Esculetin,a natural dihydroxy coumarin derived from the Chinese herbal medicine Cortex Fraxini,has demonstrated significant pharmacological activities,including anticancer properties.Ferroptosis,an iron-dependent form...Esculetin,a natural dihydroxy coumarin derived from the Chinese herbal medicine Cortex Fraxini,has demonstrated significant pharmacological activities,including anticancer properties.Ferroptosis,an iron-dependent form of regulated cell death,has garnered considerable attention due to its lethal effect on tumor cells.However,the exact role of ferroptosis in esculetin-mediated anti-hepatocellular carcinoma(HCC)effects remains poorly understood.This study investigated the impact of esculetin on HCC cells both in vitro and in vivo.The findings indicate that esculetin effectively inhibited the growth of HCC cells.Importantly,esculetin promoted the accumulation of intracellular Fe^(2+),leading to an increase in ROS production through the Fenton reaction.This event subsequently induced lipid peroxidation(LPO)and triggered ferroptosis within the HCC cells.The occurrence of ferroptosis was confirmed by the elevation of malondialdehyde(MDA)levels,the depletion of glutathione peroxidase(GSH-Px)activity,and the disruption of mitochondrial morphology.Notably,the inhibitor of ferroptosis,ferrostatin-1(Fer-1),attenuated the anti-tumor effect of esculetin in HCC cells.Furthermore,the findings revealed that esculetin inhibited the Nrf2-xCT/GPx4 axis signaling in HCC cells.Overexpression of Nrf2 upregulated the expression of downstream SLC7A11 and GPX4,consequently alleviating esculetin-induced ferroptosis.In conclusion,this study suggests that esculetin exerts an anti-HCC effect by inhibiting the activity of the Nrf2-xCT/GPx4 axis,thereby triggering ferroptosis in HCC cells.These findings may contribute to the potential clinical use of esculetin as a candidate for HCC treatment.展开更多
基金supported by the Natural Science Foundations of Fujian Province(Nos.2021J05063 and 2023J01541)a startup grant for High-level Talents of Fujian Medical University(No.XRCZX2021014)。
文摘Esculetin,a natural dihydroxy coumarin derived from the Chinese herbal medicine Cortex Fraxini,has demonstrated significant pharmacological activities,including anticancer properties.Ferroptosis,an iron-dependent form of regulated cell death,has garnered considerable attention due to its lethal effect on tumor cells.However,the exact role of ferroptosis in esculetin-mediated anti-hepatocellular carcinoma(HCC)effects remains poorly understood.This study investigated the impact of esculetin on HCC cells both in vitro and in vivo.The findings indicate that esculetin effectively inhibited the growth of HCC cells.Importantly,esculetin promoted the accumulation of intracellular Fe^(2+),leading to an increase in ROS production through the Fenton reaction.This event subsequently induced lipid peroxidation(LPO)and triggered ferroptosis within the HCC cells.The occurrence of ferroptosis was confirmed by the elevation of malondialdehyde(MDA)levels,the depletion of glutathione peroxidase(GSH-Px)activity,and the disruption of mitochondrial morphology.Notably,the inhibitor of ferroptosis,ferrostatin-1(Fer-1),attenuated the anti-tumor effect of esculetin in HCC cells.Furthermore,the findings revealed that esculetin inhibited the Nrf2-xCT/GPx4 axis signaling in HCC cells.Overexpression of Nrf2 upregulated the expression of downstream SLC7A11 and GPX4,consequently alleviating esculetin-induced ferroptosis.In conclusion,this study suggests that esculetin exerts an anti-HCC effect by inhibiting the activity of the Nrf2-xCT/GPx4 axis,thereby triggering ferroptosis in HCC cells.These findings may contribute to the potential clinical use of esculetin as a candidate for HCC treatment.
文摘目的探究枸杞多糖(Lycium barbarum polysaccharides,LBP)调控核因子E2相关因子2(nuclear factor E2 related factor 2,Nrf2)/谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)通路对缺氧/复氧(hypoxia/reoxygenation,H/R)诱导的心肌细胞铁死亡的影响。方法设置对照组、模型组及LBP低、中、高剂量(25、50、100 mg/L)组和LBP(100mg/L)+Nrf2抑制剂ML385(5μmol/L)组。构建H9c2细胞H/R模型,并给予相应药物处理。采用CCK-8检测细胞活力;采用TUNEL染色检测细胞凋亡;采用试剂盒检测细胞Fe^(2+)、谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)水平及乳酸脱氢酶(lactate dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)活性;采用二氢乙锭(dihydroethidium,DHE)荧光探针检测细胞活性氧(reactive oxygen species,ROS)水平;采用透射电镜观察细胞线粒体形态;采用Western blotting检测转铁蛋白受体1(transferrin receptor 1,Tf R1)、二价金属离子转运蛋白1(divalent metal transporter 1,DMT1)、铁蛋白重链1(ferritin heavy chain 1,FTH1)、酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long-chain family member 4,ACSL4)、Kelch样ECH相关蛋白1(Kelch like ECH associated protein1,Keap1)、Nrf2、血红素氧合酶-1(heme oxygenase-1,HO-1)、溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)、GPX4蛋白表达。结果与对照组比较,模型组细胞活力显著降低(P<0.001),Fe^(2+)、MDA、ROS水平和LDH活性显著升高(P<0.001),GSH水平和SOD、CAT活性显著降低(P<0.001),线粒体缩短、膜密度增高且嵴数量减少,TfR1、Nrf2、HO-1、SLC7A11、GPX4蛋白表达水平显著降低(P<0.001),DMT1、FTH1、ACSL4、Keap1蛋白表达水平显著升高(P<0.001)。与模型组比较,LBP组细胞活力显著升高(P<0.01),Fe^(2+)、MDA、ROS水平和LDH活性显著降低(P<0.05、0.01、0.001),GSH水平和SOD、CAT活性显著升高(P<0.05、0.01、0.001),线粒体形态较为规则,TfR1、Nrf2、HO-1、SLC7A11、GPX4蛋白表达水平显著升高(P<0.01、0.001),DMT1、FTH1、ACSL4、Keap1蛋白表达水平显著降低(P<0.05、0.01、0.001);而ML385能够抑制枸杞多糖对细胞铁死亡的改善作用(P<0.05、0.01、0.001)。结论枸杞多糖可能通过激活Nrf2/GPX4通路改善H/R诱导的心肌细胞铁死亡。
