[Antioxidant activity of Euryale ferox seed shell extract and its therapeutic effects on oral ulcer in rats].

Q Wang, F Xu, M Deng, M Ren, T Wang, D Wu
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Abstract

Objective: To investigate the therapeutic effect of Euryale ferox seed shell extract on oral ulcer in rats and its underlying mechanism.

Methods: The contents of polyphenols and flavonoids in Euryale ferox seed shells were determined by Folin-phenol assay and aluminum nitrate colorimetry, respectively. DPPH·, ABTS+·, ·OH and·O2- scavenging experiments were performed to evaluate the antioxidant activities of Euryale ferox seed shell extract in vitro. In a rat model of oral ulcer induced by burning with glacial acetic acid, the therapeutic effect of Euryale ferox seed shell extract was assessed by detecting changes in serum levels of oxidative factors by enzyme-linked immunosorbent assay (ELISA) and observing pathological changes of the ulcerous mucosa using HE staining; the therapeutic mechanism of the extract was explored by detecting the expression levels of Keap1, Nrf2, Nes-Nrf2 and HO-1 proteins in ulcerous mucosa using Western blotting.

Results: The ethyl acetate extract of Euryale ferox seed shells contained 306.74±1.04 mg/g polyphenols and 23.43±0.61 mg/g flavonoids and had IC50 values for scavenging DPPH· and ABTS+· free radicals of 3.42 ± 0.97 μg/mL and 3.32 ± 0.90 μg/mL, respectively. In the rat models, the ethyl acetate extract significantly ameliorated oral mucosal ulcer, increased serum CAT level, and decreased serum MDA level. The protein expression levels of Nes-Nrf2 and HO-1 were increased and Keap1 protein expression was lowered significantly in the ulcerous mucosa of the rats after treatment with the extract (P<0.05 or 0.01).

Conclusion: The therapeutic effect of Euryale ferox seed shell extract on oral ulcers in rats is mediated probably by activation of the Keap1/Nrf2/HO-1 signaling pathway.

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[阿魏籽壳提取物的抗氧化活性及其对大鼠口腔溃疡的治疗效果]。
目的方法:采用福林酚测定法和硝酸铝比色法分别测定阿魏多酚和黄酮类化合物的含量:方法:采用Folin-酚测定法和硝酸铝比色法分别测定白花蛇舌草种壳中多酚和黄酮类化合物的含量。进行了 DPPH-、ABTS+-、-OH 和-O2-清除实验,以评估欧亚种壳提取物的体外抗氧化活性。在冰醋酸灼烧诱导的大鼠口腔溃疡模型中,通过酶联免疫吸附试验(ELISA)检测血清中氧化因子水平的变化,并利用 HE 染色观察溃疡粘膜的病理变化,评估了白花蛇舌草种壳提取物的治疗效果;利用 Western 印迹技术检测了 Keap1、Nrf2、Nes-Nrf2 和 HO-1 蛋白在溃疡粘膜中的表达水平,探讨了白花蛇舌草种壳提取物的治疗机制:铁皮欧亚种壳乙酸乙酯提取物含有 306.74±1.04 mg/g 多酚和 23.43±0.61 mg/g 类黄酮,清除 DPPH 和 ABTS+ 自由基的 IC50 值分别为 3.42 ± 0.97 μg/mL 和 3.32 ± 0.90 μg/mL。在大鼠模型中,乙酸乙酯提取物能明显改善口腔黏膜溃疡,提高血清CAT水平,降低血清MDA水平。经该提取物治疗后,大鼠溃疡粘膜中 Nes-Nrf2 和 HO-1 蛋白表达水平明显提高,Keap1 蛋白表达水平明显降低:铁皮欧亚种壳提取物对大鼠口腔溃疡的治疗作用可能是通过激活 Keap1/Nrf2/HO-1 信号通路介导的。
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