| 摘要: |
| 目的:研究黄芪多糖(APS)在肝癌铁死亡机制的调控作用。方法:取不同浓度APS(0、50、100、200 mg/L)处理肝癌HepG2细胞48 h，分别记作对照组和APS低、中、高剂量组。采用CCK-8试剂盒检测细胞增殖情况，同时采用试剂盒检测各组细胞的谷胱甘肽(GSH)、活性氧自由基(ROS)和脂质过氧化物水平，采用流式细胞术检测各组细胞的铁离子水平。通过蛋白质印迹法检测各组细胞的铁死亡标志蛋白谷胱甘肽过氧化物酶4(GPX4)、长链脂酰CoA合成酶4(ACSL)以及Wnt/β-catenin信号通路关键蛋白Wnt和β-catenin蛋白的表达水平。结果:与对照组相比，APS高、中、低剂量组均能抑制肝癌HepG2细胞的增殖，且随着APS的浓度升高，抑制效果更显著(P<0.05)。与对照组相比，高、中、低剂量组均能降低肝癌HepG2细胞中GSH水平，并提高ROS和脂质过氧化物水平，同时提高铁离子水平，且随着APS浓度升高效果更显著(P<0.05)。同时，与对照组相比，APS高、中、低剂量组均能降低GPX4蛋白表达水平，提高ACSL4蛋白表达水平，且呈剂量±赖性(P<0.05)。与对照组相比，APS高、中、低剂量组均能降低Wnt和β-catenin蛋白表达水平，随着APS剂量升高，蛋白表达抑制效果更显著(P<0.05)。结论:APS通过增强ROS和脂质过氧化物的积累增强铁死亡，抑制肝癌细胞的增殖，这一过程可能与Wnt/β-catenin信号通路的抑制相关。 |
| 关键词: 黄芪多糖 铁死亡 肝癌 Wnt/β-catenin信号通路 增殖 |
| DOI：10.3969/j.issn.1007-6948.2025.01.023 |
| 投稿时间：2024-07-08 |
| 基金项目: |
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| Astragalus polysaccharide promotes ferroptosis and inhibits proliferation of hepatocellular carcinoma cells by regulating Wnt/β-catenin signaling pathway |
| ZHU Li-qing,WANG Zhi-xin,CAO Hu |
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| Abstract: |
| Objective To investigate the regulatory effect of astragalus polysaccharide (APS) on ferroptosis mechanism of hepatocellular carcinoma. Methods HepG2 cells were treated with different concentrations of APS (0, 50, 100, 200 mg/L) for 48 hours, and were recorded as control group and APS low, medium and high dose group, respectively. Cell proliferation was detected with CCK-8 kit, while glutathione (GSH), Reactive Oxygen Species (ROS) and lipid peroxide levels of each group were detected with commercial kits, respectively. Flow cytometry was used to detect the iron level in each group. Glutathione Peroxidase 4 (GPX4) and long-chain acyl-CoA synthetase 4 (ACSL4), markers of ferroptosis, were detected by Western blot, as well as the expression levels of key proteins of the Wnt/β-catenin signaling pathway including Wnt and β-catenin. Results Compared with the control group, APS high-dose, medium-dose and low-dose groups could inhibit the proliferation of HepG2 cells, and the inhibitory effect was more significant with the increase of APS concentration (P<0.05). In addition, compared with the control group, the high, medium and low dose groups could reduce the GSH level in HepG2 cells, increase the ROS and lipid peroxide levels, and increase the iron level, and the effect was more significant with the increase of APS concentration (P<0.05). At the same time, Western blot further verified that compared with the control group, APS high, medium and low dose groups could reduce GPX4 protein expression level and increase ACSL4 protein expression level in a dose-dependent manner (P<0.05). Compared with the control group, the expression levels of Wnt and β-catenin protein in high, medium and low dose APS groups were decreased, and the inhibitory effect of protein expression was more significant with the increase of APS dose (P<0.05). Conclusion APS can enhance the accumulation of ROS and lipid peroxides, enhance ferroptosis and inhibit the proliferation of hepatocellular carcinoma cells, which may be related to the deactivation of Wnt/β-catenin signaling pathway. |
| Key words: Astragalus polysaccharide ferroptosis hepatocellular carcinoma Wnt/β-catenin signal pathway proliferation |
