首页|勒哲多糖粗提物体外抑制H2O2诱导HepG2细胞氧化损伤的作用及机制研究

勒哲多糖粗提物体外抑制H2O2诱导HepG2细胞氧化损伤的作用及机制研究

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目的:基于体外活性氧(reactive oxygen species,ROS)清除体系和H2 O2 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H2 O2 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H2 O2 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.
The effect of Lezhe polysaccharide crude extract on inhibiting H2O2-induced oxidative damage in HepG2 cells in vitro and its mechanism
Objective:To investigate the in vitro antioxidant effect and mechanism of LZC based on ROS scavenging system and H2 O2-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 H2 O2-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 H2 O2-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.

Lezhe polysaccharide crude extractantioxidationnuclear factor erythroid-2-related factor 2/heme oxygenase-1 signaling pathwaycell apoptosis

樊慧芳、李国峰、陈海芳、张武岗

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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

江西中医药大学现代中药制剂教育部重点实验室,南昌 330004

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

国家自然科学基金江西省教育厅项目江西省自然科学基金博士科研启动基金

81903917GJJ220090820192-BAB2051092018WBZR002

2024

中国新药杂志
中国医药科技出版社 中国医药集团总公司 中国药学会

中国新药杂志

CSTPCD北大核心
影响因子：1.039
ISSN：1003-3734
年,卷(期)：2024.33(8)
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