摘要
目的观察陈元膏摩法对丙二醛(Malondialdehyde,MDA)、二价铁离子(Ferrousion,Fe^(2+))、活性氧(Reactive oxygen species,ROS)以及白细胞介素-1β(Interleukin-1β,IL-1β)、白细胞介素-6(Interleukin-6,IL-6)以及肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)表达的影响,探究陈元膏摩法治疗膝骨性关节炎的效果及作用机制。方法24只新西兰大白兔随机抽取6只为空白组,其余18只采用前交叉韧带切断术(Anterior cruciate ligament transection model,ACLT)制备膝骨性关节炎(Knee osteoarthritis,KOA)模型,将成模KOA兔随机分为模型组、扶他林组、陈元膏摩法组,每组6只。空白组和模型组不予干预,陈元膏摩法组在涂抹陈元膏基础上推拿治疗,每日1次,连续治疗4周。扶他林组每只兔用扶他林软膏在相同部位1 g涂抹,推拿手法操作同陈元膏摩法组。干预前1 d、干预后28 d,通过奎森功能障碍指数(Lequesne functional severity index,Lequesne MG)评估膝关节。干预结束后次日取材,苏木精-伊红(Hematoxylin and eosin,HE)染色法观察软骨组织病理学变化;透射电镜观察线粒体形态;酶联免疫吸附(Enzyme linked immunosorbent assay,ELISA)试剂盒等检测血清中IL-1β、IL-6、TNF-α、MDA、Fe^(2+)、ROS含量。结果病理结果显示,空白组兔膝关节软骨表面光滑,软骨层及。软骨细胞形态良好,软骨下骨及骨小梁结构规则,滑膜组织未见明显增生,滑膜细胞线粒体的形态结构较正常。模型组兔膝关节软骨结构破坏,部分软骨层缺失伴纤维组织增生、软骨细胞退变、坏死、软骨下骨及骨小梁结构紊乱伴结构破坏,滑膜组织增生伴较多炎细胞浸润,滑膜细胞细胞核核周隙增宽;胞浆中线粒体明显肿胀,体积变大,嵴减少、断裂或消失,基质大量溶解呈絮状甚至空泡状。扶他林组和陈元膏摩法组膝关节软骨结构破坏程度及滑膜细胞线粒体形态结构较模型组均有所改善。干预前,与空白组相比,模型组、扶他林组和陈元膏摩法组兔膝关节Lequesne MG评分升高(P<0.05);干预后,与空白组相比,模型组软骨组织病理损伤严重,膝关节Lequesne MG评分、曼金(Mankin osteoarthritis histological grading system,Mankin′s)评分、血清中IL-1β、IL-6、TNF-α表达升高(P<0.05)。与模型组相比,陈元膏摩法组软骨组织病理损伤有所好转,膝关节Lequesne MG评分、Mankin′s评分、血清中IL-1β、IL-6、TNF-α含量以及滑膜组织中MDA、Fe^(2+)、ROS表达降低(P<0.05)。结论陈元膏摩法可能是通过抑制脂质过氧化,降低炎症因子的表达,进而改善KOA兔软骨损伤及炎症反应。
Objective To observe the effects of Chen Yuan Ointment Rubbing Method on malondialdehyde(MDA),ferrous ion(Fe^(2+)),reactive oxygen species(ROS)and the expression of interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α),and to explore the therapeutic effect and mech-anism of Chen Yuan Ointment Rubbing Method on knee osteoarthritis(KOA).Methods 24 New Zealand white rabbits were randomly divided into a blank group(6 rabbits)and a KOA model group(18 rabbits).The KOA model was established by anterior cruciate ligament transection(ACLT).The KOA rabbits were randomly divided into a model group,a Voltaren group,and a Chen Yuan Ointment Rubbing Method group(6 rabbits each).The blank group and the model group received nointervention.The Chen Yuan Ointment Rubbing Method group was treated with Chen YuanOintment and massage once a day for 4 weeks.The Voltaren group was treated with 1 gof Voltaren ointment on the same area and the massage technique was the same as the Chen Yuan Ointment Rubbing Method group.Lequesne functional severity index(Lequesne MG)was evaluated 1 day before and 28 days after the intervention.The next day after the intervention,the samples were collected.Hematoxylin and eosin(HE)staining was used to observe the pathological changes of cartilage tissue;transmission electron microscopy was used to observe the mor-phology of mitochondria;Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of IL-1β,IL-6 and TNF-αin serum;and kits were used to detect the levels of MDA,Fe^(2+)and ROS.Results Patholog-ical results showed that the knee joint cartilage surface of blank group rabbits was smooth,thecartilage layer and chondrocytes were in good shape,the subchondral bone and trabecularbone structure was regu-lar,and no obvious hyperplasia of the synovial tissue was observed.The morphology and structure of the mitochondria in the synovial cells were normal.Theknee joint cartilage structure of the model group rabbits were damaged,with partial cartilage layer loss,fibrous tissue hyperplasia,chondrocyte degeneration and necrosis,disorderedand damaged subchondral bone and trabecular bone structure,and synovial tissue hy-perplasiawith a large number of inflammatory cell infiltrations.The nuclear perinuclear space ofthesynovi-al cells was widened;the mitochondria in thecytoplasm were significantly swollen,with increased volume,reduced and broken or disappeared cristae,and the matrixwas dissolved in a flocculent or even vacuolar form.The degree of knee joint cartilage structure damage and the morphology and structure of synovial cell mitochondria in the Voltarengroupand the Chen Yuan Ointment Rubbing Method groupwere improved comparedwith the model group.Before the intervention,the Lequesne MG score of the knee joint in the-model group,Voltaren group,and Chen Yuan Ointment Rubbing Method group was higherthan that of the blank group(P<0.05).After the intervention,the pathological damage ofthe cartilage tissue in the model group was severe,and theLequesne MG score,Mankin osteoarthritis histological grading system(Mankin's)score,and the expression of IL-1β,IL-6 and TNF-αin serum were higher than those of the blank group(P<0.05).Compared withmodel group,the pathological damage of the cartilage tissue in the Chen Yuan Ointment Rubbing Method group was improved,and the Lequesne MG score,Mankin's score,the expressionof IL-1β,IL-6 and TNF-αin serum,and the expression of MDA,Fe^(2+)and ROS in synovial tissu were decreased(P<0.05).Conclusion Chen Yuan Ointment Rubbing Methodmay improvethe car-tilage damage and inflammatory response in KOA rabbits by inhibiting lipid peroxidationand reducingthe expression of inflammatory factors.
作者
王月媛
古来撒尔·艾克拜尔
刘俊昌
牙库甫·亚森
张鑫翔
蒋祥龙
侯运运
WANG Yueyuan;Gulaisaer Aikebaier;LIU Junchang;Yakufu Yasen;ZHANG Xinxiang;JIANG Xianglong;HOU Yunyun(College of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830017,China;Fourth Clinical Medical College of Xinjiang Medical University,Urumqi 830000,China;Xinjiang Uygur Autonomous Region Institute of Traditional Chinese Medicine,Urumqi 830000,China)
出处
《新疆医科大学学报》
2025年第7期881-887,共7页
Journal of Xinjiang Medical University
基金
国家自然科学基金项目(82360979)
新疆维吾尔自治区科技创新团队(天山创新团队)项目(2022TSYCTD0008)
新疆医科大学研究生科研创新项目(CXCY2025019)。
关键词
陈元膏
推拿疗法
膝骨性关节炎
脂质过氧化
炎症因子
Chen Yuan Ointment
massage therapy
knee osteoarthritis
lipid peroxidation
inflammatory factors
