圣草酚通过调节Nrf2通路促进氧化应激诱导的肝癌细胞的凋亡  

Eriodictyol promotes oxidative stress-induced apoptosis of liver cancer cells by regulating Nrf2 pathway

作　　者：张昆鹏[1] 张晓愉[1] 李少一[1] 甄品 尤江莲[1] ZHANG Kun-Peng;ZHANG Xiao-Yu;LI Shao-Yi;ZHEN Pin;YOU Jiang-Lian(Xingtai People's Hospital,Xing-tai 054001,China)

机构地区：[1]邢台市人民医院,邢台054001

出　　处：《中国免疫学杂志》2021年第12期1449-1453,1467,共6页Chinese Journal of Immunology

基　　金：河北省重点研发计划项目(172777214)资助。

摘　　要：目的:探究圣草酚对氧化应激诱导的肝癌细胞凋亡的作用机制。方法:CCK-8测定圣草酚对细胞的毒性。将Huh7细胞分为对照组、H_(2)O_(2)组、圣草酚组、H_(2)O_(2)+圣草酚组,分别用0.5 mmol/L H_(2)O_(2)或78μmol/L圣草酚处理细胞48 h;2,7-二氯荧光素双乙酸酯(DCFH-DA)探针测定ROS的含量;试剂盒检测SOD和MDA水平;流式细胞术检测细胞凋亡率;Western blot检测Nrf2、HO-1、NQO1蛋白相对表达水平。将裸鼠随机分为对照组、荜茇酰胺(PL)组、圣草酚组、PL+圣草酚组,每组8只;每天腹腔注射PL 2.5 mg/kg或圣草酚40 mg/kg治疗15 d,测定移植瘤体积和重量;DCFH-DA探针测定ROS的含量;试剂盒检测SOD和MDA水平;TUNEL染色检测细胞凋亡;Western blot检测Nrf2、HO-1、NQO1蛋白相对表达水平。结果:圣草酚呈剂量依赖性抑制HL-7702和Huh7细胞活性,圣草酚浓度78μmol/L进行后续细胞实验。与H_(2)O_(2)组相比,H_(2)O_(2)+圣草酚组ROS水平和MDA含量显著上升,SOD活性显著下降,细胞凋亡率显著升高,细胞核Nrf2、HO-1、NOQ1表达显著下调(P<0.05)。与PL组相比,PL+圣草酚组移植瘤体积和重量显著减小,ROS水平和MDA含量显著增加,SOD活性显著降低,细胞凋亡率显著升高,细胞核Nrf2、HO-1、NOQ1表达显著下调(P<0.05)。结论:圣草酚促进氧化应激诱导的肝癌细胞和移植瘤细胞的凋亡,并抑制Nrf2通路激活。Objective:To investigate the mechanism of Eriodictyol on liver cancer cell apoptosis induced by oxidative stress.Methods:The CCK-8 method was used to determine the toxicity of Eriodictyol to cells. Huh7 cells were divided into control group,H_(2)O_(2) group,Eriodictyol group,H_(2)O_(2)+Eriodictyol group,and cells were treated with H_(2)O_(2)0.5 mmol/L or Eriodictyol 78 μmol/L for 48 h. The content of ROS was measured by 2,7-dichlorofluorescein diacetate(DCFH-DA)probe. The SOD and MDA levels were detected by the kit. The rate of apoptosis was detected by flow cytometry. The relative expression levels of Nrf2,HO-1 and NQO1 proteins were detected by Western blot. Nude mice were randomly divided into control group,PL group,Eriodictyol group,and PL+Eriodictyol group. Each nude mouse was injected intraperitoneally with PL 2.5 mg/kg or Eriodictyol 40 mg/kg daily for 15 days to measure the volume and weight of the transplanted tumor. The content of ROS was measured by the DCFH-DA probe,the levels of SOD and MDA were detected by the kit,the apoptosis was detected by TUNEL staining,and the relative expression levels of Nrf2,HO-1 and NQO1 proteins were detected by Western blot.Results:Eriodictyol inhibited the activity of HL-7702 and Huh7 cells in a dose-dependent manner,and the concentration of Eriodictyol was 78 μmol/L for subsequent cell experiments. Compared with the H_(2)O_(2) group,the ROS level and MDA content of the H_(2)O_(2)+Eriodictyol group were significantly increased,the SOD activity was significantly decreased,the apoptosis rate was significantly increased,and the expression of nuclear Nrf2,HO-1,NOQ1 in cells was significantly decreased(P<0.05). Compared with the PL group,the transplanted tumor volume and weight of the PL+Eriodictyol group were significantly reduced,the ROS level and MDA content were significantly increased,the SOD activity was significantly reduced,the apoptosis rate was significantly increased,and the expression of nuclear Nrf2,HO-1 and NOQ1 was significantly decreased(P<0.05).Conclusion:

关 键 词：圣草酚 氧化应激 肝癌 凋亡 NRF2 

分 类 号：R735.7[医药卫生—肿瘤]