中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
Chinese Journal of Rheumatology
2015年
11期
756-760,封3
,共6页
骨关节炎%Wnt信号通路%代谢
骨關節炎%Wnt信號通路%代謝
골관절염%Wnt신호통로%대사
Osteoarthritis%Wnt signaling pathway%Metabolism
目的 研究塞来昔布对兔OA模型膝关节软骨细胞基质代谢的作用,并探讨Wnt/β-连环蛋白(β-catenin)信号通路在其中可能发挥的作用.方法 采用改良伸直位固定法制作兔OA模型,分离提取OA兔膝关节软骨细胞进行培养.采用四甲基偶氮唑盐(MTT)比色法选择塞来昔布和二甲基亚砜(DMSO)实验浓度.将软骨细胞分为DMSO对照组和塞来昔布组,分别给予1％DMSO和10 μmol/L塞来昔布干预48 h后,检测软骨细胞Wnt/β-catenin相关因子mRNA及蛋白水平:包括β-catenin、MMP-3、MMP-13、Ⅱ型胶原和蛋白聚糖.2组间比较采用t检验进行统计分析.结果 与对照组相比,塞来昔布组Ⅱ型胶原[(275±19) ng/ml与(202±12) ng/ml,t=-6.21,P＜0.05]和蛋白聚糖[(58±12) ng/ml与(30±3)ng/ml,t=-9.23,P＜0.05]水平均明显升高,差异有统计学意义(P＜0.05).与对照组相比,塞来昔布组β-catenin、MMP-3、MMP-13的基因及蛋白表达水平均明显降低,差异有统计学意义(P＜0.05).结论 塞来昔布10 μmol/L可增加兔OA软骨细胞Ⅱ型胶原和蛋白聚糖含量,其作用机制可能是通过抑制细胞内Wnt/β-catenin信号通路、降低下游因子MMPs表达、减少MMPs对软骨基质Ⅱ型胶原和蛋白聚糖的分解作用完成的.
目的 研究塞來昔佈對兔OA模型膝關節軟骨細胞基質代謝的作用,併探討Wnt/β-連環蛋白(β-catenin)信號通路在其中可能髮揮的作用.方法 採用改良伸直位固定法製作兔OA模型,分離提取OA兔膝關節軟骨細胞進行培養.採用四甲基偶氮唑鹽(MTT)比色法選擇塞來昔佈和二甲基亞砜(DMSO)實驗濃度.將軟骨細胞分為DMSO對照組和塞來昔佈組,分彆給予1％DMSO和10 μmol/L塞來昔佈榦預48 h後,檢測軟骨細胞Wnt/β-catenin相關因子mRNA及蛋白水平:包括β-catenin、MMP-3、MMP-13、Ⅱ型膠原和蛋白聚糖.2組間比較採用t檢驗進行統計分析.結果 與對照組相比,塞來昔佈組Ⅱ型膠原[(275±19) ng/ml與(202±12) ng/ml,t=-6.21,P＜0.05]和蛋白聚糖[(58±12) ng/ml與(30±3)ng/ml,t=-9.23,P＜0.05]水平均明顯升高,差異有統計學意義(P＜0.05).與對照組相比,塞來昔佈組β-catenin、MMP-3、MMP-13的基因及蛋白錶達水平均明顯降低,差異有統計學意義(P＜0.05).結論 塞來昔佈10 μmol/L可增加兔OA軟骨細胞Ⅱ型膠原和蛋白聚糖含量,其作用機製可能是通過抑製細胞內Wnt/β-catenin信號通路、降低下遊因子MMPs錶達、減少MMPs對軟骨基質Ⅱ型膠原和蛋白聚糖的分解作用完成的.
목적 연구새래석포대토OA모형슬관절연골세포기질대사적작용,병탐토Wnt/β-련배단백(β-catenin)신호통로재기중가능발휘적작용.방법 채용개량신직위고정법제작토OA모형,분리제취OA토슬관절연골세포진행배양.채용사갑기우담서염(MTT)비색법선택새래석포화이갑기아풍(DMSO)실험농도.장연골세포분위DMSO대조조화새래석포조,분별급여1％DMSO화10 μmol/L새래석포간예48 h후,검측연골세포Wnt/β-catenin상관인자mRNA급단백수평:포괄β-catenin、MMP-3、MMP-13、Ⅱ형효원화단백취당.2조간비교채용t검험진행통계분석.결과 여대조조상비,새래석포조Ⅱ형효원[(275±19) ng/ml여(202±12) ng/ml,t=-6.21,P＜0.05]화단백취당[(58±12) ng/ml여(30±3)ng/ml,t=-9.23,P＜0.05]수평균명현승고,차이유통계학의의(P＜0.05).여대조조상비,새래석포조β-catenin、MMP-3、MMP-13적기인급단백표체수평균명현강저,차이유통계학의의(P＜0.05).결론 새래석포10 μmol/L가증가토OA연골세포Ⅱ형효원화단백취당함량,기작용궤제가능시통과억제세포내Wnt/β-catenin신호통로、강저하유인자MMPs표체、감소MMPs대연골기질Ⅱ형효원화단백취당적분해작용완성적.
Objective To study effect of celecoxib on Wnt/β-catenin signaling pathway and matrix metabolism of chondrocytes in osteoarthritis (OA).Methods Use a modified way of mechanical fixation to make rabbit OA model.Knee OA separated and extractd rabbit chondrocytes were and then culture in medium.The cultured chondroctyes were divided into the Dimethyl sulfoxide (DMSO) control group and the celecoxib group, which were given 1％ DMSO and 10 μmol/L celecoxib respectively.After 48 hours' intervention, the protein expression and gene expression of Wnt/β-catenin signaling pathway related factors, including β-catenin,matrix metalloproteinase (MMP)-3, MMP-13, collagen Ⅱ were detected.T-test was used for the comparison between the two groups.Results Compared with the control group, the levels of type Ⅱ collagen [(275±19) ng/ml vs (202±12) ng/ml, t=-6.21, P＜0.05] and proteoglycan [(58±12) ng/ml vs (30±3) ng/ml, t=-9.23, P＜0.05] significantly increased in celecoxib group (P＜0.05).Meanwhile, compared with the control group, the levels of β-catenin, MMP-3, MMP-13 mRNA and protein in celecoxib group were significantly reduced (P＜0.05).Conclusion In rabbit OA chondrocytes, celecoxib can inhibit Wnt/β-catenin signaling pathway, reduce downstream factors expression including MMPs, thereby increase the cartilage matrix type Ⅱ collagen and proteoglycan levels.
