR (10mg/kg, p.o) did not provoke alterations. It is also important to mention that oral administration of HEMR did not produce any sign of acute toxicity in animals.
  
  
  CONCLUSIONS
  The data here obtained show that M. robusta has evident ulcer healing potential, mainly through the strengthening of protective factors of gastric mucosa, such as mucus layer, antioxidant defenses and cell proliferation. Taking into account the advantages of cultivation and harvesting of M. robusta compared to M. ilicifolia, and the evidence presented here, it is plausible to conclude that hydroalcoholic extract obtained from aerial parts of M. robusta is an interesting source for the development of a phytotherapeutic formulation to treat gastric ulcer.
authors.affiliation: Programa de Pós-Graduação em Ciência s Farmacêuticas (PPGCF), Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil. Electronic address: lu.isamota@hotmail.com.
authors.forename: Luisa Mota
authors.initials: LM
authors.lastname: da Silva
authors.affiliation: Programa de Pós-Graduação em Ciência s Farmacêuticas (PPGCF), Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil.
authors.forename: Thaise
authors.initials: T
authors.lastname: Boeing
authors.affiliation: Programa de Pós-Graduação em Ciência s Farmacêuticas (PPGCF), Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil.
authors.forename: Lincon Bordignon
authors.initials: LB
authors.lastname: Somensi
authors.affiliation: Programa de Pós-Graduação em Ciência s Farmacêuticas (PPGCF), Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil.
authors.forename: Benhur Judah
authors.initials: BJ
authors.lastname: Cury
authors.affiliation: Programa de Pós-Graduação em Ciência s Farmacêuticas (PPGCF), Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil.
authors.forename: Viviane Miranda Bispo
authors.initials: VM
authors.lastname: Steimbach
authors.affiliation: Departamento de Ciências Farmacêuticas, Universidade Regional de Blumenau (FURB), Blumenau, SC, Brazil.
authors.forename: Alessandro Conrado de Oliveira
authors.initials: AC
authors.lastname: Silveria
authors.affiliation: Programa de Pós-Graduação em Ciência s Farmacêuticas (PPGCF), Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil.
authors.forename: Rivaldo
authors.initials: R
authors.lastname: Niero
authors.affiliation: Programa de Pós-Graduação em Ciência s Farmacêuticas (PPGCF), Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil.
authors.forename: Valdir
authors.initials: V
authors.lastname: Cechinel Filho
authors.affiliation: Programa de Pós-Graduação em Ciência s Farmacêuticas (PPGCF), Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil.
authors.forename: José Roberto
authors.initials: JR
authors.lastname: Santin
authors.affiliation: Programa de Pós-Graduação em Ciência s Farmacêuticas (PPGCF), Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil.
authors.forename: Sérgio Faloni
authors.initials: SF
authors.lastname: de Andrade
mesh_terms: D019342:Acetic Acid; D000818:Animals; D000897:Anti-Ulcer Agents / Q000494:pharmacology / Q000627:therapeutic use*; D001938:Brazil; D002374:Catalase / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D005347:Fibroblasts / Q000187:drug effects; D005753:Gastric Mucosa / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D016480:Helicobacter pylori / Q000187:drug effects; D029755:Maytenus; D008519:Medicine, Traditional; D051379:Mice; D009195:Peroxidase / Q000378:metabolism; D008517:Phytotherapy; D035261:Plant Components, Aerial; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use*; D017208:Rats, Wistar; D013276:Stomach Ulcer / Q000139:chemically induced / Q000188:drug therapy* / Q000473:pathology; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D000897:Anti-Ulcer Agents; D010936:Plant Extracts; D002374:Catalase; D009195:Peroxidase; D013482:Superoxide Dismutase; D005982:Glutathione Transferase; D005978:Glutathione; D019342:Acetic Acid
keywords: Antioxidant; Antiulcer; Fibroblast; Gastroprotection; Healing ulcer; M. robusta