摘要
基于大鼠关节疼痛、软骨病理程度、ECM降解过程和炎症介质生成水平,探究银杏内酯C(GC)对2种造模方式的软骨保护效果.选取25只Sprague-Dawley大鼠随机分为5组:对照组(Control组)、模型1组(ACLT组)、给药1组(ACLT+GC组)、模型2组(MIA组)和给药2组(MIA+GC组).试验4周后,收集大鼠右后肢股骨、胫骨和血液样本,应用番红O固绿染色和OARSI评分对大鼠股骨和胫骨病理程度进行评估;免疫组化检测软骨中CollagenⅡ和MMP-13 的表达水平;ELISA 检测大鼠血清中 MMP-3、MMP-13、CTX-Ⅱ、COMP、COX-2、INOS、IL-1β 和 TNF-α 的水平变化;冷敏感性试验和伸膝发声试验检测大鼠关节疼痛程度.结果显示,与MIA组相比,ACLT可造成更严重关节软骨结构损伤.ACLT组中股骨和胫骨的OARSI评分、MMP-13的表达和血清中MMP-13、MMP-3、CTX-Ⅱ、COMP水平均高于MIA组;然而ACLT组中炎症介质COX-2、IL-1β和TNF-α水平显著低于MIA组(P＜0.01).GC干预后可降低OARSI评分(P＜0.05或P＜0.01)和疼痛评分,抑制ECM基质降解酶(MMP-13、MMP-3)、软骨代谢标志物(CTX-Ⅱ、COMP)和炎症介质(COX-2、INOS、IL-1β和TNF-α)的表达,并促进Collagen Ⅱ的合成.结果表明,2种造模方式均可造成软骨损伤,其中ACLT+PMMx法构建OA模型中大鼠关节损伤明显,有利于研究软骨结构损伤程度.GC干预后可减轻2组大鼠OA模型中关节疼痛、软骨病理变化、基质降解程度和炎症反应,从而发挥保护软骨作用.
Abstract
The cartilage-protective effect of ginkgolide C(GC)on the two modeling modalities was investigated based on joint pain,degree of cartilage pathology,ECM degradation process,and level of inflammatory mediator production in rats.Twenty-five SD rats were selected and randomly di-vided into five groups:the control group(Control group),model 1 group(ACLT group),adminis-tration 1 group(ACLT+GC group),model 2 group(MIA group),and administration 2 group(MIA+GC group.)The rats were euthanized after 4 weeks of the test.Femur,tibia and blood samples were collected from the right hind limb of rats.The degree of pathology in the femur and tibia of rats was assessed by saffron O solid green staining and OARSI score.Immunohistochemis-try was used to detect the expression levels of collagen Ⅱ and MMP-13 in cartilage.ELISA was used to detect the changes in the levels of MMP-3,MMP-13,CTX-Ⅱ,COMP,COX-2,INOS,IL-1β,and TNF-α in the serum of rats.Cold sensitivity test and knee extension vocalization test were conducted to detect the degree of joint pain in rats.ACLT could cause more severe structural dam-age to articular cartilage compared with the MIA group.The OARSI scores and the expression of MMP-13 in femur and tibia,and the serum levels of MMP-13,MMP-3,CTX-Ⅱ,and COMP were higher in the ACLT group than those in the MIA group.However,the levels of inflammatory me-diators COX-2,IL-1β,and TNF-α were significantly lower in the ACLT group than in the MIA group(P＜0.0l).GC intervention reduced the OARSI score(P＜0.05 or P＜0.01)and pain scores,inhibited the ECM matrix degrading enzymes(MMP-13,MMP-3),cartilage metabolism markers(CTX-11,COMP),and inflammatory mediators(COX-2,INOS,IL-1β and TNF-α)ex-pression,and promoted collagen Ⅱ synthesis.Both modeling methods resulted in cartilage damage.In particular,the OA model constructed by ACLT+PMMx method in rats had obvious joint dam-age,which was favorable to investigate the degree of cartilage structural damage.GC attenuated cartilage pathological changes,pain severity and inflammatory response in the rat OA model in both groups,thus exerting a cartilage-protective effect.
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