二甲双胍抑制铁死亡改善骨关节炎模型大鼠的软骨损伤  

Metformin inhibits ferroptosis and improves cartilage damage in osteoarthritis model rats

作　　者：樊佳欣 贾祥 徐田杰 刘凯楠 郭小玲 张辉[4] 王茜 Fan Jiaxin;Jia Xiang;Xu Tianjie;Liu Kainan;Guo Xiaoling;Zhang Hui;Wang Qian(School of Basic Medical Sciences,North China University of Science and Technology,Tangshan 063210,Hebei Province,China;Key Laboratory of Basic Medicine for Chronic Diseases,Tangshan 063210,Hebei Province,China;College of Basic Medicine,Xingtai Medical College,Xingtai 054000,Hebei Province,China;First Department of Joint Surgery,Tangshan Second Hospital,Tangshan 063000,Hebei Province,China)

机构地区：[1]华北理工大学基础医学院,河北省唐山市063210 [2]河北省慢性疾病基础医学重点实验室,河北省唐山市063210 [3]邢台医学院基础医学部,河北省邢台市054000 [4]唐山市第二医院关节一科,河北省唐山市063000

出　　处：《中国组织工程研究》2025年第30期6398-6408,共11页Chinese Journal of Tissue Engineering Research

基　　金：三三人才工程(C20221117),项目负责人:张辉;河北省2023年度医学科学研究课题(20230215),项目负责人:王茜。

摘　　要：背景:二甲双胍目前被视为治疗2型糖尿病的首选药物。研究发现,二甲双胍可延缓骨关节炎进展,但具体的作用机制仍不明确。目的:评估二甲双胍对骨关节炎大鼠的治疗效果及相关作用机制。方法:①网络药理学:使用CTD、SwissTargetPrediction、GeneCards和OMIM数据库筛选二甲双胍、骨关节炎、铁死亡的潜在共同靶标,导入STRING数据库进行蛋白质-蛋白质相互作用分析得到二甲双胍、骨关节炎和铁死亡的关键靶点。②分子对接:从PDB数据库下载P53及其下游因子SLC7A11蛋白质的PDB格式,将二甲双胍的2D结构式转换为3D结构式,使用Discovery Studio 2019 Client进行二甲双胍和蛋白质的分子对接。③体内实验:取30只雄性SD大鼠,随机分为3组(n=10):空白组不进行任何手术,模型组、二甲双胍组采用改良Hulth法建立骨关节炎模型;术后1 d,二甲双胍组大鼠灌胃二甲双胍200 mg/(kg·d),空白组、模型组灌胃生理盐水,连续给药4周。苏木精-伊红染色、番红O-固绿染色观察大鼠膝关节软骨形态结构并进行Mankin评分;ELISA检测血清中肿瘤坏死因子α、白细胞介素6水平;微板法检测血清中铁死亡相关指标谷胱甘肽、丙二醛、Fe^(2+)水平;免疫荧光染色、Western blot、Real-time qPCR检测大鼠软骨组织中P53、SLC7A11、谷胱甘肽过氧化物酶4、软骨蛋白聚糖、基质金属蛋白酶13的蛋白及mRNA表达。结果与结论:①二甲双胍、骨关节炎与铁死亡交集靶点共96个,经蛋白质-蛋白质相互作用分析发现潜在靶点共有77个,进一步筛选得到核心靶点依次为TP53、AKT1、JUN、白细胞介素6、MYC、白细胞介素1β、肿瘤坏死因子α等。②对接分析结果显示,二甲双胍与P53及其下游因子SLC7A11结合牢固稳定。③模型组大鼠膝关节软骨表面不规则,软骨组织出现缺损,软骨细胞数量减少;相较于模型组,二甲双胍组大鼠膝关节软骨结构损伤明显改善,软骨�BACKGROUND:Metformin is currently considered the first-line medication for the treatment of type 2 diabetes.Metformin may delay the progression of osteoarthritis,but its specific mechanism of action remains unclear.OBJECTIVE:To evaluate the therapeutic effects and the related action mechanisms of metformin on osteoarthritis in rats.METHODS:(1)Network pharmacology:Potential common targets for metformin,osteoarthritis,and ferroptosis were screened using the CTD,SwissTargetPrediction,GeneCards,and OMIM databases.After importing the targets into the STRING database,protein-protein interaction analysis was conducted to identify the key targets for metformin,osteoarthritis,and ferroptosis.(2)Molecular docking:P53 and its downstream factor SLC7A11 protein structures in PDB format were downloaded from the PDB database.The 2D structure of metformin was converted to a 3D structure,and molecular docking of metformin with the proteins was performed using Discovery Studio 2019 Client.(3)In vivo experiments:Thirty male SD rats were randomly divided into three groups(n=10).The blank group did not receive surgery.The osteoarthritis model was established using the modified Hulth method for the model and metformin groups.One day after the surgery,rats in the metformin group were gavaged with metformin 200 mg/kg per day,while the blank and model groups were gavaged with physiological saline.Treatment continued for 4 weeks.Hematoxylin-eosin staining and Safranin O-fast green staining were used to observe the pathological morphology and structure of the knee cartilage,and Mankin scoring was performed.ELISA was used to measure the levels of tumor necrosis factor-αand interleukin-6 in the serum.The microplate method was used to measure serum ferroptosis-related indicators,including glutathione,malondialdehyde,and Fe^(2+).Immunofluorescence staining,western blot assay,and real-time qPCR were used to detect the protein and mRNA expression of P53,SLC7A11,glutathione peroxidase 4,proteoglycans,and matrix metalloproteinase 13 in the cartil

关 键 词：骨关节炎 关节软骨 铁死亡 铁代谢 二甲双胍 P53 SLC7A11 谷胱甘肽过氧化物酶4 工程化组织构建 

分 类 号：R459.9[医药卫生—治疗学] R-332[医药卫生—临床医学] R684.3