| 引用本文: | 苑舒文,董艺薇,刘健,梁亚杰,黄建军,肖保国,王青,马存根.原花青素B2对H2O2诱导的星形胶质细胞氧化损伤和凋亡的保护作用及机制研究[J].中国现代应用药学,2024,41(6):727-735. |
| YUAN Shuwen,DONG Yiwei,LIU Jian,LIANG Yajie,HUANG Jianjun,XIAO Baoguo,WANG Qing,MA Cungen.Protective Effect and Mechanism of Proanthocyanidin B2 Against H2O2-induced Oxidative Damage and Apoptosis of Astrocytes[J].Chin J Mod Appl Pharm(中国现代应用药学),2024,41(6):727-735. |
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| 原花青素B2对H2O2诱导的星形胶质细胞氧化损伤和凋亡的保护作用及机制研究 |
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苑舒文1, 董艺薇1, 刘健1, 梁亚杰1, 黄建军2, 肖保国3, 王青1, 马存根1
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1.山西中医药大学国家中医药管理局多发性硬化益气活血重点研究室，神经生物学研究中心，山西 晋中 030619;2.山西省卫健委神经疾病防治研究重点实验室/国药同煤总医院神经外科，山西 大同 037003;3.复旦大学附属华山医院神经内科，医学神经生物学国家重点实验室，上海 200025
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| 摘要: |
| 目的 探讨原花青素B2(proanthocyanidin B2，PC-B2)对过氧化氢(H2O2)诱导的小鼠星形胶质细胞(astrocytes，AS)氧化损伤和凋亡的保护作用及其机制。方法 利用C57BL/6新生小鼠(1~3 d)分离、培养AS，通过筛选的H2O2和PC-B2最佳作用浓度分为正常组、正常+PC-B2组(100 μg·mL-1 PC-B2处理24 h)、H2O2模型组(200 μmol·L-1 H2O2处理24 h)、PC-B2组(200 μmol·L-1 H2O2与100 μg·mL-1 PC-B2共同处理24 h)；CCK-8法检测各组细胞存活率，LDH法进行细胞毒性检测；ABTS和DPPH法检测PC-B2的抗氧化能力；ELISA试剂盒检测各组细胞中MDA含量以及SOD、CAT和GSH-Px活力；TUNEL染色法检测各组细胞凋亡情况；RT-PCR和Western blotting分别检测AS中Bax、Bcl-2、Caspase-3、Akt/Stat3、p-Akt、p-Stat3、Nrf2/HO-1的mRNA和蛋白表达水平。结果 PC-B2能够明显增强细胞活力，抑制AS凋亡。并且与H2O2模型组相比，PC-B2干预能够显著降低AS中LDH、MDA含量，提高SOD、CAT和GSH-Px活力，抑制Bax、Caspase-3的mRNA和蛋白表达，上调Akt/Stat3、Bcl-2、Nrf2/HO-1的mRNA和蛋白表达。结论 PC-B2能够通过Akt/Stat3和Nrf2/HO-1途径增强AS抗氧化能力，减轻H2O2诱导的AS氧化损伤和凋亡。 |
| 关键词: 原花青素B2 星形胶质细胞 H2O2 氧化损伤 细胞凋亡 Akt/Stat3 Nrf2/HO-1 |
| DOI：10.13748/j.cnki.issn1007-7693.20230642 |
| 分类号:R285.5 |
| 基金项目:国家自然科学基金青年项目(81903596)；山西省归国留学人员科研项目(2022-165)；山西中医药大学科技创新团队(2022TD2006)；山西中医药大学研究生创新创业项目(2021CX009，2022CX018) |
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| Protective Effect and Mechanism of Proanthocyanidin B2 Against H2O2-induced Oxidative Damage and Apoptosis of Astrocytes |
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YUAN Shuwen1, DONG Yiwei1, LIU Jian1, LIANG Yajie1, HUANG Jianjun2, XIAO Baoguo3, WANG Qing1, MA Cungen1
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1.The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine, Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, China;2.The Key Laboratory of Prevention and Treatment of Neurological Disease of Shanxi Provincial Health Commission/Department of Neurosurgery, Sinopharm Tongmei General Hospital, Datong 037003, China;3.State Key Laboratory of Neurobiology, Department of Neurology, Huashan Hospital, Fudan University, Shanghai 200025, China
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| Abstract: |
| OBJECTIVE To investigate the protective effect proanthocyanidin B2(PC-B2) on oxidative damage and apoptosis of mouse astrocytes(AS) induced by hydrogen peroxide(H2O2) and its mechanism. METHODS AS were isolated and cultured from neonatal C57BL/6 mice(1-3 d). The optimal concentration of H2O2 and PC-B2 was divided into four groups: normal group, normal+PC-B2 group(100 μg·mL-1 PC-B2 treated for 24 h), H2O2 model group(200 μmol·L-1 H2O2 treated for 24 h), PC-B2 group(200 μmol·L-1 H2O2 and 100 μg·mL-1 PC-B2 treated for 24 h). The cell viability of each group was detected by CCK-8 method. Cytotoxicity was detected by LDH method. The antioxidant capacity was detected by ABTS and DPPH. The content of MDA and the activity of SOD, CAT and GSH-Px were detected by ELISA kit. Detection of apoptosis in each group was done by TUNEL staining. The mRNA and protein expression levels of Bax, Bcl-2, Caspase-3, Akt/Stat3, p-Akt, p-Stat3 and Nrf2/HO-1 in AS were detected by RT-PCR and Western blotting, respectively. RESULTS PC-B2 could significantly enhance cell viability and inhibit AS apoptosis. Compared with the H2O2 model group, PC-B2 intervention could significantly reduce the content of LDH and MDA in AS, and increase the activity of SOD, CAT and GSH-Px. PC-B2 intervention could inhibit the mRNA and protein expression of Bax and Caspase-3, and up-regulate the mRNA and protein expression of Akt/Stat3, Bcl-2, Nrf2/HO-1. CONCLUSION PC-B2 can enhance the antioxidant capacity of AS through Akt/Stat3 and Nrf2/HO-1 pathways, therefore reduce H2O2-induced AS oxidative damage and apoptosis. |
| Key words: proanthocyanidin B2 astrocytes H2O2 oxidative damage cell apoptosis Akt/Stat3 Nrf2/HO-1 |
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