蒲公英甾醇治疗大鼠膝关节骨关节炎的作用机制初步探索  

Preliminary Study on the Mechanism of Taraxasterol in the Treatment of Knee Osteoarthritis in Rats

作　　者：谢子康[1] 史铭钰 蒋阳[1] 王斌[1] 沈鹏飞[1] 郑冲[1] 杨晓峰[1] Xie Zikang;Shi Mingyu;Jiang Yang;Wang Bin;Shen Pengfei;Zheng Chong;Yang Xiaofeng(Department of Orthopaedics,Changzhou Chinese Medicine Hospital,Chang zhou 213000,China)

机构地区：[1]常州市中医医院骨一科,江苏常州213000

出　　处：《实用骨科杂志》2022年第5期416-421,共6页Journal of Practical Orthopaedics

基　　金：2020年常州市科技局第十六批科技项目(CJ20200109)。

摘　　要：目的初步探索蒲公英甾醇治疗大鼠膝关节骨关节炎(knee osteoarthritis,KOA)的作用机制。方法采用4%木瓜蛋白酶溶液+0.03 mmol/L半胱氨酸混合液0.2 mL关节腔内注射制备KOA大鼠模型,分别给予蒲公英甾醇2.5 mg/kg(低剂量组)、5 mg/kg(中剂量组)、10 mg/kg(高剂量组)及生理盐水(模型组)灌胃,关节腔注射生理盐水0.2 mL制备假手术组,给予生理盐水灌胃;给药4周后,用酶联免疫吸附法测定血清肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素(interleukin,IL)-1β、IL-6、基质金属蛋白酶(matrix metalloproteinase,MMP)-1、MMP-3、半胱氨酸蛋白酶-1(cysteine aspartic acid protease,Capase-1)水平,采用实时荧光定量聚合酶链式反应(real time-quantitative polymerase chain reaction,RT-qPCR)法测定关节软骨组织中微小RNA(microRNA,miRNA)-140、miRNA-146a表达水平,采用Western blot法测定软骨组织中的核因子κB抑制因子(inhibitor of NF-κB,IκB)、IκB激酶β(inhibitorκB kinaseβ,IKKβ)、核因子κB(nuclear factor-κB,NF-κB)、转化生长因子β活化激酶1(transforming growth factor-β-activated kinase 1,TAK1)、NOD样受体家族3(NOD-like receptors 3,NLRP3)、凋亡相关微粒蛋白(apoptosis-associated speck-like protein containing CARD,ASC)表达水平;对关节软骨组织进行病理观察。结果与假手术组比较,高、中、低剂量组及模型组的血清TNF-α、IL-1β、IL-6、MMP-1、MMP-3、Capase-1水平及关节软骨组织中IκB、IKKβ、NF-κB、TAK1、NLRP3、ASC蛋白表达水平、Mankin评分均明显增高(P<0.05),关节软骨组织中miRNA-140、miRNA-146a表达水平均明显降低(P<0.05);与模型组比较,高、中、低剂量组的血清TNF-α、IL-1β、IL-6、MMP-1、MMP-3、Capase-1水平及关节软骨组织中IκB、IKKβ、NF-κB、TAK1、NLRP3、ASC蛋白表达水平、Mankin评分均明显降低(P<0.05),并且随着剂量的增加而明显降低(P<0.05),关节软骨组织中miRNA-140、miRNA-146a表达水平均明显�Objective To study the mechanism of taraxasterol in the treatment of knee osteoarthritis(KOA)in rats.Methods KOA rats model was prepared by injecting 0.2 mL of 4%papain solution+0.03 mmol/L cysteine into the joint cavity.The taraxasterol 2.5 mg/kg(low-dose group),5 mg/kg(medium-dose group),10mg/kg(high-dose group)and saline(model group)were administered by gavage,respectively.The sham operation group was prepared by injecting 0.2 mL of normal saline into the joint cavity and gavage with normal saline.4 weeks after administration,the serum levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,matrix metalloproteinase-1(MMP-1),MMP-3 and cysteine aspartic acid protease-1(Capase-1)in were determined by enzyme-linked immunosorption.The expression levels of microRNA(miRNA)-140 and miRNA-146a in articular cartilage tissue were determined by real-time PCR(RT-qPCR).The protein expression levels of inhibitor of NF-κB(IκB),inhibitorκB kinaseβ(IKKβ),nuclear factor-κB(NF-κB),transforming growth factor-β-activated kinase 1(TAK1),NOD-like receptors 3(NLRP3),apoptosis-associated speck-like protein containing CARD(ASC)in articular cartilage tissue were determined by Western blot.The pathological observation of articular cartilage was conducted.Results Compared with the sham operation group,the serum levels of TNF-α,IL-1β,IL-6,MMP-1,MMP-3,Capase-1 and the protein expression levels of IκB,IKKβ,NF-κB,TAK1,NLRP3,ASC and Mankin score in articular cartilage tissue in high-dose,medium-dose,low-dose group and model group was significantly increased(P<0.05),the expression levels of miRNA-140 and miRNA-146a in articular cartilage were significantly decreased(P<0.05).Compared with the model group,the serum levels of TNF-α,IL-1β,IL-6,MMP-1,MMP-3,Capase-1 and the protein expression levels of IκB,IKKβ,NF-κB,TAK1,NLRP3,ASC and Mankin score in articular cartilage tissue in high-dose,medium-dose and low-dose group was significantly decreased(P<0.05),it decreased significantly with the increase of dose(P<0.05),t

关 键 词：膝关节骨关节炎 大鼠 蒲公英甾醇 炎性因子 信号通路 微小RNA 作用机制 

分 类 号：R684.3[医药卫生—骨科学]