益肾逐瘀通络汤对膝骨关节炎大鼠软骨细胞焦亡和铁死亡的影响  

Effect of Yishen Zhuyu Tongluo Decoction on chondrocyte pyroptosis and ferroptosis in rats with knee osteoarthritis

作　　者：王静雅 闫康 李继安 王萌 杨雨旸 虞跃跃 WANG Jingya;YAN Kang;LI Jian;WANG Meng;YANG Yuyang;YU Yueyue(College of Traditional Chinese Medicine,North China University of Science and Technology,Tangshan 063210,China;Key Laboratory of Integrated Traditional Chinese and WesternMedicine,Hebei Province,Tangshan 063210;College of Traditional Chinese Medicine,Hebei University,Baoding 071000)

机构地区：[1]华北理工大学中医学院,河北唐山063210 [2]河北省中西医结合重点实验室,河北唐山063210 [3]河北大学中医学院,河北保定071000

出　　处：《中国比较医学杂志》2025年第1期49-58,共10页Chinese Journal of Comparative Medicine

基　　金：河北省国际科技合作基地建设专项(19392507D);国家科技部对发展中国家科技援助项目(KY201904005)。

摘　　要：目的探究益肾逐瘀通络汤对膝骨关节炎(knee osteoarthritis,KOA)大鼠软骨细胞焦亡和铁死亡的影响。方法40只SD大鼠随机分为假手术组(10只)和造模组(30只);造模组大鼠右后肢膝关节均采用前交叉韧带离断术(anterior cruciate ligament-transection,ACLT)制作KOA模型,抽屉试验检测是否到位,造模操作后,驱赶大鼠每天强制活动30 min以诱导KOA模型,造模4周后大鼠出现跛行、跳跃动作,且苏木素-伊红(HE)染色出现软骨表层缺损变形和炎细胞浸润确认造模成功,造模成功后,大鼠随机分为模型组(n=8)、塞来昔布组(n=8)和益肾逐瘀通络汤组(n=8)。各组大鼠给予生理盐水或药物灌胃干预,其中,假手术组和模型组大鼠灌胃10 mL/kg的生理盐水,塞来昔布组大鼠灌胃12 mg/kg的塞来昔布水溶液,益肾逐瘀通络汤组大鼠灌胃5.4 g/kg的中药水煎液,每天1次,连续8周。干预完成后,麻醉大鼠,腹主动脉采血分离血清,分离膝关节,每组取3份膝关节固定于多聚甲醛,其他分离关节软骨组织备用。使用HE、番红固绿染色观察大鼠关节面软骨病理改变;micro-CT分析骨微结构;ELISA检测血清白细胞介素(interleukin,IL)-1β、IL-18的蛋白表达量;逆转录定量PCR(reverse transcription quantitative real-time PCR,RT-qPCR)检测细胞焦亡和铁死亡相关基因NLRP3、ASC、Caspase-1、GSDMD、IL-1β、IL-18、ACSL-4、FTH-1、GPX-4和COX-2的mRNA相对表达水平。免疫组化检测NLRP3、Caspase-1、COX-2蛋白表达情况。结果与假手术组比较,模型组大鼠软骨组织表层缺损变形,整体各层细胞排列紊乱,番红固绿染色丢失较多,骨小梁稀疏、分布紊乱;血清炎症因子IL-1β和IL-18升高(P<0.01);软骨组织NLRP3、ASC、Caspase-1、GSDMD、IL-1β、IL-18、ACSL-4和COX-2 mRNA表达量均升高(均P<0.01),FTH-1和GPX-4 mRNA表达量均下降(P<0.01)。与模型组比较,塞来昔布和益肾逐瘀通络汤组大鼠软骨表面较完整且平滑,细胞数量显�Objective To investigate the effects of Yishen Zhuyu Tongluo Decoction on chondrocyte pyroptosis and ferroptosis in rats with knee osteoarthritis(KOA).Methods Forty SD were divided randomly into a sham operation group(n=10)and a modeling group(n=30).A KOA model was established in the modeling group by anterior cruciate ligament transection of the right hind limb knee joint.The drawer test was used to confirm the successful establishment of the model.Post-surgery,the rats were subjected to 30 min of forced activity daily to induce KOA.Lameness and hopping movements were observed after 4 weeks,and hematoxylin and eosin staining confirmed cartilage surface damage,deformation,and inflammatory cell infiltration,indicating successful modeling.The model rats were then assigned randomly to a model group(n=8),celecoxib group(n=8),and Yishen Zhuyu Tongluo Decoction group(n=8).The sham operation and model groups received 10 mL/kg of saline by gavage,the celecoxib group received 12 mg/kg of celecoxib solution,and the Yishen Zhuyu Tongluo Decoction group received 5.4 g/kg of the herbal decoction,once daily for 8 weeks.After the interventions,the rats were anesthetized,blood was collected from the abdominal aorta and serum was separated,and the knee joints were isolated.Three samples were fixed in paraformaldehyde,while the remaining cartilage tissue was reserved.Pathological changes in joint cartilage were observed by hematoxylin and eosin and Safranin O-Fast Green staining.Bone microstructure was analyzed using micro-computed tomography.Serum interleukin(IL)-1β and IL-18 protein levels were detected by enzyme-linked immunosorbent assay.Relative mRNA expression levels of NLRP3,ASC,Caspase-1,GSDMD,IL-1β,IL-18,ACSL-4,FTH-1,GPX-4,and cyclooxygenase(COX)-2 were detected by reverse transcription quantitative real-time PCR,and protein expression of NLRP3,Caspase-1,and COX-2 were detected by immunohistochemistry.Results Compared with the sham operation group,model rats showed surface damage and deformation of cartilage tissue,diso

关 键 词：逐瘀通络汤 骨关节炎 细胞焦亡 铁死亡 

分 类 号：R-33[医药卫生]