勒哲多糖粗提物体外抑制H_(2)O_(2)诱导HepG2细胞氧化损伤的作用及机制研究  

The effect of Lezhe polysaccharide crude extract on inhibiting H_(2)O_(2)-induced oxidative damage in HepG2 cells in vitro and its mechanism

作　　者：樊慧芳 李国峰 陈海芳 张武岗[1] FAN Hui-fang;LI Guo-feng;CHEN Hai-fang;ZHANG Wu-gang(Engineering Center for Solid Preparation in Chinese Herbal Medicine,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,China;Key Laboratory of Modern Preparation of Traditional Chinese Medicine,Ministry of Education,Jiangxi University of Traditional Chinese Medicine National Pharmaceutical,Nanchang 330004,China)

机构地区：[1]江西中医药大学中药固体制剂制造技术国家工程研究中心,南昌330004 [2]江西中医药大学现代中药制剂教育部重点实验室,南昌330004

出　　处：《中国新药杂志》2024年第8期810-817,共8页Chinese Journal of New Drugs

基　　金：国家自然科学基金资助项目(81903917);江西省教育厅资助项目(GJJ2200908);江西省自然科学基金资助项目(20192-BAB205109);博士科研启动基金资助项目(2018WBZR002)。

摘　　要：目的:基于体外活性氧(reactive oxygen species,ROS)清除体系和H_(2)O_(2)诱导HepG2细胞氧化损伤模型探讨勒哲多糖粗提物(LZC)的体外抗氧化作用和机制。方法:测定LZC对羟基自由基、超氧阴离子、DPPH自由基、亚硝酸根离子的清除作用;试剂盒检测H_(2)O_(2)诱导HepG2细胞氧化损伤模型中细胞中丙二醛(malondialdehyde,MDA)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平,2’,7’-二氯荧光黄双乙酸盐(2’,7’-dichlorofluorescein diacetate,DCFH2-DA)荧光探针检测细胞内ROS水平和Annexin V-FITC/PI双染法检测细胞凋亡情况,Western blot检测核因子E2相关因子2/血红素氧合酶-1(nuclear factor erythroid-2-related factor 2/heme oxygenase 1,Nrf2/HO-1)、肿瘤蛋白53(p53)、Bcl-2相关X蛋白(Bax)和半胱氨酸天冬氨酸蛋白水解酶-3(Caspase-3)的蛋白表达。结果:LZC表现出强ROS清除活性,显著提升H_(2)O_(2)诱导HepG2细胞氧化损伤模型中细胞的GSH-Px活性,抑制MDA水平、ROS释放和细胞凋亡。Western blot结果表明LZC能上调Nrf2和HO-1的蛋白表达,下调p53,Bax和Caspase-3表达。结论:LZC具有较强的体外抗氧化作用,其作用与激活Nrf2/HO-1信号通路进而抑制细胞凋亡有关。Objective:To investigate the in vitro antioxidant effect and mechanism of LZC based on ROS scavenging system and H_(2)O_(2)-induced oxidative damage model in HepG2 cells.Methods:The scavenging effect of LZC on hydroxyl radical,superoxide anion,DPPH radical and nitrite ion were determined.The levels of MDA and GSH-Px in H_(2)O_(2)-induced oxidative damage model of HepG2 cells were detected by kits.The intracellular ROS levels were detected by DCFH2-DA fluorescent probe.The apoptosis was detected by Annexin V-FITC/PI double staining.The protein expression levels of Nrf2/HO-1,p53,Bax and Caspase-3 were detected by Western blot.Results:LZC showed strong ROS scavenging activity,significantly increased GSH-Px activity in H_(2)O_(2)-induced oxidative damaged HepG2 cell model,and inhibited MDA level,ROS release and apoptosis.Western blot showed that LZC up-regulated the protein expressions of Nrf2 and HO-1,and downregulated the expressions of p53,Bax and Caspase-3.Conclusion:LZC has a strong antioxidant effect in vitro,which is related to the activation of Nrf2/HO-1 signaling pathway and the inhibition of apoptosis.

关 键 词：勒哲多糖粗提物 抗氧化 核因子E2相关因子2/血红素氧合酶-1信号通路 细胞凋亡 

分 类 号：R965[医药卫生—药理学]