Korean J Physiol Pharmacol.  2013 Feb;17(1):81-87. 10.4196/kjpp.2013.17.1.81.

Anti-Oxidative and Anti-Inflammatory Effects of QGC in Cultured Feline Esophageal Epithelial Cells

• Lee MJ
• Song HJ
• Jeong JY
• Park SY
• Sohn UD

• KMID: 1432767
• DOI: http://doi.org/10.4196/kjpp.2013.17.1.81

Abstract

Quercetin-3-O-beta-D-glucuronopyranoside (QGC) is a flavonoid glucoside extracted from Rumex Aquaticus Herba. In the present study, anti-oxidative and anti-inflammatory effects of QGC were tested in vitro. Epithelial cells obtained from cat esophagus were cultured. When the cells were exposed to acid for 2 h, cell viability was decreased to 36%. Pretreatment with 50 microM QGC for 2 h prevented the reduction in cell viability. QGC also inhibited the productions of intracellular ROS by inflammatory inducers such as acid, lipopolysaccharide, indomethacin and ethanol. QGC significantly increased the activities of superoxide dismutase (SOD) and catalase, and also induced the expression of SOD2, while it restored the decrease of catalase expression in cells exposed to acid. QGC inhibited NF-kappaB translocation, cyclooxygenase-2 expression and PGE2 secretion in cells exposed to acid, which plays an important role in the pathogenesis of esophagitis. The data suggest that QGC may well be one of the promising substances to attenuate oxidative epithelial cell injury and inflammatory signaling in esophagus inflammation.

Keyword

NF-kappaB; PGE2; QGC; ROS

MeSH Terms

Animals
Catalase
Cats
Cell Survival
Cyclooxygenase 2
Dinoprostone
Epithelial Cells
Esophagitis
Esophagus
Ethanol
Indomethacin
Inflammation
NF-kappa B
Quercetin
Rumex
Superoxide Dismutase
Catalase
Cyclooxygenase 2
Dinoprostone
Ethanol
Indomethacin
NF-kappa B
Quercetin
Superoxide Dismutase

Figure

• 

  Fig. 1 Cell viability evaluated by MTT assay. (A) Effect of QGC on cell viability of esophageal epithelial cells. (B) Time course of acid (pH medium)-induced cell injury. (C) Time course analyses for effect of QGC on acid-induced cell damage. (D) Dose-response analyses for effect of QGC on acid-induced cell damage. Serum-starved cells were pre-incubated in the presence or absence of QGC for the indicated time, and then stimulated with or without acid. Data are expressed as means±SEM of three independent experiments (Student's t-test: *p<0.05, ***p<0.001 vs. control; ###p<0.001 vs. cells in pH 4 medium alone).

• 

  Fig. 2 Effect of QGC on intracellular H2O2 level in cells exposed to acid. Serum-starved cells were pre-incubated in the presence of 50 µM QGC for 2 h. Cells were then stimulated with the indicated inflammatory inducers: (A) pH 4 medium for 10 min (B) 1 µM/ml LPS 15 h (C) 2 mM indomethacin for 2 h and (D) 10% ethanol for 1 h. ROS production was estimated using DCF-DA assay. Data are expressed as means±SEM of three experiments (Student's t-test: *p<0.05 vs. control; #p<0.05, ##p<0.01, ###p<0.001 vs. cells in inflammatory inducer alone).

• 

  Fig. 3 Effect of QGC on activities and expressions of SOD and catalase in cells exposed to acid. (A) Increase of SOD activity induced by QGC. (B) Induction of SOD2 caused by QGC. (C) Increase of catalase activity induced by QGC. (D) Restoration of catalase expression induced by QGC. Data are expressed as means±SEM of three experiments (Student's t-test: *p<0.05 vs. control; #p<0.05, ##p<0.01, ###p<0.001 vs. cells in pH 4 medium alone).

• 

  Fig. 4 Inhibitory effects of QGC on acid-induced NF-κB translocation in cells. Serum starved cells were pre-incubated in the presence of 50 µM QGC for the indicated time. Cells were then stimulated with acidic medium for 10 min. NF-κB expression was estimated by western blot analysis. Densitometric assessments demonstrated a time-dependent inhibitory effect of QGC on acid-induced NF-κB translocation. Data are expressed as means±SEM of three experiments (Student's t-test; **p<0.01 vs. control, ##p<0.01 vs. cells in acidic pH 4 medium alone).

• 

  Fig. 5 Inhibitory effects of QGC on COX2 expression and PGE2 secretion in cells exposed to acid. Serum-starved cells were pre- incubated in the presence of 50 µM QGC for the indicated time. Cells were then stimulated with acidic medium for 10 min. (A) COX2 expression was estimated by western blotting. (B) PGE2 secretion was estimated by EIA kit. Data are expressed as means±SEM of three experiments (Student's t-test: *p<0.05, **p<0.01 vs. control; #p<0.05, ##p<0.01vs. cells in acidic pH 4 medium alone).

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