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引用本文:景临林,杨颖,武柠子,马慧萍,贾正平.黄芩素-7-甲醚对缺氧致PC12细胞损伤的保护作用[J].中国现代应用药学,2018,35(6):787-792.
JING Linlin,Yang Ying,WU Ningzi,MA Huiping,JIA Zhengping.Protective Effect of Negletein Against Hypoxia Induced Injury on PC12 Cells[J].Chin J Mod Appl Pharm(中国现代应用药学),2018,35(6):787-792.
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黄芩素-7-甲醚对缺氧致PC12细胞损伤的保护作用
景临林, 杨颖, 武柠子, 马慧萍, 贾正平
兰州总医院药剂科, 全军高原环境损伤防治重点实验室, 兰州 730050
摘要:
目的 研究黄芩素-7-甲醚对缺氧PC12细胞的保护作用。方法 将PC12细胞分为正常对照组、缺氧模型组、芦丁组和黄芩素-7-甲醚组,培养24 h后,MTT法检测黄芩素-7-甲醚对细胞活力的影响;酶标仪法检测细胞培养上清液中乳酸脱氢酶(lactate dehydrogenase,LDH)活性以及细胞内丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)水平;2',7'-二氯荧光素探针法检测细胞内活性氧簇(reactive oxygen species,ROS)水平;实时荧光定量PCR和Western blot检测核因子E2相关因子2(Nrf2)和血红素氧合酶-1(heme oxygenase-1,HO-1)的mRNA和蛋白的表达。结果 与正常对照组相比,缺氧导致PC12细胞活力显著降低,LDH、MDA和ROS水平显著升高,抗氧化酶SOD和CAT的活力显著降低。黄芩素-7-甲醚预处理能够逆转这些变化。此外,缺氧能够诱导细胞内Nrf2、HO-1的mRNA和蛋白表达增加,而黄芩素-7-甲醚能够进一步提高Nrf2、HO-1的mRNA和蛋白表达。结论 黄芩素-7-甲醚对缺氧致PC12细胞损伤有一定的保护作用,其作用机制可能与激活Nrf2/HO-1通路,抑制氧化应激损伤有关。
关键词:  黄芩素-7-甲醚  缺氧  PC12细胞  氧化应激  活性氧  Nrf2/HO-1通路
DOI:10.13748/j.cnki.issn1007-7693.2018.06.001
分类号:R285.5
基金项目:国家自然科学基金项目(81202458);全军医药卫生科研基金(CLZ12JA04);中国博士后科学基金(2012M521926);甘肃省省青年科技基金计划项目(1308RJYA061)
Protective Effect of Negletein Against Hypoxia Induced Injury on PC12 Cells
JING Linlin, Yang Ying, WU Ningzi, MA Huiping, JIA Zhengping
Department of Pharmacy, Lanzhou General Hospital, Key Lab of PLA for Prevention and Treatment of Injuries Induced by High Altitude, Lanzhou 730050, China
Abstract:
OBJECTIVE to investigate the protective effect of negletein against hypoxia induced injury on PC12 cells. METHODS The PC12 cells were divided into four groups, including normal control group, hypoxia model group, rutin group and negletein group. Cell viability was detected using MTT assay. Lactate dehydrogenase (LDH), malondialdehyde (MDA), catalase (CAT) and superoxide dismutase (SOD) in cell culture supernatant were evaluated by enzyme labeling method. Reactive oxygen species (ROS) level was evaluated by DCFH-DA. The expression of mRNA and protein of Nrf2 and heme oxygenase-1(HO-1) were assessed with RT-PCR analysis and Western blot. RESULTS Compared with normal control group, the viability as well as the activity of antioxidant enzyme such as SOD and CAT were reduced and the level of LDH, MDA and ROS were increased treated with hypoxia in PC12 cells. These changes were reversed with the pretreatment of negletein. Moreover, the expression of Nrf2 and HO-1 mRNA and proteins were also increased induced by hypoxia, even further upregulated by negletein pretreatment. CONCLUSION The results indicate that negletein can suppress the hypoxia-induced damage on PC12 cells by activating of the Nrf2/HO-1 pathway and alleviation of oxidative stress.
Key words:  negletein  hypoxia  PC12 cell  oxidant stress  reactive oxygen species  Nrf2/HO-1 pathway
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