曲美他嗪对大鼠骨关节炎炎症反应和软骨基质降解的作用研究  

Effects of trimetazidine on inflammation and cartilage matrix degradation in rats with osteo⁃arthritis

作　　者：胡紫薇 茹晓 蓝逆寒 高明 郑立[1,2,3] Hu Ziwei;Ru Xiao;Lan Nihan;Gao Ming;Zheng Li(Guangxi Research Center of Medical Biomaterials Engineering and Technology for Tissue and Organ Repair,Guangxi Medical University,Nanning 530021,China;Pharmaceutical College,Guangxi Medical University,Nanning 530021,China;Provincial and Ministerial Collaborative Innovation Center for Regenerative Medicine and Medical Biological Resources Development and Application,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学广西组织器官修复医用生物材料工程技术研究中心,南宁530021 [2]广西医科大学药学院,南宁530021 [3]广西医科大学再生医学与医用生物资源开发应用省部共建协同创新中心,南宁530021

出　　处：《广西医科大学学报》2022年第4期513-519,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.82160430);广西科创新驱动发展专项资助项目(No.桂科AA19254002);广西医科大学青年科学基金项目(No.GXMUYSF202111)。

摘　　要：目的:探讨曲美他嗪对大鼠骨关节炎(OA)炎症反应和软骨基质降解作用。方法:将软骨细胞分为对照组、模型组和实验组。对照组加入单纯培养基培养,模型组加入含有10μg/mL IL-1β培养基培养,实验组加入含有10μg/mL IL-1β+0.1 mg/mL曲美他嗪的培养基培养。通过细胞计数试剂(CCK-8)法和钙黄绿素/碘化丙啶(Calcein-AM/PI)染色法检测曲美他嗪的毒性作用;实时荧光定量PCR(RT-qPCR)检测软骨形成和炎症相关基因的表达;将大鼠随机为对照组、模型组和实验组,除对照组外,模型组和实验组采用前交叉韧带切断手术建立大鼠OA模型,实验组大鼠关节腔内注射100μL 0.1 mg/mL曲美他嗪,模型组和对照组分别注射等量的0.9%NaCl溶液。处理4周后,对各组关节进行组织学染色及评分。结果:CCK-8结果显示,与0 mg/mL比较,曲美他嗪浓度低于0.1 mg/mL时,对软骨细胞无明显毒性(P>0.05);Calcein-AM/PI染色结果显示,与模型组比较,实验组活细胞数量增加,死细胞数量减少,细胞活力提高(P<0.01);RT-qPCR结果显示,与模型组比较,实验组的性别决定区域Y相关的高迁移率族框9(SOX9)和蛋白聚糖(ACAN)相对表达量均升高(均P<0.05),白介素-6(IL-6)、基质金属蛋白酶-3(MMP-3)、基质金属蛋白酶-13(MMP-13)、环氧酶-2(COX-2)及肿瘤坏死因子-α(TNF-α)表达均降低(均P<0.05)。相较于模型组,实验组软骨层的损伤与变形明显缓解;与模型组相比,实验组的组织学评分降低(P<0.05)。结论:曲美他嗪具有减轻OA炎症反应和保护软骨的作用。Objective:To investigate the effects of trimetazidine on inflammation and cartilage matrix degradation in rats with osteoarthritis(OA).Methods:Chondrocytes were divided into control group,model group and experimental group.The control group was cultured with simple medium,the model group was cultured with 10μg/mL IL-1βmedium,and the experimental group was cultured with 10μg/mL IL-1β+0.1 mg/mL trimetazidine.The toxicity of trimetazidine was detected by CCK-8 method and Calcein/propidium iodide(Calcein-AM/PI)staining,and the expression of genes related to chondrogenesis and inflammation was detected by RT-qPCR.Rats were randomly divided into control group,model group and experimental group.Except for control group,OA model was established by anterior cruciate ligament transection in model group and experimental group.100μL 0.1 mg/mL trimetazidine was injected into articular cavity of rats in experimental group,and the same amount of 0.9%NaCl solution was injected into model group and control group.After 4 weeks of treatment,the joints of each group were stained and scored.Results:The results of CCK-8 showed that trimetazidine had no obvious toxicity to chondrocytes when the concentration of trimetazidine was lower than 0.1 mg/mL(P>0.05).Compared with model group,Calcein-AM/PI staining showed that the number of living cells increased,the number of dead cells decreased and the cell viability increased in the experimental group(P<0.01).Compared with model group,the mRNA expressions of high mobility group frame 9(SOX9)and proteoglycan(ACAN)related to gender determination region Y in the experimental group were higher than those in the model group(P<0.05).The expressions of interleukin-6(IL-6),matrix metalloproteinase-3(MMP-3),matrix metalloproteinase-13(MMP-13),cyclooxygenase-2(COX-2)and tumor necrosis factor-α(TNF-α)were decreased(P<0.05).Compared with the model group,the injury and deformation of the cartilage layer in the experimental group were alleviated,and the histological score in the experimental group

关 键 词：曲美他嗪 骨关节炎 炎症 软骨细胞外基质 

分 类 号：R961.1[医药卫生—药理学]