Antioxidant Activity of Tea Polysaccharide and Its Protective Mechanism Against Oxidative Damage

doi:10.11983/CBB22088

Abstract

Abstract:

Wan tea polysaccharides (WTPs) is a kind of acid glycoprotein extracted from tea, which has good antioxidant activity. The antioxidant activity of WTPs in Southwest Anhui was analyzed using the scavenging rate of free radicals as an index, and the repair effect of WTPs on free radical damage was further explored based on the exogenous ·OH injury cell model established by H₂O₂ and EDTA-Fe²⁺ and the endogenous hydroxyl radical injury model induced by PMA. The results showed that WTPs had good antioxidant activity against DPPH· and ·OH, with EC₅₀ values of 209.5 and 535.2 µg∙mL^-1 on scavenging efficiency, respectively, which was similar with Vc. The survival rate of cells in both exogenous and endogenous free radical oxidative damage models increased with the increase of WTPs concentration by cell proliferation assay, which could reach 87.41% and 85.84% when WTPs concentration was 800 µg∙mL^-1, which was significantly higher than that in model group (47.67% and 48.03%). In terms of protective mechanism, laser confocal imaging of reactive oxygen species (ROS) distribution in cells and fluorescence intensity analysis showed that compared with the model group, WTPs had significant scavenging effect on exogenous and endogenous ROS in cell models, which was consistent with the results of antioxidant test in vitro. WTPs showed good free radical scavenging efficiency in vitro and its improvement effect on free radical damage at the cellular level. The study revealed antioxidant damage mechanism of WTPs in scavenging free radicals at the cellular level, which laid a foundation for further research on the anti-aging effect of tea polysaccharides on the body aging caused by free radicals.

Key words: tea polysaccharides, antioxidant, cell oxidative damage, repair mechanism

Zhang Mingzhu, Qin Huaguang, Mu Dan, Yang Longyu, Li Hu, Qi Zehua, Wang Yuxi, Zhang Yongcheng, Ye Lili, Yin Wenjing, Wang Shuyuan, Rao Yuchun, Wu Yan. Antioxidant Activity of Tea Polysaccharide and Its Protective Mechanism Against Oxidative Damage[J]. Chinese Bulletin of Botany, 2022, 57(4): 444-456.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB22088

https://www.chinbullbotany.com/EN/Y2022/V57/I4/444

Figures/Tables 8

Figure 1 Free radical scavenging ability of different concentrations of Wan tea polysaccharides (WTPs) (A) DPPH·; (B)·OH; (C) O2-

Figure 2 Effects of Wan tea polysaccharides (WTPs) concentration on the survival rate of HepG-2 cells (A) and RAW264.7 cells (B)

Figure 3 Effects of Wan tea polysaccharides (WTPs) on cell physiological state (A) Morphology of mitochondria in control cells; (B) Zoom in micrograph from selected regions from different treated cells and their intensity analysis; (C) Morphology of mitochondria in WTPs incubated cells; (D) Zoom in micrograph from selected regions from different treated cells and their intensity analysis. DIC: Differential interference contrast. Bars=20 μm

Figure 4 Protective effect of Wan tea polysaccharides (WTPs) on demaged cells (A) Effects of different concentrations of exogenous ·OH on the survival rate of HepG-2 cells; (B) Effects of different concentrations of WTPs on HepG-2 cell survival rate induced by ·OH. Compared with control group, *P<0.05, **P<0.01; compared with model group, #P<0.05, ##P<0.01

Figure 5 Effect of Wan tea polysaccharides (WTPs) on the viability of RAW264.7 cells induced by PMA (A) Effects of endogenous ·OH induced by different concentration of PMA on the survival rate of RAW264.7 cells; (B) The effect of different concentration of PMA on morphological changes of RAW264.7 cells (bars=20 μm); (C) Effects of different concentration of WTPs on RAW264.7 cells survival rate induced by endogenous ·OH. Compared with control group, *P<0.05, **P<0.01; compared with model group, #P<0.05, ##P<0.01

Table 1 Effects of Wan tea polysaccharides (WTPs) on the content of glutathione reductase (GSH-PX) and catalase (CAT) in oxidation-damaged RAW264.7 cells (means±SD)

Group	GSH-PX	CAT
Group	(U∙mL^-1)	(U∙mL^-1)
Control	0.45±0.08	1.08±0.12
Model	0.13±0.04**	0.59±0.04**
100 µg∙mL^-1 WTPs	0.16±0.04**	0.57±0.06**
200 µg∙mL^-1 WTPs	0.15±0.04**	0.60±0.07**
400 µg∙mL^-1 WTPs	0.12±0.03**	0.62±0.05**
800 µg∙mL^-1 WTPs	0.13±0.02**	0.57±0.08**

Figure 6 Repair effect of Wan tea polysaccharides (WTPs) on oxidative damage of HepG-2 cells (A) Confocal imaging of ·OH content in cells of different treatment groups (bars=20 μm); (B) Comparison of fluorescence intensity of ·OH in cells of different treatment groups. Compared with control group, *P<0.05, **P<0.01; compared with model group, #P<0.05, ##P<0.01.

Figure 7 Effect of Wan tea polysaccharides (WTPs) on PMA-induced oxidative damage of RAW264.7 macrophages (A) Confocal imaging of ·OH content in cells of different treatment groups (bars=20 μm); (B) Comparison of fluorescence intensity of ·OH in cells of different treatment groups. Compared with control group, *P<0.05, **P<0.01; compared with model group, #P<0.05, ##P<0.01

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