青蒿琥酯抑制Nrf2/HIF-1α促进缺氧肝癌细胞增敏作用  

Artesunate Enhances the Sensitization of Hepatocellular Carcinoma Cells by Inhibiting Nrf2/HIF-1αin Hypoxia

作　　者：陈源 王美慧 李伟[3] 许学芬[1] CHEN Yuan;WANG Meihui;LI Wei;XU Xuefen(School of Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Integrated Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学医学院,江苏南京210023 [2]南京中医药大学药学院,江苏南京210023 [3]南京中医药大学中西医结合学院,江苏南京210023

出　　处：《南京中医药大学学报》2025年第2期243-250,共8页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(81703542);南京中医药大学自然科学基金(NZY81703542)。

摘　　要：目的探讨缺氧微环境下青蒿琥酯增加肝癌细胞对索拉非尼的敏感性及其相关机制。方法建立肝癌细胞缺氧耐药模型,采用MTT检测缺氧微环境下青蒿琥酯对肝癌细胞抑制率的影响及主要死亡方式。试剂盒检测胞内GSH、ROS、脂质过氧化物MDA水平、铁离子水平;JC-1检测线粒体膜电位变化;Western blot和qPCR检测加入青蒿琥酯后铁死亡相关基因和蛋白的表达。结果缺氧微环境下肝癌细胞对索拉非尼敏感性下降,青蒿琥酯能有效增强这一敏感性;青蒿琥酯通过增加肝癌细胞ROS水平(P<0.05,P<0.01,P<0.001)促进索拉非尼诱导的铁死亡发生。进一步分析揭示青蒿琥酯能抑制Nrf2的mRNA和蛋白水平(P<0.05,P<0.01,P<0.001)并下调HIF-1α的表达(P<0.05,P<0.01,P<0.001)。结论缺氧微环境下青蒿琥酯可能通过抑制Nrf2调控HIF-1α蛋白水平增加胞内氧化应激水平,诱导肝癌细胞铁死亡的发生,从而增加肝癌细胞对索拉非尼敏感性。OBJECTIVE To investigate the mechanism of artesunate increasing the sensitivity of hepatocellular carcinoma(HCC)cells to sorafenib in hypoxic microenvironment.METHODS Firstly,hypoxia-resistant HCC cell models were established.NAC(a ROS scavenger),necrostatin-1(a necrosis inhibitor),and ferrostatin-1(a ferroptosis inhibitor)were added to examine the effects of artesunate on the inhibition rate of HCC cells and the primary modes of cell death in hypoxic microenvironment by MTT assays.Intracellular levels of GSH,ROS,lipid peroxidation product MDA,and iron ions were measured using corresponding kits.Mitochondrial membrane potential changes were assessed using JC-1 staining.Western blot and qPCR were performed to detect the expression of ferroptosis-related proteins after the addition of artesunate.RESULTS The sensitivity of HCC cells to sorafenib decreased under hypoxic microenvironment,but artesunate significantly enhanced this sensitivity.Further analysis revealed that artesunate promoted sorafenib-induced ferroptosis by increasing ROS levels in HCC cells(P<0.05,P<0.01,P<0.001).Additionally,artesunate was found to inhibit Nrf2 mRNA and protein levels(P<0.05,P<0.01,P<0.001)and downregulate HIF-1αexpression(P<0.05,P<0.01,P<0.001).CONCLUSION Artesunate increases intracellular oxidative stress by inhibiting Nrf2 and HIF-1αprotein levels,subsequently induces ferroptosis and enhances the sensitivity of HCC cells to sorafenib in hypoxic microenvironment.

关 键 词：青蒿琥酯 铁死亡 肝癌细胞 索拉非尼 ROS NRF2 HIF-1Α 

分 类 号：R285.5[医药卫生—中药学]