Effects of quercetin on hyper-proliferation of gastric mucosal cells in rats treated with chronic oral ethanol through the reactive oxygen species-nitric oxide pathway.
OBJECTIVE
To investigate the effect of quercetin (3,3',4',5,7-pentahydroxy flavone), a major flavonoid in human diet, on hyper-proliferation of gastric mucosal cells in rats treated with chronic oral ethanol.


METHODS
Forty male Sprague-Dawley rats, weighing 200-250 g, were randomly divided into control group (tap water ad libitum), ethanol treatment group (6 mL/L ethanol), quercetin treatment group (intragastric gavage with 100 mg/kg of quercetin per day), and ethanol plus quercetin treatment group (quercetin and 6 mL/L ethanol). Expression levels of proliferating cell nuclear antigen (PCNA) and Cyclin D1 were detected by Western blot to assay gastric mucosal cell proliferation in rats. To demonstrate the influence of quercetin on the production of extra-cellular reactive oxygen species/nitrogen species (ROS/RNS) in rats, changes in levels of thiobarbituric acid reactive substance (TBARS), protein carbonyl, nitrite and nitrate (NOx) and nitrotyrosine (NT) were determined. The activity of inducible nitric oxide synthase (NOS) including iNOS and nNOS was also detected by Western blot.


RESULTS
Compared to control animals, cell proliferation in the gastric mucosa of animals subjected to ethanol treatment for 7 days was significant increased (increased to 290% for PCNA density P < 0.05, increased to 150 for Cyclin D1 density P < 0.05 and 21.6 +/- 0.8 vs 42.3 +/- 0.7 for PCNA positive cells per view field), accompanied by an increase in ROS generation (1.298 +/- 0.135 micromol vs 1.772 +/- 0.078 micromol for TBARS P < 0.05; 4.36 +/- 0.39 mmol vs 7.48 +/- 0.40 mmol for carbonyl contents P < 0.05) and decrease in NO generation (11.334 +/- 0.467 micromol vs 7.978 +/- 0.334 micromol P < 0.01 for NOx; 8.986 +/- 1.351 micromol vs 6.854 +/- 0.460 micromol for nitrotyrosine P < 0.01) and nNOS activity (decreased to 43% P<0.05). This function was abolished by the co-administration of quercetin.


CONCLUSIONS
The antioxidant action of quercetin relies, in part, on its ability to stimulate nNOS and enhance production of NO that would interact with endogenously produced reactive oxygen to inhibit hyper-proliferation of gastric mucosal cells in rats treated with chronic oral ethanol.
Liu
Jing-Li
JL
Department of Physiology, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China.
Du
Jun
J
Fan
Ling-Ling
LL
Liu
Xiao-Yan
XY
Gu
Luo
L
Ge
Ying-Bin
YB
D000284:Administration, Oral; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects*; D056741:Cyclin D; D016213:Cyclins / Q000378:metabolism; D000431:Ethanol / Q000008:administration & dosage / Q000633:toxicity*; D005419:Flavonoids / Q000494:pharmacology*; D005753:Gastric Mucosa / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D050050:Protein Carbonylation / Q000187:drug effects; D011794:Quercetin; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D000975:Antioxidants; D056741:Cyclin D; D016213:Cyclins; D005419:Flavonoids; D018809:Proliferating Cell Nuclear Antigen; D017382:Reactive Oxygen Species; D017392:Thiobarbituric Acid Reactive Substances; D009569:Nitric Oxide; C002744:3-nitrotyrosine; D000431:Ethanol; D014443:Tyrosine; D011794:Quercetin; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; C496322:Nos1 protein, rat