摘要
目的探讨肉桂酸改善缺氧/复氧(A/R)诱导心肌细胞损伤的作用机制。方法将H9c2细胞分组为对照组、A/R组、肉桂酸低和高剂量组、HIF-1α激活剂(DMOG)组、肉桂酸高剂量+HIF-1α抑制剂(IDF-11774)组。CCK-8法检测细胞活力;ELISA检测H9c2细胞上清中乳酸脱氢酶(LDH)、肌酸激酶同工酶(CK-MB)活性;透射电镜观察H9c2细胞线粒体超微结构;2',7'-二氯二氢荧光素二乙酸酯(DCFH-DA)荧光探针检测H9c2细胞中活性氧(ROS)荧光强度;试剂盒检测H9c2细胞中丙二醛(MDA)、超氧化物歧化酶(SOD)活性;FerroOrange荧光探针检测H9c2细胞中亚铁(Fe^(2+))荧光强度;Western blot检测H9c2细胞中HIF-1α、SLC7A11、GPX4蛋白。结果与对照组比较,A/R组H9c2细胞中线粒体结构萎缩,嵴破坏严重,部分外膜破裂,H9c2细胞活力、SOD活性及HIF-1α、SLC7A11、GPX4蛋白降低,细胞上清中LDH、CK-MB活性、细胞中ROS荧光强度、MDA水平及Fe^(2+)荧光强度升高(P<0.05);与A/R组比较,肉桂酸低和高剂量组、DMOG组H9c2细胞线粒体结构破坏程度减轻,H9c2细胞活力、SOD活性及HIF-1α、SLC7A11、GPX4蛋白升高,细胞上清中LDH、CK-MB活性、细胞中ROS荧光强度、MDA水平及Fe^(2+)荧光强度降低(P<0.05);IDF-11774减弱了高剂量肉桂酸对A/R诱导的H9c2细胞铁死亡的抑制作用以及对细胞损伤的改善作用。结论肉桂酸可能通过激活HIF-1α/SLC7A11/GPX4通路抑制A/R诱导的H9c2细胞铁死亡,减轻细胞损伤。
Objective To explore the mechanism of cinnamic acid(CA)in improving anoxia/reoxygenation(A/R)-induced myocardial cell injury.Methods H9c2 cells were assigned into control group,A/R group,low-dose and high-dose groups of cinnamic acid,HIF-1αactivator(DMOG)group,and high-dose cinnamic acid+HIF-1αinhibitor(IDF-11774)group.Cell viability was assessed using the CCK-8 assay.Levels of lactate dehydrogenase(LDH)and creatine kinase-MB(CK-MB)in the supernatant were measured by ELISA.Mitochondrial ultrastructure was observed via transmission electron microscopy.Intracellular reactive oxygen species(ROS)and ferrous iron(Fe^(2+))levels were detected using 2′,7′-dichlorodihydrofluorescein diacetate(DCFH-DA)and FerroOrange fluorescent probes,respectively.Malondialdehyde(MDA)levels and superoxide dismutase(SOD)activity were determined using commercial kits.Protein expression of HIF-1α,SLC7A11,and GPX4 was analyzed by Western blot.Results Compared with the control group,the A/R group exhibited mitochondrial atrophy,severe cristae disruption,and partial outer membrane rupture.Additionally,cell viability and SOD activity,along with HIF-1α,SLC7A11,and GPX4 protein levels,were significantly decreased,whereas LDH and CK-MB release,ROS production,MDA levels,and Fe^(2+)accumulation were significantly increased(P<0.05).Compared with the A/R group,the low and high-dose cinnamic acid groups,and DMOG group showed reduced mitochondrial structural injury in H9c2 cells,raised cell viability,SOD activity,HIF-1α,SLC7A11,and GPX4 proteins,and decreased cell supernatant LDH,CK-MB levels,cell ROS fluorescence intensity,MDA and Fe^(2+)fluorescence intensity(P<0.05).IDF-11774 attenuated the inhibitory effect of high-dose cinnamic acid on A/R induced ferroptosis in H9c2 cells and its improvement effect on cell injury.Conclusions Cinnamic acid alleviates A/R-induced cardiomyocyte injury by inhibiting ferroptosis,likely through activation of the HIF-1α/SLC7A11/GPX4 signaling pathway.
作者
韩丽婷
王琰
陈星
马佳
刘洋
HAN Liting;WANG Yan;CHEN Xing;MA Jia;LIU Yang(Department of Cardiac Rehabilitation,Zhengzhou Seventh People′s Hospital,Zhengzhou 450008,China;Department of Cardiology,Zhengzhou Seventh People′s Hospital,Zhengzhou 450008,China)
出处
《基础医学与临床》
2026年第4期540-546,共7页
Basic and Clinical Medicine
基金
郑州市医疗卫生领域科技创新指导计划(2024YLZDJH222)。
关键词
肉桂酸
缺氧/复氧
心肌细胞
铁死亡
cinnamic acid
anoxia/reoxygenation
myocardial cells
ferroptosis
