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易鹏吉,张哲宇,彭伟军.槲皮素对H2O2诱导PC12细胞损伤的保护作用及机制研究[J].湖南中医药大学学报英文版,2021,41(6):821-826.[Click to copy
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| 槲皮素对H2O2诱导PC12细胞损伤的保护作用及机制研究 |
| 易鹏吉,张哲宇,彭伟军 |
| (中南大学湘雅二医院中西医结合科, 湖南 长沙 410011) |
| 摘要: |
| 目的 探讨槲皮素对H2O2损伤的大鼠嗜铬细胞瘤（PC12）细胞的保护作用及机制。方法 先选取不同浓度H2O2（100、200、300、400、500 μmol/L）干预PC12细胞，确定氧化损伤模型的条件，然后将PC12细胞分为对照组、模型组和槲皮素（6.25、12.5、25 μmol/L）组。通过MTS法、Hoechst染色法检测细胞存活和凋亡；镜下观察细胞形态；免疫荧光检测8-羟脱氧鸟苷（8-OHdG）；ELISA法检测丙二醛（MDA）和谷胱甘肽过氧化物酶（GSH）；Western blot法检测B淋巴细胞瘤2（Bcl-2）、Bcl-2相关X蛋白（Bax）、蛋白激酶B（Akt）和磷酸化蛋白激酶B（p-Akt）表达水平。结果 与对照组相比，模型组用500 μmol/L H2O2干预6 h后，细胞存活率下降至50%（P<0.01），凋亡率上升为38%（P<0.01）；镜下可见细胞萎缩变圆；8-OHdG表达增加（P<0.05）；GSH活性降低，MDA含量升高（P<0.05）；Bcl-2、Akt、p-Akt表达减少，Bax表达增加（P<0.05）。与模型组相比，槲皮素各浓度组细胞存活率均上升（P<0.05），凋亡率下降（P<0.01）；细胞突触再现；8-OHdG表达降低（P<0.05）；GSH活性升高，MDA含量下降（P<0.05）；Bcl-2、Akt、p-Akt蛋白上调，Bax蛋白下调（P<0.05）。结论 槲皮素能拮抗H2O2诱导的PC12细胞氧化损伤，其作用机制可能与Akt调控的通路以及抗氧化酶的相关作用有关。 |
| 关键词: 槲皮素 PC12细胞 氧化应激 凋亡 蛋白激酶B |
| DOI：10.3969/j.issn.1674-070X.2021.06.001 |
| Received:March 01, 2021 |
| 基金项目:国家自然科学基金面上项目（81873169）；湖南省自然科学基金面上项目（2020JJ4803）；湖南省中医药科研计划项目（201225）。 |
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| Protective Effect and Mechanism of Quercetin on H2O2-induced PC12 Cell Injury |
| YI Pengji,ZHANG Zheyu,PENG Weijun |
| (Department of Integrated Traditional Chinese and Western Medicine, The Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, China) |
| Abstract: |
| Objective To investigate the protective effect and its mechanisms of quercetin in rat pheochromocytoma (PC12 cell) cell injury stimulated by H2O2. Methods Different concentrations of H2O2 (100, 200, 300, 400, 500 μmol/L) were used to interfere with PC12 cells to construct oxidative stress model, then the experiments were divided into control group, model group and quercetin group (6.25, 12.5, 25 μmol/L). The cell survival and apoptosis were detected by MTS and Hoechst, respectively. Morphology was observed by microscope, and 8-hydroxydeoxyguanosine (8-OHdG) expression was measured by immunofluorescence. Furthermore, the levels of malondialdehyde (MDA) and glutathione (GSH) were assayed by ELISA and the expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), protein kinase B （Akt）and phosphorylated protein kinase B (p-Akt) were evaluated by Western blot. Results After constructed via 500 μmol/L H2O2 for 6 h, compared with the control group, cell survival rate in model group was decreased to 50% (P<0.01), apoptosis rate was increased to 38% (P<0.01); microscopically, the cells became round and atrophic; 8-OHdG expression increased (P<0.05); GSH activity was reduced, and the content of MDA was increased (P<0.05); Bcl-2, Akt and p-Akt expression levels were decreased, and Bax expression was increased (P<0.05). Compared with the model group, the cell survival rate of quercetin groups increased (P<0.05) and apoptosis rate decreased (P<0.01); cell synapse reappeared; the expression of 8-OHdG decreased (P<0.05); the level of GSH increased and the level of MDA decreased (P<0.05); the expression levels of Bcl-2, Akt and p-Akt were up-regulated, and the expression of Bax was down-regulated (P<0.05). Conclusion Quercetin could relieve oxidative damage of PC12 cells induced by H2O2. Its mechanism may be related to Akt regulated pathway and the related effects of antioxidant enzymes. |
| Key words: quercetin PC12 cell oxidative stress apoptosis Akt |
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