淫羊藿苷对激素诱导大鼠骨微血管内皮细胞铁死亡的影响  

Effect of Icariin on Steroid-induced Ferroptosis in Rat Bone Microvascular Endothelial Cells

作　　者：汤建成 张悦 姜睿琛 岳峥嵘 李铭 张亚奇 尹泽陶 王卫国[4] TANG Jiancheng;ZHANG Yue;JIANG Ruichen;YUE Zhengrong;LI Ming;ZHANG Yaqi;YIN Zetao;WANG Weiguo(Graduate School,Beijing University of Chinese Medicine,Beijing 100029,China;The First Affiliated Hospital,Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014010,China;Peking University Health Science Center,Beijing 100191,China;China-Japan Friendship Hospital,Beijing 100029,China)

机构地区：[1]北京中医药大学研究生院,北京100029 [2]内蒙古科技大学包头医学院第一附属医院,内蒙古包头014010 [3]北京大学医学部,北京100191 [4]中日友好医院,北京100029

出　　处：《中国实验方剂学杂志》2025年第5期131-140,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81972107)。

摘　　要：目的:探究淫羊藿苷(ICA)对激素诱导骨微血管内皮细胞(BMECs)铁死亡的影响。方法:选取大鼠骨BMECs,用500 mg·L^(-1)氢化可的松干预1.5 h建立BMECs铁死亡模型,分为空白组、激素组(500 mg·L^(-1)氢化可的松)、淫羊藿苷组(500 mg·L^(-1)氢化可的松+34 mg·L^(-1)淫羊藿苷)、铁死亡激动剂组(500 mg·L^(-1)氢化可的松+34 mg·L^(-1)淫羊藿苷+2.7 mg·L^(-1)erastin)。用细胞增殖与活性检测(CCK-8)法检测细胞活力;用相关试剂盒检测细胞亚铁离子、谷胱甘肽(GSH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、活性氧(ROS)水平;蛋白免疫印迹法(Western blot)检测铁死亡相关蛋白谷胱甘肽过氧化物酶4(GPX4)和铁蛋白轻链(FTL)、转铁蛋白受体(sTfR),自噬相关蛋白微管相关蛋白轻链3B(LC3B)、自噬效应蛋白1(Beclin1),凋亡相关蛋白B细胞淋巴瘤-2(Bcl-2)、胱天蛋白酶-3(Caspase-3)表达水平。结果:500 mg·L^(-1)氢化可的松干预1.5 h可有效诱导BMECs铁死亡的发生,铁死亡水平可随着干预时间的增加达到峰值。在细胞抗氧化能力方面,与空白组比较,激素组细胞活力、GSH显著降低,ROS、SOD、MDA、亚铁离子水平显著增多(P<0.01);与激素组比较,淫羊藿苷组细胞活力、GSH显著增多,ROS、SOD、MDA、亚铁离子水平显著降低(P<0.01);与淫羊藿苷组比较,铁死亡激动剂组细胞活力、GSH显著降低,ROS、SOD、MDA、亚铁离子水平显著增高(P<0.01)。在细胞铁死亡和自噬关系方面,与空白组比较,激素组Beclin1、LC3B、sTfR、FTL表达水平升高,GPX4表达水平降低(P<0.01);与激素组比较,淫羊藿苷组细胞内LC3B、sTfR、FTL表达水平降低,Beclin1、GPX4表达水平显著升高(P<0.01);与淫羊藿苷组比较,雷帕霉素组LC3B、sTfR、FTL表达水平升高,Beclin1、GPX4表达水平显著降低(P<0.01)。在细胞铁死亡与凋亡方面,与空白组比较,激素组FTL、sTfR、Caspase-3表达水平升高,GPX4、Bcl-2表达水平显著降低(P<0.01);与激素�Objective:To investigate the effect of icariin(ICA)on steroid-induced ferroptosis in bone microvascular endothelial cells(BMECs).Methods:Rat BMECs were selected and treated with 500 mg·L^(-1) hydrocortisone for 1.5 h to establish a ferroptosis model of BMECs.The experimental cells were divided into a blank group,hormone group(500 mg·L^(-1) hydrocortisone),ICA group(500 mg·L^(-1) hydrocortisone+34 mg·L^(-1) ICA),and ferroptosis agonist group(500 mg·L^(-1) hydrocortisone+34 mg·L^(-1) ICA+2.7 mg·L^(-1) erastin).Cell viability was detected by CCK-8.The levels of ferrous ion,glutathione(GSH),malondialdehyde(MDA),superoxide dismutase(SOD),and reactive oxygen species(ROS)were detected by related kit species.The ferroptosisrelated proteins,such as glutathione peroxidase 4(GPX4),ferritin light chain(FTL),and transferrin receptor protein1(sTfR)were detected by Western blot,as well as autophagy-related proteins including microtubule-associated protein 1 light chain 3B(LC3B),Beclin1,B-cell lymphoma-2(Bcl-2),and Caspase-3.Results:500 mg·L^(-1) hydrocortisone intervention for 1.5 h could effectively induce ferroptosis in BMECs,and ferroptosis levels could reach a peak as the intervention continued.In terms of cellular antioxidant capacity,compared with those in the blank group,the cell vitality,GSH in the hormone group decreased significantly,and the levels of ROS,SOD,MDA,and ferrous ions were significantly increased(P<0.01).Compared with those in the hormone group,the cell viability,GSH were significantly increased,and the levels of ROS,SOD,MDA,and ferrous ions were decreased in the ICA group(P<0.01).Compared with those in the ICA group,the cell vitality,GSH in the ferroptosis agonist group decreased significantly,and the levels of ROS,SOD,MDA,and ferrous ions increased significantly(P<0.01).In terms of the relationship between ferroptosis and autophagy,compared with the blank group,the hormone group had significantly increased expression levels of LC3B,sTfR,Beclin1,and FTL and significantly decreased expression level

关 键 词：淫羊藿苷 激素性股骨头坏死 骨微血管内皮细胞 自噬 铁死亡 

分 类 号：R2-031[医药卫生—中西医结合] R681[医药卫生—中医学] R285.5