Pioglitazone, a specific ligand of the peroxisome proliferator-activated receptor gamma reduces gastric mucosal injury induced by ischaemia/ reperfusion in rat.
BACKGROUND
The peroxisome proliferator-activated receptor gamma (PPARgamma) is a ligand-dependent nuclear receptor that has been implicated in the control of metabolism and numerous cellular processes, including cell cycle control, carcinogenesis, and inflammation. The present study was designed to investigate the effect of the specific PPARgamma ligand, pioglitazone, on the mucosal lesions induced by ischaemia and reperfusion (I/R) in rats.


METHODS
I/R lesions were induced in Wistar rats by applying a small clamp to the coeliac artery for 30 min (ischaemic phase), followed by the removal of the clamp for 3 h (reperfusion phase). Vehicle (saline) or increasing doses of pioglitazone (2.5, 10, and 30 mg/kg i.g.) were given 30 min before exposure to I/R. The animals were killed immediately after the end of the reperfusion phase (time 0) and at 12 and 24h after I/R. The area of gastric lesions was measured by planimetry, and the gastric blood flow was determined by the H2 gas clearance method. The gastric mucosal gene expressions of PPARgamma, interleukin-1beta (IL-1beta), tumour necrosis factor alpha (TNF-alpha), leptin, cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) were examined by RT-PCR. In addition, protein expression of COX-2 and leptin was assessed by Western blot.


RESULTS
The pretreatment with pioglitazone reduced in a dose-dependent manner the mean lesion area induced by I/R, and this effect was accompanied by a significant increase in the gastric blood flow. The decrease in gastric ulcerations by pioglitazone was also observed 12 and 24 h after the I/R. The PPARgamma mRNA was weakly expressed in the intact gastric mucosa but significantly up-regulated after exposure to I/R at each time interval studied. The expression of IL-1beta was not changed significantly after pioglitazone applied i.g. at doses 2.5 and 10 mg/kg, but it was down-regulated at the dose 30mg/kg. TNFalpha mRNA was strongly increased after the exposure to I/R, but it was down-regulated after pioglitazone pretreatment. In contrast, both leptin and COX-2 mRNA and protein expression were increased in the gastric mucosa after exposure to I/R. The pretreatment with pioglitazone caused a significant up-regulation of mRNA and protein expression of leptin, reaching its peak at the dose 30 mg/kg i.g. In contrast, COX-2 expression did not change significantly after the 2.5 and 10 mg/kg of pioglitazone, but it significantly decreased after pioglitazone at dose 30 mg/kg given to rats before exposure to I/R.


CONCLUSIONS
Pioglitazone reduces the acute erosions and deeper gastric lesions induced by I/R. The beneficial effect of this PPARgamma ligand on I/R-induced gastric damage may be due to its anti-inflammatory properties, especially to the reduction in TNF-alpha expression and to up-regulation of leptin mRNA in the gastric mucosa. The inhibition of COX-2 expression by pioglitazone may reflect the anti-inflammatory properties of this compound.
Konturek
P C
PC
Dept. of Medicine I, University Erlangen-Nuremberg, Erlangen, Germany. pkonturek@aol.com
Brzozowski
T
T
Kania
J
J
Kukharsky
V
V
Bazela
K
K
Kwiecien
S
S
Harsch
I
I
Konturek
S J
SJ
Hahn
E G
EG
D000595:Amino Acid Sequence; D000818:Animals; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D005753:Gastric Mucosa / Q000098:blood supply / Q000187:drug effects* / Q000378:metabolism*; D007004:Hypoglycemic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D007375:Interleukin-1 / Q000096:biosynthesis; D007527:Isoenzymes / Q000096:biosynthesis; D020738:Leptin / Q000096:biosynthesis; D008297:Male; D008565:Membrane Proteins; D023421:Models, Animal; D000077205:Pioglitazone; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D015427:Reperfusion Injury / Q000150:complications / Q000517:prevention & control*; D013152:Splanchnic Circulation / Q000187:drug effects*; D013276:Stomach Ulcer / Q000209:etiology / Q000517:prevention & control*; D013844:Thiazoles / Q000494:pharmacology* / Q000627:therapeutic use; D045162:Thiazolidinediones; D014157:Transcription Factors / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D007004:Hypoglycemic Agents; D007375:Interleukin-1; D007527:Isoenzymes; D020738:Leptin; D008565:Membrane Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D013844:Thiazoles; D045162:Thiazolidinediones; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; C496513:Ptgs1 protein, rat; D000077205:Pioglitazone