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, 2016, 2052180

Antioxidant and Anti-Inflammatory Effects of Rhei Rhizoma and Coptidis Rhizoma Mixture on Reflux Esophagitis in Rats

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Antioxidant and Anti-Inflammatory Effects of Rhei Rhizoma and Coptidis Rhizoma Mixture on Reflux Esophagitis in Rats

O Jun Kwon et al. Evid Based Complement Alternat Med.

Abstract

The purpose of this study was to investigate the antioxidant and anti-inflammatory effects of the combined extract of Rhei rhizoma and Coptidis rhizoma (RC-mix) in experimental model of acute reflux esophagitis. The antioxidant activity was assessed by in vitro 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. RC-mix was given at 100, 200, and 400 mg/kg body weight 2 h prior to induction of reflux esophagitis (RE). After 5 h, the effects of RC-mix treated rats were compared with those of normal and control rats. The representative flavonoid contents of RC-mix, such as sennoside A, epiberberine, coptisine, palmatine, and berberine, were detected using HPLC. The elevated esophageal mucosa damage was markedly ameliorated by RC-mix treatment in a dose-dependent manner. Furthermore, the administration of RC-mix reduced the increase of serum reactive oxygen species (ROS) and peroxynitrite (ONOO(-)). The improvement of superoxide dismutase (SOD) and heme oxygenase-1 (HO-1) levels were marked in the group given RC-mix. Moreover, the elevation of inflammatory mediators and cytokines by nuclear factor-kappa B (NF-κB) activation in control rats decreased by RC-mix pretreatment. These results indicate that RC-mix treatment reduces the pathological states of esophagitis via regulating NF-κB mediated inflammation related to oxidative stress.

Figures

Figure 1
Figure 1
HPLC chromatogram of RC-mix extract (1 mg/mL) detected 254 nm. Signals 1–5 identified to be sennoside A, epiberberine, coptisine, palmatine, and berberine in regular sequence. RC-mix, a water extract of Rhei rhizoma and Coptis rhizoma mixture.
Figure 2
Figure 2
DPPH radical scavenging activity (a) and ABTS radical scavenging activity (b) of RC-mix, water extract of Rhei rhizoma and Coptis rhizoma mixture. Each experiment was run in triplicate.
Figure 3
Figure 3
Gross evaluation of the esophageal mucosal damage. (a) Representative microphotographs of the esophagus. Esophageal lesion observed in rats with induced reflux esophagitis (RE) was ameliorated by RC-mix (100, 200, and 400 mg/kg body weight/day, p.o.) administration. (b) Gross mucosal injury ratio at the end of experiment. The gross mucosal injury was increased in RE rats compared with normal rats, but RC-mix administration led to a significant decrease (RC200, p < 0.01; RC400, p < 0.001). N, normal rats; Veh, positive control rats with reflux esophagitis (RE); RC100, RC200, and RC400 RE rats, animals treated with RC-mix 100 mg/kg, RC-mix 200 mg/kg, and RC-mix 400 mg/kg body weight, respectively. Data are the means ± SEM. Significance: p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus RE control rat values. n = 6 in each group.
Figure 4
Figure 4
Effect of RC-mix on serum ROS and ONOO production in rats with induced reflux esophagitis (RE). N, normal rats; Veh, positive control rats with reflux esophagitis (RE); RC100, RC200, and RC400 RE rats, animals treated with RC-mix 100 mg/kg, RC-mix 200 mg/kg, and RC-mix 400 mg/kg body weight, respectively. Data are the means ± SEM. Significance: p < 0.05, ∗∗ p < 0.01 versus RE control rat values. n = 6 in each group.
Figure 5
Figure 5
Esophageal Nrf-2 (a), HO-1 (b), SOD (c), and catalase (d) protein expressions. N, normal rats; Veh, positive control rats with reflux esophagitis (RE); RC100, RC200, and RC400 RE rats, animals treated with RC-mix 100 mg/kg, RC-mix 200 mg/kg, and RC-mix 400 mg/kg body weight, respectively. Data are the means ± SEM. Significance: p < 0.05, ∗∗ p < 0.01 versus RE control rat values. n = 6 in each group.
Figure 6
Figure 6
Esophageal p-IκBα (a) and NF-κBp65 (b) protein expressions. N, normal rats; Veh, positive control rats with reflux esophagitis (RE); RC100, RC200, and RC400 RE rats, animals treated with RC-mix 100 mg/kg, RC-mix 200 mg/kg, and RC-mix 400 mg/kg body weight, respectively. Data are the means ± SEM. Significance: p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus RE control rat values. n = 6 in each group.
Figure 7
Figure 7
Esophageal COX-2 (a), iNOS (b), TNF-α (c), and IL-6 (d) protein expressions. N, normal rats; Veh, positive control rats with reflux esophagitis (RE); RC100, RC200, and RC400 RE rats, animals treated with RC-mix 100 mg/kg, RC-mix 200 mg/kg, and RC-mix 400 mg/kg body weight, respectively. Data are the means ± SEM. Significance: p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus RE control rat values. n = 6 in each group.
Figure 8
Figure 8
Predicted mechanism in esophageal tissue on administering RC400. RC400 decreased serum ROS and ONOO production. Further, RC400 ameliorated the values of proinflammatory mediators (COX-2 and iNOS) and cytokines (TNF-α and IL-6) regulated by NF-κB and increased oxidative defense factor SOD and HO-1 proteins.

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