褪黑素对骨性关节炎大鼠关节软骨中TGF-β_1和IL-1β表达的影响  

Effects of melatonin on the expression of TGF-β_1 and IL-1β in articular cartilage of rat knee with osteoarthritis

作　　者：黄冲[1] 刘金钊[1] 胡有谷[1] 

机构地区：[1]青岛大学医学院附属医院关节外科,青岛266003

出　　处：《中国矫形外科杂志》2010年第19期1635-1639,共5页Orthopedic Journal of China

摘　　要：[目的]探讨褪黑素对骨性关节炎(osteoarthritis,OA)雄性大鼠关节软骨的影响。[方法]雄性SD大鼠40只,通过在大鼠膝关节腔注射4%papain(木瓜蛋白酶)溶液0.2 ml制作骨性关节炎模型和采用持续光照的方法建立大鼠去松果体模型。实验随机分为4组:Ⅰ组:正常对照组(n=10);Ⅱ组:骨性关节炎组(n=10);Ⅲ组:Ⅱ组+持续24 h光照组(n=10);Ⅳ组:Ⅲ组+给予褪黑素治疗(n=10)。成功建立模型1周后,Ⅳ组左膝关节腔注射浓度为20 mg/ml褪黑素溶液0.2 ml,每周4次,共4周。注射最后1次后,应用ELISA测取大鼠2 Am和2 Pm的血清褪黑素含量,并处死全部老鼠,同时切取股骨髁进行大体观察,之后脱钙制片,行HE染色观察和IL-1β、TGF-β1免疫组化观察(均采用Mankin法进行评分)。[结果](1)大体观察:Ⅰ组软骨表面光滑,有弹性且边缘规整;Ⅱ组软骨表面凹凸不平,失去原有的光泽,存在软骨软化现象及软骨下骨外露;Ⅲ组较Ⅱ组严重;Ⅳ组软骨软化现象消失及软骨剥脱,软骨下骨外露减少。(2)组织学观察:Ⅰ组软骨细胞大小一致,排列规整;Ⅱ组软骨细胞排列紊乱,局部可见簇聚现象;Ⅲ组较Ⅱ组严重;Ⅳ组软骨细胞排列较规则,炎性细胞侵润明显较少。(3)免疫组化观察:Ⅳ组关节软骨IL-1β表达显著低于Ⅲ组和Ⅱ组。Ⅳ组关节软骨TGF-β1表达显著高于Ⅲ组和Ⅱ组。同时,4组组织学和免疫组化的Mankin评分比较差异均有统计学意义。[结论]雄性OA模型大鼠在持续光照下可以加快骨性关节炎关节软骨的的退变进程。浓度为20 mg/ml的褪黑素溶液0.2 ml关节腔注射4周后能够抑制骨性关节炎的进一步发展,其可能作用机制与上调软骨生长因子TGF-β1及下调炎性因子IL-1β表达有关。[Objective]The study was to evaluate the effects of melatonin on the articular cartilage of osteoarthritis（OA） in male rats. [Methods]Forty male rats were randomly divided into four groups.In groupⅠ： ten rats without treatment served as control group.In groupⅡ,Ⅲ and Ⅳ,knee joint of ten rats in each group received intra-articular injection 0.2 ml of 4% papain solution for establishing an OA model.Meanwhile,the rats underwent constant intensive light condition for establishing pinealectomy model in group Ⅲ.Pinealectomy model and melatonin treatment were given in group Ⅳ.One week after OA model was successfully created,the rats of group Ⅳwere treated with intra-articular injections 0.2 ml of 20 mg/ml melatonin solution four times a week for four weeks.One week after the last melatonin injection,the level of serum melatonin in 2 Am and 2 Pm was detected by ELASA.All rats were sacrificed to collect cartilage of femoral condyle for gross observation.After decalcification and paraffin sectioning,pathological analysis was done with HE staining and evaluated by Mankin scores.Expression levels of IL-1β,TGF-β1 were detected with immunostaining and graded according to Mankin scores.[Results]（1） The gross observation showed that articular cartilage surface was smooth,flexible and regular edge in groupⅠ.Cartilage surface was uneven,lost their luster,and had softened cartilage and subchondral bone exposure in group Ⅱ,and even more severe in group Ⅲ.Softened cartilage disappeared,subchondral bone exposure and cartilage denudation were decreased in group Ⅳ.（2）The histological observation showed that articular chondrocytes had the same sizes and arranged regularly in group Ⅰ.The chondrocytes arranged irregularly and cluster formation could be seen at some local area in groupⅡ,and even more severe group Ⅲ.In group Ⅳ,there were less inflammatory infiltrated cytokine,with regular chondrocyte arrangement.（3） Observation of immunohistochemistry： In group Ⅳ,expression of IL-1β was s

关 键 词：褪黑素 骨性关节炎 去松果体 关节软骨 白介素-1Β 转化生长因子-β1 

分 类 号：R684.7[医药卫生—骨科学]