7-羟乙基白杨素对PC12细胞的抗氧化作用及机制研究  

Study on Antioxidative Capacity and Mechanism of 7-Hydroxyethyl Chrysin in PC12 Cells

作　　者：刘菁 冷非凡[2] 曹琪璐 高玉海 马慧萍 陈克明 LIU Jing;LENG Feifan;CAO Qilu;GAO Yuhai;MA Huiping;CHEN Keming(Fundamental Medical Science Research Laboratories,The 940th Hospital of Joint Logistics Support Force of CPLA,Lanzhou 730050,China;School of Life Science and Engineering,Lanzhou University of Technology,Lanzhou 730050,China)

机构地区：[1]中国人民解放军联勤保障部队第九四〇医院基础医学实验室,兰州730050 [2]兰州理工大学生命科学与工程学院,兰州730050

出　　处：《中国现代应用药学》2021年第13期1537-1542,共6页Chinese Journal of Modern Applied Pharmacy

基　　金：国家自然科学基金项目(81571847)。

摘　　要：目的研究7-羟乙基白杨素对PC12细胞的抗氧化作用,并对其保护机制进行探讨。方法使用CCK-8试剂盒检测PC12细胞的存活率,筛选出7-羟乙基白杨素作用的最佳浓度进行实验。之后将PC12细胞随机分为4组,分别为对照组、缺氧组、白杨素组和7-羟乙基白杨素组。使用微量酶标法测定细胞培养基中LDH活性,DCFH-DA染色观察细胞内ROS含量,同时使用试剂盒对细胞内MDA、SOD和CAT水平进行测定,提取细胞总蛋白通过Western blotting检测评价Nrf2及其下游蛋白表达量。结果缺氧后PC12细胞存活率显著下降,经7-羟乙基白杨素预处理后可获得改善;白杨素组和7-羟乙基白杨素组上清液中的LDH和细胞内ROS、MDA含量与缺氧组相比显著降低,SOD和CAT含量与缺氧组相比显著升高,同时发现7-羟乙基白杨素在各个方面的保护作用都明显强于白杨素。缺氧条件会使Nrf2、Keap1、HO-1、NQO1蛋白表达增加,7-羟乙基白杨素干预后可以进一步提高它们的蛋白表达。结论 7-羟乙基白杨素具有比白杨素更好的保护效果,它可以通过激活Nrf2/ARE通路减轻PC12细胞由于缺氧造成的损伤。OBJECTIVE To explore the antioxidant effect and mechanism of 7-hydroxyethyl chrysin in PC12 cells. METHODS Cell viability was detected by CCK-8, and the best concentration of 7-hydroxyethyl chrysin was selected for subsequent experiments. Then PC12 cells were divided into 4 groups: control group, hypoxia group, chrysin group and 7-hydroxyethyl chrysin group. LDH in cell culture medium was evaluated by the microplate method. ROS level was evaluated by DCFH-DA. The levels of MDA, SOD and CAT in the cells were measured by the test kit. Western blotting was used to evaluate Nrf2 and its downstream protein expression. RESULTS The cell viability was reduced after hypoxia, it can be improved after pretreatment with 7-hydroxyethyl chrysin. LDH in the cell culture medium, ROS and MDA levels in the cells of chrysin group and 7-hydroxyethyl chrysin group were also significantly reduced compared with the hypoxia group, the activity of SOD and CAT was also significantly increased when compared with the hypoxia group, it was also found that the effect of 7-hydroxyethyl chrysin was significantly higher than chrysin in all aspects. Meanwhile, hypoxia could increase the protein expression of Nrf2, Keap1, HO-1, NQO1, and 7-hydroxyethyl chrysin could further improve their protein expression. CONCLUSION The protective effect of 7-hydroxyethyl chrysin is better than that of chrysin. It can reduce the damage of PC12 cells caused by hypoxia by activating the Nrf2/ARE pathway.

关 键 词：7-羟乙基白杨素 白杨素 PC12细胞 缺氧 Nrf2/ARE通路 

分 类 号：R965.1[医药卫生—药理学]