基于ROS/GSH/GPX4轴探讨骨痹饮对膝骨关节炎模型小鼠软骨组织铁死亡的影响  

Effects of Gubi Decoction on Ferroptosis in Cartilage Tissues of Mice with Knee Osteoarthritis Based on ROS/GSH/GPX4 Axis

作　　者：崔龙慷 余阳 鲍正生 沈高波 张兵兵 吴连国 CUI Long-kang;YU Yang;BAO Zheng-sheng;SHEN Gao-bo;ZHANG Bing-bing;WU Lian-guo(Orthopedics and Traumatology Department,The Second Affiliated Hospital of Zhejiang Chinese Medicine University,Hangzhou 310005;The Second Clinical School,Zhejiang Chinese Medicine University,Hangzhou 310053)

机构地区：[1]浙江中医药大学附属第二医院骨伤科,杭州310005 [2]浙江中医药大学第二临床医学院,杭州310053

出　　处：《中国中西医结合杂志》2025年第3期313-320,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：浙江省“尖兵”“领雁”研发攻关计划(No.2024C03213);浙江省中医药科技计划项目(No.2024ZL514);浙江中医药大学科研项目资助(No.2023JKZKTS41)。

摘　　要：目的基于活性氧(ROS)/谷胱甘肽(GSH)/谷胱甘肽过氧化物酶4(GPX4)轴探究中药汤剂骨痹饮抑制膝骨关节炎(KOA)软骨组织铁死亡的作用机制。方法将50只7周龄C57BL/6雄性小鼠随机分为假手术组、模型组、阳性药组及骨痹饮低、高剂量组,每组10只。除假手术组外,其余各组小鼠予右旋糖酐铁120 mg/kg腹腔注射,每周1次,持续8周;注射右旋糖酐铁2周后,行左膝内侧半月板失稳(DMM)手术。阳性药组予塞来昔布溶液100 mg/kg灌胃,每天1次,持续8周;骨痹饮低、高剂量组分别给予骨痹饮8.65、17.29 g/kg灌胃,每天1次,持续8周。检测小鼠左膝关节直径和机械痛阈值;HE染色和番红O-固绿染色观察关节软骨组织病理变化,并进行国际骨关节炎研究学会(OARSI)评分;普鲁士蓝染色评估膝关节的铁沉积情况;生化试剂盒检测软骨组织中GSH、ROS的表达水平;Western Blot检测软骨组织中GPX4、铁蛋白重链1(FTH1)蛋白的表达水平。结果骨痹饮和塞来昔布均能显著降低小鼠左膝关节直径(P<0.05),提高机械痛阈值(P<0.05),明显改善软骨组织病理形态损伤,减少铁沉积。与模型组比较,骨痹饮低、高剂量组软骨组织GSH含量、FTH1、GPX4蛋白表达水平增加,ROS含量显著降低(P<0.05);与骨痹饮低剂量组比较,骨痹饮高剂量组软骨组织GSH含量、FTH1、GPX4蛋白表达增加,ROS含量显著降低(P<0.05)。结论骨痹饮可有效缓解KOA模型小鼠膝关节肿胀及疼痛,改善软骨组织损伤,其作用机制可能与调控ROS/GSH/GPX4通路抑制软骨细胞铁死亡有关。Objective To observe the possible mechanism of Gubi Decoction on ferroptosis in cartilage tissues of mice with knee osteoarthritis(KOA)based on the reactive oxygen species(ROS)/glutathione(GSH)/glutathione peroxidase 4(GPX4)axis.Methods A total of 50 seven-week male C57BL/6 mice were randomly divided into the sham-operated,model,positive drug,and low-,high-dose Gubi Decoction groups,10 in each group.Except for the sham-operated group,mice were administered iron dextran intraperitoneally(120 mg/kg)once a week for eight weeks.After the injection of iron dextran for 2 weeks,KOA was induced by destabilizing the medial meniscus(DMM)in the left knee joints.Mice in the positive drug group were administered with celecoxib intragastrically(100 mg/kg)once a day for eight weeks.Mice in the low-and high-dose of Gubi Decoction groups were administered with Gubi Decoction(8.65,17.29 g/kg)intragastrically once a day for eight weeks.After all the interventions,left knee joint diameter and mechanical pain threshold were measured.The pathological changes of cartilage tissue was observed by HE staining and Safranin-O/Fast Green staining,and the International Society for Osteoarthritis Research(OARSI)score was performed.Prussian blue staining was used to evaluate the iron deposition in the knee joint.Biochemical kit was used to detect the expression levels of GSH and ROS in cartilage tissues.Western blot was used to detect the expression levels of GPX4 and ferritin heavy chain 1(FTH1)proteins in cartilage tissues.Results Gubi Decoction and celecoxib significantly decreased the left knee joint diameter,the OARSI score and the iron deposition positive area of mice(P<0.05);increased the paw withdrawal threshold(P<0.05);improved the cartilage tissue damage.Compared with model group,GSH content,FTH1 and GPX4 protein expression levels in cartilage tissue of low and high dose Gubi Decoction groups increased,and ROS content decreased(P<0.05).Compared with low-dose Gubi Decoction group,GSH content,FTH1 and GPX4 protein expressions increased

关 键 词：膝骨关节炎 骨痹饮 软骨组织 铁死亡 活性氧/谷胱甘肽/谷胱甘肽过氧化物酶4轴 中药复方 中西医结合 

分 类 号：R285.5[医药卫生—中药学]