Abstract
BackgroundIn this study, Malus doumeri leaf flavonoids (MDLF) were used as the research object to observe their in vitro antioxidant stress ability. Hydrogen peroxide (H2O2) was used to induce oxidative stress in 293 T cells.MethodsMTT, flow cytometry, and qPCR were used to verify the effect of MDLF.ResultsIn vitro cell experiments showed that at a concentration of 0–160 μg/mL, MDLF did not affect the normal proliferation of human embryonic kidney 293 T cells (HEK 293 T cells), and MDLF had no cytotoxic effect in this concentration range. It was found that MDLF could maintain the survival of HEK 293 T cells (82.6%) at a high concentration (160 μg/mL). Morphological observation also found that MDLF can inhibit the cell structure imperfection caused by H2O2. It was also observed that MDLF could significantly increase the levels of catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), and glutathione peroxidase (GSH-Px) and reduce the level of malondialdehyde (MDA). The results of quantitative polymerase chain reaction (qPCR) showed that MDLF could significantly up-regulate the mRNA expression levels of CAT, SOD, GSH, GSH-Px, B-cell lymphoma-2 (Bcl-2) and downregulate the expression levels of B-cell lymphoma-2 associated x protein (Bax), tumor necrosis factor-alpha (TNF-α), and nuclear factor kappa-B (NF-κB) in oxidative stress-injured cells. The HPLC analysis showed that MDLF contained hyperin, isoquercetin, quercitrin, hesperidin, myricetin, baicalin and quercetin.ConclusionFrom the experimental results, it was observed that MDLF has a strong anti-oxidation ability in vitro, and it can interfere with the oxidative stress damage caused by H2O2 in 293 T cells. Therefore, MDLF is a type of natural substance with good anti-oxidant effect, and it has the potential to interfere with many diseases.
Highlights
In this study, Malus doumeri leaf flavonoids (MDLF) were used as the research object to observe their in vitro antioxidant stress ability
Toxic effect of MDLF in HEK 293 T cells When HEK 293 T cells were treated with MDLF at concentrations of 20, 40, 60, 80, 100, 120, 140, 160, 180, and 200 μg/mL, compared with HEK 293 T cells that were not treated with MDLF, MDLF at concentrations of 20– 160 μg/mL did not significantly affect the proliferation of HEK 293 T cells, and the survival rate of HEK 293 T cells in this concentration range was more than 95% (Fig. 1)
Effect of MDLF on the change in morphology induced by hydrogen peroxide (H2O2) in HEK 293 T cells After cell culture, the normal HEK 293 T cells were found to be abundant in quantity, complete in cell structure, abundant in cytoplasm and orderly in cell arrangement in a microscopic field (Fig. 3); after being treated with H2O2, the number of HEK 293 T cells was decreased compared with the number of cells without H2O2 treatment
Summary
Malus doumeri leaf flavonoids (MDLF) were used as the research object to observe their in vitro antioxidant stress ability. Li et al BMC Complementary Medicine and Therapies (2020) 20:276 intracellular Ca2+ is increased under oxidative stress. It increases the expression of the apoptotic gene, but it decreases the expression of the anti-apoptotic protein, leading to normal cell apoptosis and pathological changes in the body [3]. Pathogens invade the body, activate neutrophils, increase oxygen consumption, and generate a large number of oxygen free radicals. At the same time, when pathogens invade and cause a stress reaction, they can promote the release of catecholamines and produce a large number of oxygen free radicals. The resulting oxidative stress reaction can lead to inflammatory infiltration of neutrophils and can produce a large number of pro-inflammatory mediators. Oxidative stress and inflammatory reaction can cause a variety of diseases, such as hypertension and atherosclerosis [5]
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