基于TLR4/NF-κB通路探讨梅花入骨丹水提取物改善膝骨关节炎大鼠软骨基质代谢的作用机制  

Mechanism of Bauhinia championii Aqueous Extract to Improve Cartilage Matrix Metabolism in Knee Osteoarthritis Rats Based on TLR4/NF-κB Signaling Pathway

作　　者：林家钟[1] 黄艳峰 刘蔚楠[1] 陈妍[1] 贺浪 陈清 林翔[1] LIN Jiazhong;HUANG Yanfeng;LIU Weinan;CHEN Yan;HE Lang;CHEN Qing;LIN Xiang(The Affiliated People's Hospital of Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350004,China;Academy of Integrative Medicine,College of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China)

机构地区：[1]福建中医药大学附属人民医院,福建福州350004 [2]福建中医药大学中西医结合学院中西医结合研究院,福建福州350122

出　　处：《福建中医药》2024年第12期18-23,33,共7页Fujian Journal of Traditional Chinese Medicine

基　　金：福建省自然科学基金项目(2023J01838);福建省卫生健康科技计划项目(2023GGA070);福建中医药大学医学技术学科专项课题(X202304-医技)。

摘　　要：目的基于TLR4/NF-κB通路探讨梅花入骨丹水提取物(BCAE)改善膝骨关节炎(KOA)大鼠软骨基质代谢失衡的作用机制。方法将42只2月龄SPF级SD大鼠按随机数字表法分为空白组13只和造模组29只。造模组采用关节腔注射木瓜蛋白酶法建立大鼠右膝KOA模型,2组各随机抽取5只通过肉眼和光镜下观察软骨组织切片评分进行模型鉴定。模型鉴定成功后,再将造模组按随机数字表法分为模型组、BCAE组和扶他林组,每组8只。BCAE组按体质量每天1 mL/100 g灌胃0.09 g/mL梅花入骨丹水提取物,扶他林组按体质量每天1.0 mL/100 g灌胃0.675 mg/mL扶他林混悬液,空白组和模型组分别给予等剂量生理盐水灌胃,连续28 d。观察干预后4组大鼠一般情况及体质量、右膝关节直径、右下肢行走足迹及行走疼痛的变化情况。取大鼠膝关节软骨组织,HE染色后在光镜下观察软骨组织病理变化并进行改良Mankin's评分,Western blot检测软骨组织TLR4、MyD88、p-p65/p65、MMP-1、MMP-3、MMP-13、ColⅡ及Aggrecan蛋白表达量。结果与空白组比较,模型组大鼠出现体质量下降,行走疼痛,右膝关节肿胀,软骨退变,Mankin's评分显著增高(P<0.05);与模型组比较,BCAE组和扶他林组大鼠体质量增加,行走疼痛及右膝关节肿胀减轻,软骨退变程度较轻,Mankin's评分均明显降低(P<0.05)。与空白组比较,模型组大鼠膝关节软骨TLR4、MyD88、p-p65/p65、MMP-1、MMP-3及MMP-13蛋白表达量明显增加,ColⅡ及Aggrecan蛋白表达量显著降低(P<0.05);与模型组比较,BCAE组和扶他林组软骨TLR4、MyD88、p-p65/p65、MMP-1、MMP-3及MMP-13蛋白表达量明显降低,而ColⅡ及Aggrecan蛋白表达量显著增加(P<0.05)。结论梅花入骨丹可抑制TLR4/NF-κB信号通路促进KOA大鼠软骨组织MMP-1、MMP-3、MMP-13的蛋白表达量,改善KOA大鼠软骨基质代谢失衡,减轻关节炎症反应,延缓软骨退变。Objective:Based on TLR4/NF-κB pathway,to explore the mechanism of Bauhinia championii aqueous extract(BCAE)in improving the cartilage matrix metabolism in rats with knee osteoarthritis(KOA).Methods:A total of 42 SPF level SD rats aged two months were randomly selected using random number table,with 13 rats in the blank group and the remaining 29 rats in the modeling group.The rat models of KOA were established by intra-articular injection of papain.Five rats were randomly selected from each of the two groups for model verification by visually inspecting and scoring cartilage tissue sections under optical microscope.After successful modeling,the modeling group was further divided into the model group,the BCAE group,and the Voltaren group using a random number table method,with 8 rats in each group.The BCAE group was gavaged daily with 0.09 g/mL Bauhinia championii herbal liquid at a dose of 1 mL/100 g of body weight.The Voltaren group was gavaged daily with 0.675 mg/mL Voltaren suspension at a dose of 1.0 mL/100 g of body weight.The blank group and the model group were gavaged with an equivalent volume of saline,respectively,for 28 consecutive days.The changes of general condition,body weight,diameter of the right knee joint,walking footprints of the right hind limb,and walking pain of the rats in the four groups were observed after intervention.The cartilage tissues of the knee joints were cut to observe the pathological changes under optical microscope and modified Mankin's score were performed.Western blot method was used to detect the protein expression of TLR4,MyD88,p-P65/P65,MMP-1,MMP-3,MMP-13,Col II and Aggrescan in the cartilage.Results:Compared with the blank group,rats in the model group showed loss of body mass,walking pain,swelling of the right rear knee joint,cartilage degeneration,and a significant increase in Mankin's scores(P<0.05).Compared with the model group,rats in the BCAE and Voltaren groups showed an increase in body mass,a decrease in walking pain and swelling of the right rear knee joint,

关 键 词：膝骨关节炎 梅花入骨丹 软骨退变 基质代谢 TLR4/NF-κB信号通路 

分 类 号：R68[医药卫生—骨科学]