海南医科大学学报2025，Vol.31Issue(24)：1851-1861,11.DOI:10.13210/j.cnki.jhmu.20241212.001
电针血清对氧糖剥夺/复氧糖后HT22神经元铁死亡的影响
Effects of electroacupuncture serum on ferroptosis in HT22 neurons after oxygen-glucose deprivation/reoxygenation
摘要
Abstract
Objective:To observe the effect of electroacupuncture serum on ferroptosis in HT22 neurons after oxygen-glucose deprivation/reoxygenation(OGD/R)and to investigate the potential protective mechanisms of electroacupuncture serum on hippo-campal neurons following cerebral ischemia.Methods:A total of 20 male Sprague-Dawley(SD)rats were randomly divided into two groups:A normal group and an electroacupuncture group,with 10 rats in each group.The electroacupuncture group under-went middle cerebral artery occlusion/reperfusion(MCAO/R)modeling using the suture method,followed by 14 days of elec-troacupuncture treatment.After treatment,blood was collected from the abdominal aorta,and electroacupuncture serum was pre-pared at different concentrations.Serum from the normal group was also collected as a control.HT22 hippocampal neurons were used as experimental subjects and were randomly divided into 5 groups:A normal group,a model group,an electroacupuncture se-rum group,an inducer group,and an electroacupuncture serum+inducer group.Except for the normal group,all the other groups underwent 4 hours of OGD,followed by 24 hours of reoxygenation and glucose replenishment,during which they were treated with the corresponding serum.The inducer group and the electroacupuncture serum+inducer group received 10 μmol/L of the ferroptosis inducer Erastin 24 hours before OGD.Cell viability and cytotoxicity were assessed using the CCK-8 and LDH assays,while apoptosis was evaluated via flow cytometry.Mitochondrial structure was observed under a transmission electron micro-scope,reactive oxygen species(ROS)levels were measured using a fluorescence probe,and intracellular Fe²⁺,glutathione(GSH),and malondialdehyde(MDA)levels were detected using colorimetric kits.Western blot analysis was employed to detect the relative expression of transferrin receptor 1(TFR1),ferritin heavy chain polypeptide 1(FTH1),nuclear receptor coactivator 4(NCOA4),solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),acyl-CoA synthetase long-chain family member 4(ACSL4),and lipoxygenase(LOX).Results:Electroacupuncture serum at concentrations of 5%,10%,15%,and 20%improved cell morphology and reduced cytotoxicity(P<0.01),with 10%yielding the best results.Compared to the normal group,the model group exhibited decreased cell viability(P<0.01),increased cytotoxicity and apoptosis(P<0.01).Transmission electron microscope showed remarkable mitochondrial swelling,cristae disruption or disappearance,elevated Fe2+,MDA,and ROS levels(P<0.01),reduced GSH level(P<0.01),decreased expressions of FTH1,SLC7A11,and GPX4(P<0.05),and increased expressions of TFR1,NCOA4,ACSL4,and LOX(P<0.05).Compared to the model group,the elec-troacupuncture serum group showed increased cell viability(P<0.01),reduced cytotoxicity and apoptosis(P<0.05),relatively intact mitochondrial structure,decreased Fe2+,MDA,and ROS levels(P<0.01),increased GSH level(P<0.05),elevated ex-pressions of FTH1,SLC7A11,and GPX4(P<0.05),and reduced expressions of TFR1,NCOA4,ACSL4,and LOX(P<0.05).The inducer group,compared to the model group,exhibited decreased cell viability(P<0.01),increased cytotoxicity and apoptosis(P<0.01),more pronounced mitochondrial swelling and structural damage,elevated Fe2+,MDA,and ROS level(P<0.01),reduced GSH levels(P<0.01),decreased FTH1,SLC7A11,and GPX4 expressions(P<0.05),and increased TFR1,NCOA4,ACSL4,and LOX expressions(P<0.05).Compared to the inducer group,the electroacupuncture serum+inducer group showed increased cell viability(P<0.01),reduced cytotoxicity and apoptosis(P<0.01),improved mitochondrial morphol-ogy,decreased Fe2+,MDA,and ROS levels(P<0.01),increased GSH level(P<0.01),elevated FTH1,SLC7A11,and GPX4 expressions(P<0.05),and reduced TFR1,NCOA4,ACSL4,and LOX expressions(P<0.05).Conclusion:Elec-troacupuncture serum effectively inhibits ferroptosis in HT22 neurons following OGD/R,thereby playing a significant role in miti-gating hippocampal neuronal damage after cerebral ischemia.关键词
电针血清/氧糖剥夺/复氧糖(OGD)/R/HT22神经元/铁死亡Key words
Electroacupuncture serum/OGD/R/HT22 neurons/Ferroptosis分类
医药卫生引用本文复制引用
CHEN Lulu,WANG Huiling,LI Ruiqing,FENG Xiaodong,RUAN Xiaodi,GAO Jing,CHEN Bofei,SU Kaiqi,Lyu Zhuan,FANG Lu,AN Yuqi,WU Mingli..电针血清对氧糖剥夺/复氧糖后HT22神经元铁死亡的影响[J].海南医科大学学报,2025,31(24):1851-1861,11.基金项目
国家自然科学基金(82174473) （82174473）
国家自然科学基金联合基金(U2004131) （U2004131）
河南省科技攻关计划项目(232102311210) （232102311210）
河南省科技研发计划联合基金(232301420072) This study was supported by the National Natural Science Foundation of China(82174473) （232301420072）
National Natural Science Foundation of China Joint Fund(U2004131) （U2004131）
Science and Technology Research and Development Program of Henan Province(232102311210) （232102311210）
Joint Fund for Science and Technology Research and Development Plan of Henan Province(232301420072) （232301420072）
