Tryptophan free diet delays healing of chronic gastric ulcers in rat.
Melatonin (MT) is an ubiquitous molecule, representing one of the phylogenetically oldest signaling mechanisms. Our previous studies demonstrated that MT and its precursor L-tryptophan (L-Trp) show strong protective effect on gastric mucosa. The aim of the present study was: 1) to assess the effect of MT and L-Trp on healing of chronic gastric ulcer and accompanying changes in gastric mucosal blood flow (GBF); 2) to study the effect of MT and L-Trp on expression of iNOS. cNOS and HSP70 in ulcerated mucosa; 3) to compare the effect of L-Trp free and L-Trp rich diet on ulcer healing and gene expression of proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha), superoxide dismutase (SOD), cyclooxygenase-2 (COX-2) and NFkappaB-p65 protein expression in ulcer area and intact non-ulcerated. Chronic ulcers were induced in Wistar rats by Okabe's modification of acetic acid method. Rats with chronic gastric ulcers were divided in following treatment groups: 1) vehicle (saline); 2) MT (20mg/kg-d i.p.) and 3) L-Trp (100 mg/kg i.p.). The expression of iNOS, cNOS and HSP70 protein was measured by Western blot. In separate experiments, the influence of commercially available (Bio-Serv, USA) L-Trp free diet (TFD) was compared to the L-Trp rich diet (TRD) on the course of ulcer healing was assessed. The ulcer area was measured by planimetry. The expression of TNFalpha, COX-2 and SOD mRNA in ulcerated mucosa was analyzed by RT-PCR method. MT and its precursor L-Trp significantly accelerated ulcer healing. Healing ulcerated mucosa showed increased protein expression of iNOS and HSP70 as compared to intact gastric mucosa. TFD in contrast to normal diet significantly attenuated the ulcer healing, whereas the TRD exerted opposite effects and significantly accelerated ulcer healing. This last effect was accompanied by significant decrease of TNF-alpha mRNA expression and expression of NFkB-p65 in gastric mucosa. We conclude that: 1) MT and its precursor L-Trp significantly accelerate healing of gastric ulcer; 2) L-Trp free diet significantly attenuates experimental ulcer healing and this is due to decreased synthesis of MT from L-Trp by EE cells in gastric mucosa and 3) MT shows strong anti-inflammatory effects due to inhibition of NFkappaB and TNF-alpha expression.
Konturek
P C
PC
First Department of Medicine, University Erlangen-Nuremberg, Erlangen, Germany. Peter.Konturek@uk-erlangen.de
Burnat
G
G
Brzozowski
T
T
Zopf
Y
Y
Konturek
S J
SJ
D000818:Animals; D000897:Anti-Ulcer Agents / Q000494:pharmacology*; D015153:Blotting, Western; D002908:Chronic Disease; D051546:Cyclooxygenase 2 / Q000378:metabolism; D004032:Diet; D005753:Gastric Mucosa / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000096:biosynthesis; D008297:Male; D008550:Melatonin / Q000378:metabolism / Q000494:pharmacology; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013276:Stomach Ulcer / Q000188:drug therapy* / Q000378:metabolism; D014364:Tryptophan / Q000172:deficiency* / Q000378:metabolism / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D000897:Anti-Ulcer Agents; D006360:Heat-Shock Proteins; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D014364:Tryptophan; D019001:Nitric Oxide Synthase; D051546:Cyclooxygenase 2; D008550:Melatonin