基于VEGF/p38MAPK研究羟基红花黄色素A对过氧化氢诱导人肝细胞氧化损伤的保护作用  

To investigate the protective effect of Hydroxysafflor yellow A on oxidative damage of hu⁃man hepatocytes induced by hydrogen peroxide based on VEGF/p38MAPK

作　　者：白皓天 杨婧[2] 李娅兰 梁霄 张筠昊 孙淑慧 张倩倩 王锐[1,3] BAI Hao-tian;YANG Jing;LI Ya-lan;LIANG Xiao;ZHANG Jun-hao;SUN Shu-hui;ZHANG Qian-qian;WANG Rui(College of Pharmacy,Heilongjiang University of Chinese Medicine,Harbin 150040,China;College of Basic Medical,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Key Laboratory of Basic and Application Research of Beiyao Heilongjiang University of Chinese Medicine,Ministry of Education,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学药学院,黑龙江哈尔滨150040 [2]黑龙江中医药大学基础医学院,黑龙江哈尔滨150040 [3]教育部北药基础与应用研究重点实验室,黑龙江哈尔滨150040

出　　处：《时珍国医国药》2023年第4期843-848,共6页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81603418);黑龙江省省属本科高校中央支持地方高校改革发展项目(2020YQ05);黑龙江中医药大学“优秀创新人才支持计划”科研项目(201208);黑龙江省“头雁”团队项目。

摘　　要：目的探讨羟基红花黄色素A(Hydroxysafflor yellow A,HSYA)对过氧化氢(H_(2)O_(2))诱导的人肝细胞LO2氧化损伤的保护作用及其机制。方法利用H_(2)O_(2)诱导并建立了LO2细胞氧化损伤模型。将细胞分为对照组(Control group,CG)、H_(2)O_(2)损伤模型组(Model group,MG)、HYSA(0.125,0.25,0.5,1mg·mL^(-1))干预组(MG+0.125,0.25,0.5,1mg·mL^(-1)HYSA)。分别利用MTT法、流式细胞术测定了细胞活力和凋亡程度;利用ELISA测定细胞内ROS、LDH、SOD、GSH-Px、MDA和CAT的活性及含量;Hoechst染色进一步观察细胞核凋亡状态;通过qRT-PCR技术测定细胞内VEGF、p38MAPK和凋亡相关蛋白的mRNA表达量;采用Western blotting法检测凋亡相关因子及VEGF/p38MAPK通路的蛋白表达。结果成功建立由H_(2)O_(2)所诱导的人肝细胞LO2损害模型;与MG比较,在0.125、0.25、0.5、1mg·mL^(-1)HYSA干预组细胞中LDH、MDA和ROS的活性和含量均明显下降,SOD、GSH-Px和CAT活性均明显增强,HYSA可明显抑制细胞凋亡并下调VEGF、p38MAPK/p-p38MAPK蛋白表达。结论HYSA可缓解H_(2)O_(2)对人肝LO2细胞造成的损伤,对细胞内氧化指标产生显著影响,有明显的氧化损伤保护作用,机制与VEGF/p38MAPK通路有关,可为HYSA的临床应用提供支持。Objective To investigate the protective effect of Hydroxysafflor yellow A(HSYA)on H_(2)O_(2)induced oxidative damage of human hepatocytes LO2 and its mechanism.Methods Oxidative damage Model of LO2 cell was established by H_(2)O_(2)induction.The cell were divided into Control group(CG)and Model group(MG),HYSA(0.125,0.25,0.5,1mg·ml^(-1))(MG+0.125,0.25,0.5,1mg·ml^(-1)HYSA).Cell viability and apoptosis were determined by MTT assay and flow cytometry respectively.The activities and contents of ROS,LDH,SOD,GSH-Px,MDA and CAT in cell were determined by ELISA.Hoechst staining was used to further observe the state of nuclear apoptosis.The mRNA expression levels of VEGF、p38MAPK and apoptosis-related proteins were determined by qRT-PCR.Western blotting was used to detect the expression of apoptosis-related factors and pro⁃tein of VEGF/p38MAPK pathway.Results The human hepatocyte LO2 damage model induced by H_(2)O_(2)was established success⁃fully.Compared with MG,the activities and contents of LDH,MDA and ROS in 0.125,0.25,0.5 and 1mg·ml^(-1)HYSA treat⁃ment groups were significantly decreased,while the activities of SOD,GSH-Px and CAT were significantly increased.HYSA could significantly inhibit cell apoptosis and down-regulate the protein expression of p38MAPK/p-p38MAPK.Conclusion⁃HYSA could relieve the damage of human hepatocytes LO2 caused by H_(2)O_(2),significantly affect the intracellular oxidation index,and have an obvious protective effect on oxidative damage.The mechanism is related to the VEGF/p38MAPK pathway,which can provide support for the clinical application of HYSA.

关 键 词：羟基红花黄色素A LO2细胞 氧化应激 P38MAPK VEGF 

分 类 号：R285.5[医药卫生—中药学]