木犀草素抑制NF-κB增敏TRAIL诱导肝癌HepG2细胞凋亡作用的研究  

Effect of Luteolin Sensitized Human Live Cancer HepG2 Cells to TRAIL-induced Apoptosis by Inhibiting the Activity of NF-κB

作　　者：吴文明 侯雄军[1] 何洁 胡建新[1] WU Wenming;HOU Xiongjun;HE Jie;HU Jianxin(Department of Pharmacy,Jiangxi Provincial People's Hospital(The First Affiliated Hospital of Nanchang Medical College),Nanchang 330006,China)

机构地区：[1]江西省人民医院(南昌医学院第一附属医院)药学部,江西百南昌330006

出　　处：《现代中药研究与实践》2023年第3期16-20,共5页Research and Practice on Chinese Medicines

基　　金：国家自然科学基金(82004063);江西省自然科学基金(20192BAB215056);江西省卫健委普通科技计划项目(20181009);江西省中医药管理局中药制剂临床安全监测和转化应用重点研究室项目;江西省中医药中青年骨干人才(第二批)培养计划。

摘　　要：目的 研究木犀草素抑制NF-κB增敏肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导肝癌细胞HepG2凋亡的作用。方法 体外培养人肝癌HepG2细胞,实验分为空白对照组、木犀草素组、TRAIL组、木犀草素+TRAIL组;采用CCK8法分析细胞存活率;流式细胞术检测细胞凋亡情况和细胞周期;Western Blotting法检测IκBα、p-IκBα和NF-κB p65蛋白表达情况。结果 木犀草素(50μmol/L)、TRAIL(20 ng/mL)以及木犀草素+TRAIL作用于HepG2细胞24 h,细胞的凋亡率分别为(11.67±0.20)%、(6.61±0.15)%和(32.93±0.63)%;流式细胞凋亡和周期检测结果显示木犀草素显著增加细胞凋亡,其中,G_(0)/G_(1)期细胞减少,S期细胞增加;TRAIL组细胞变化无统计学意义;木犀草素+TRAIL组细胞凋亡更甚,其中,G_(0)/G_(1)和G_(2)/M期细胞减少,S期增加,细胞停滞于S期;Western Blotting检测结果显示IκBα、p-IκBα和NF-κB p65蛋白在木犀草素组细胞中表达下降,且在木犀草素+TRAIL组中下降更为明显。结论 20 ng/mL TRAIL对HepG2细胞凋亡无显著影响,50μmol/L亚细胞毒性的木犀草素能够增敏TRAIL对HepG2细胞的凋亡作用,主要作用在S期,其作用机制可能与抑制NF-κB的表达有关。Objective To study the effect of luteolin sensitized human live cancer HepG2 cells to TRAIL-induced apoptosis by inhibiting the activity of NF-κB.Methods Hep G2 cells were cultured in vitro and divided into four groups:control group,luteolin group,TRAIL group and luteolin + TRAIL group.Cell survival rate was analyzed by CCK8 method.Cell apoptosis and cell cycle were detected by flow cytometry.The expressions of IκBα,p-IκBα and NF-κB p65 were detected by Western Blotting method.Results The apoptosis rates of HepG2 cells treated with luteolin(50 μmol/L),TRAIL(20 ng/mL) and luteolin + TRAIL for 24 h were(11.67 ± 0.20)%,(6.61 ± 0.15)% and(32.93 ± 0.63)%,respectively.Flow cytometry showed that luteolin significantly increased cell apoptosis,and the number of cells in G0/G1 phase decreased,while the number of cells in S phase increased.There was no significant difference in cell changes in TRAIL group.In the luteolin + TRAIL group,the cell apoptosis was more severe,and the number of cells in G0/G1and G2/M phase was decreased,the number of cells in S phase was increased,and the cells were stagnated in S phase.The results of Western Blotting detection showed that the expressions of IκBα,p-IκBα and NF-κB p65 protein decreased in luteolin group,and the decrease was more significant in luteolin + TRAIL group.Conclusion 20 ng/mL TRAIL has no significant effect on apoptosis of HepG2 cells.Luteolin with subcytotoxicity of 50 μmol/L sensitivities TRAIL on apoptosis of HepG2 cells,mainly in S phase.The mechanism may be related to the inhibition of NF-κB expression.

关 键 词：HepG2细胞 木犀草素 TRAIL NF-ΚB 凋亡机制 

分 类 号：R285.5[医药卫生—中药学]