title: Gastroprotective activity of ethyl-4-[(3,5-di-tert-butyl-2-hydroxybenzylidene) amino]benzoate against ethanol-induced gastric mucosal ulcer in rats.
abstract: BACKGROUND
  The study was carried out to determine the cytotoxic, antioxidant and gastro-protective effect of ethyl-4-[(3,5-di-tert-butyl-2-hydroxybenzylid ene)amino] benzoate (ETHAB) in rats.
  
  
  RESULTS
  The cytotoxic effect of ETHAB was assessed using a MTT cleavage assay on a WRL68 cell line, while its antioxidant activity was evaluated in vitro. In the anti-ulcer study, rats were divided into six groups. Group 1 and group 2 received 10% Tween 20 (vehicle). Group 3 received 20 mg/kg Omeprazole. Groups 4, 5 and 6 received ETHAB at doses of 5, 10, and 20 mg/kg, respectively. After an hour, group 1 received the vehicle. Groups 2-6 received absolute ethanol to induce gastric mucosal lesions. In the WRL68 cell line, an IC50 of more than 100 µg/mL was observed. ETHAB results showed antioxidant activity in the DPPH, FRAP, nitric oxide and metal chelating assays. There was no acute toxicity even at the highest dosage (1000 mg/kg). Microscopy showed that rats pretreated with ETHAB revealed protection of gastric mucosa as ascertained by significant increases in superoxide dismutase (SOD), pH level, mucus secretion, reduced gastric lesions, malondialdehyde (MDA) level and remarkable flattened gastric mucosa. Histologically, pretreatment with ETHAB resulted in comparatively better gastric protection, due to reduction of submucosal edema with leucocyte infiltration. PAS staining showed increased intensity in uptake of Alcian blue. In terms of immunohistochemistry, ETHAB showed down-expression of Bax proteins and over-expression of Hsp70 proteins.
  
  
  CONCLUSIONS
  The gastroprotective effect of ETHAB may be attributed to antioxidant activity, increased gastric wall mucus, pH level of gastric contents, SOD activity, decrease in MDA level, ulcer area, flattening of gastric mucosa, reduction of edema and leucocyte infiltration of the submucosal layer, increased PAS staining, up-regulation of Hsp70 protein and suppressed expression of Bax.
authors.affiliation: Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia; Al-Moalim Mohamed Awad Center for Scientific Miracles of Prophetic Medicine, College of Medicine, Taibah University, Al-Madinah Al-Munawarah, Saudi Arabia.
authors.forename: Mohammed Farouq
authors.initials: MF
authors.lastname: Halabi
authors.affiliation: Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia; Department. of Chemistry, Ibn Al-Haitham, University of Baghdad. Baghdad, Iraq.
authors.forename: Raied Mustafa
authors.initials: RM
authors.lastname: Shakir
authors.affiliation: Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Daleya Abdulaziz
authors.initials: DA
authors.lastname: Bardi
authors.affiliation: Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Nahla Saeed
authors.initials: NS
authors.lastname: Al-Wajeeh
authors.affiliation: Institute of Biological Science, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Abdulwali
authors.initials: A
authors.lastname: Ablat
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Pouya
authors.initials: P
authors.lastname: Hassandarvish
authors.affiliation: Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Maryam
authors.initials: M
authors.lastname: Hajrezaie
authors.affiliation: Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Anwar
authors.initials: A
authors.lastname: Norazit
authors.affiliation: Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Mahmood Ameen
authors.initials: MA
authors.lastname: Abdulla
mesh_terms: D000818:Animals; D000897:Anti-Ulcer Agents / Q000138:chemical synthesis / Q000494:pharmacology / Q000627:therapeutic use*; D000975:Antioxidants / Q000138:chemical synthesis / Q000494:pharmacology / Q000627:therapeutic use*; D000431:Ethanol / Q000633:toxicity; D005260:Female; D005753:Gastric Mucosa / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D008297:Male; D010636:Phenols / Q000494:pharmacology / Q000627:therapeutic use*; D051381:Rats; D013276:Stomach Ulcer / Q000188:drug therapy*; D062366:para-Aminobenzoates / Q000494:pharmacology / Q000627:therapeutic use*
chemical_list: D000897:Anti-Ulcer Agents; D000975:Antioxidants; D010636:Phenols; C000601991:ethyl 4-((3,5-di-tert-butyl-2-hydroxybenzylidene)amino)benzoate; D062366:para-Aminobenzoates; D000431:Ethanol