Molecular Bioprospection Department of Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow 226015, Uttar Pradesh, India.
Molecular Bioprospection Department of Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow 226015, Uttar Pradesh, India.
Phytomedicine. 2019 Mar 1;55:92-104. doi: 10.1016/j.phymed.2018.07.009. Epub 2018 Jul 20.
Rutin (quercetin-3-O-rutinoside), a flavonoid, is predominantly found in the buckwheat, cranberries, mulberry and citrus fruits. It is used as a restorative in the preparation of herbal medicine, multivitamin and known to reduce the fate of heart attack.
We aimed to elucidate whether rutin attenuates oxidative stress and its possible mechanism of action in ameliorating the deleterious effect of t-BHP. We also provide evidence that rutin protects the antioxidant status of erythrocytes and liver via Nrf2 and iNOS pathway from oxidative stress.
STUDY DESIGN/METHOD: Human erythrocytes and mice liver were used for the evaluation of rutin's effect against t-BHP induced oxidative stress. The non-enzymatic (GSH, MDA, -CO, -SH) and enzymatic stress markers (SOD, CAT, GPx, GR and GST) were estimated by the colorimetric method. The level of Nrf2, iNOS, liver marker enzymes, triglycerides, cholesterol, HDL-cholesterol, albumin, BUN was measured using ELISA kits. Reactive oxygen species (ROS) was quantified using flow cytometry and fluorometry. RT-PCR was used for the quantification of Nrf2 and iNOS expression levels in the liver tissue of mice. In silico studies were done through receptor-ligand binding interaction.
Pre-treatment with the rutin ameliorated the toxic effect of t-BHP by modulating the basal level of GSH, -SH, MDA and -CO significantly (p < 0.01) with respect to untreated control. Rutin also protected the erythrocytes against the t-BHP-induced oxidative stress as evidenced by augmented activity of antioxidant enzymes (CAT, SOD, GP, GR and GST). Furthermore, at the highest tested concentration (16.3 µM), it protected the morphology of the erythrocytes by decreasing the ROS level (p < 0.01). In addition, the lower MEF values of rutin (0.520 ± 0.005) alone or along with t-BHP (0.630 ± 0.021) indicated its non-toxic and protective behavior. The qPCR analyses revealed that t-BHP potently up-regulates the iNOS and down regulate the Nrf2 expression which was ameliorated with rutin treatment in a dose-dependent manner like silymarin.
Our findings demonstrate that rutin potentiates its beneficial aspect by displaying a profound role in iNOS-Nrf2 signaling pathway. Accordingly, it may be concluded that the dietary factors wherein rutin is an ingredient could be helpful in the maintenance of the intracellular redox-homeostasis and thus may be effective against oxidative stress related secondary complications.
芦丁（槲皮素-3-O-芸香糖苷）是一种黄酮类化合物，主要存在于荞麦、蔓越莓、桑椹和柑橘类水果中。它被用作草药、多种维生素制剂中的滋补品，已知可降低心脏病发作的几率。
我们旨在阐明芦丁是否通过 Nrf2 和 iNOS 途径减轻 t-BHP 的有害作用来减轻氧化应激，并提供芦丁通过抗氧化应激保护红细胞和肝脏的抗氧化状态的证据。
研究设计/方法：用人红细胞和小鼠肝评估芦丁对 t-BHP 诱导的氧化应激的作用。通过比色法测定非酶（GSH、MDA、-CO、-SH）和酶应激标志物（SOD、CAT、GPx、GR 和 GST）。使用 ELISA 试剂盒测量 Nrf2、iNOS、肝标志物酶、甘油三酯、胆固醇、HDL-胆固醇、白蛋白、BUN 的水平。使用流式细胞术和荧光法定量活性氧（ROS）。通过 RT-PCR 定量小鼠肝组织中 Nrf2 和 iNOS 的表达水平。通过受体-配体结合相互作用进行计算机模拟研究。
芦丁预处理通过显著调节 GSH、-SH、MDA 和 -CO 的基础水平来改善 t-BHP 的毒性作用（p < 0.01）与未处理的对照相比。芦丁还通过增强抗氧化酶（CAT、SOD、GP、GR 和 GST）的活性来保护红细胞免受 t-BHP 诱导的氧化应激。此外，在最高测试浓度（16.3 µM）下，它通过降低 ROS 水平来保护红细胞的形态（p < 0.01）。此外，芦丁的较低 MEF 值（0.520 ± 0.005）单独或与 t-BHP 一起（0.630 ± 0.021）表明其无毒且具有保护作用。qPCR 分析表明，t-BHP 强力上调 iNOS 并下调 Nrf2 表达，芦丁处理以剂量依赖性方式改善这种情况，类似于水飞蓟素。
我们的研究结果表明，芦丁通过在 iNOS-Nrf2 信号通路中发挥重要作用，增强其有益方面。因此，可以得出结论，饮食因素中含有芦丁可能有助于维持细胞内氧化还原稳态，从而可能有效对抗与氧化应激相关的继发性并发症。
