Oxidative Medicine and Cellular Longevity / 2020 / Article / Fig 2

Research Article

Amarogentin from Gentiana rigescens Franch Exhibits Antiaging and Neuroprotective Effects through Antioxidative Stress

Figure 2

Effect of amarogentin on oxidative stress in yeast and the SOD, GPx, and CAT enzymatic activities of yeast. (a) The growth of yeast under oxidative stress induced by 10 mM H2O2 after treatment with amarogentin. The wild-type BY4741 yeast was inoculated in YPD medium for 24 h. Then after, initial 0.1 OD of yeast was placed in a liquid glucose medium and treated with RES at 10 μM as positive control or amarogentin (0, 1, 3, and 10 μM) for 24 h at 28°C. Subsequently, 5 μl of the cultured cells with the same OD600 value from each group was dropped on a plate containing 10 mM H2O2. The growth of yeast cells on the plate was observed and photographed after 3 days of incubation at 28°C. (b) Survival rate of the BY4741 yeast strain under 5.5 mM H2O2-induced yeast oxidative stress after adding amarogentin. BY4741 yeast cells were treated with RES (10 μM) or amarogentin (0, 1, 3, and 10 μM). The counted 200 yeast cells from each group were spread on a glucose agar plate with or without 5.5 mM H2O2 and incubated at 28°C for 48 h. After 2 days, the colonies that formed on the plate were counted. The survival rate of yeast cells was analysed from the ratio of the number of colonies in the absence of 5.5 mM H2O2 divided by the number of colonies in the presence of 5.5 mM H2O2. Effect of amarogentin on the total superoxide dismutase (T-SOD) (c), SOD1 (d), SOD2 (e), total glutathione peroxidase (GPx) (f) and catalase (CAT) (g). BY4741 yeast strain cells were incubated with RES as a positive control or amarogentin at concentrations of 1, 3, and 10 μM for 24 h. Afterwards, the yeast cells were collected by centrifugation and ultrasonicated for five times. The supernatant of the yeast was used to measure the T-SOD, SOD1, SOD2, GPx, and CAT activities according to the manufacturer’s instructions. Each experiment was conducted thrice. , , and indicated significant difference as compared with the control (, 0.01, and 0.001), respectively.

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