摘要
目的探索Notch信号通路对骨关节炎(OA)软骨细胞基质合成的影响及可能的作用机制。方法选择2017年我院收治的8例因膝关节OA行膝关节置换患者的股骨髁软骨作为观察组,再选择1例因外伤行膝上截肢的正常股骨髁软骨作为对照组。所取的标本行组织学检测或细胞分离培养及检测。细胞培养分组按照正常软骨细胞、OA软骨细胞、OA软骨细胞+γ-分泌酶抑制剂DAPT(DAPT组)、OA软骨细胞+重组人Delta4蛋白(DLL4组)。免疫组化定性分析以及Western blot蛋白质印迹技术定量分析Notch-1、Notch-3、Jagged-1、Jagged-2、HES5的表达。结果 Notch信号通路表达水平在OA中发生变化,软骨细胞表型改变,Notch-1、Jagged-1、Jagged-2、HES5表达被激活。γ-分泌酶抑制剂DAPT(20μmol/L)干预Notch信号通路后,抑制OA软骨细胞中Noctch-1、HES5、Jagged-1、Notch-3的表达,对Jagged-2的表达作用不明显。重组人Delta4蛋白(100 ng/μL)干预Notch信号通路后,促进Notch-1、Jagged-2、HES5的表达,对Jagged-1、Notch-3作用不太明显。结论 Notch信号通路表达水平在OA中发生变化;DLL4能激活Notch信号通路,DAPT能抑制Notch信号通路。
Objective To explore the role of Notch signaling pathway in the matrix synthesis of osteoarthritis cartilage cell and its possible mechanism.Methods Selected the femoral condylar cartilage of 8 patients who were admitted into our hospital and treated with knee joint replacement as the observation group,while the normal femoral condyle cartilage of one patient with above-knee amputation was selected as the control group.The specimens were given histological examination or cell isolation culture and detection.The cultured cells were divided into 4 groups,namely the normal cartilage cells,OA cartilage cells,OA cartilage cells withγ-secretase inhibitor DAPT,OA cartilage cells with recombinant human proteins Delta4.Then immunohistochemistry and Western blot were used for detecting the expression of Notch-1,Notch-3,Jagged-1,Jagged-2 and HES5 in chondrocytes in vitro.Results The expression of Notch signaling pathway and the phenotype of cartilage cells changed in the osteoarthritis.The expression of Notch-1,Jagged-1,Jagged-2 and HES5 were activated.γ-secretase inhibitor DAPT(20μmol/L)could inhibit the expression of Notch-1,HES5,Jagged-1,and Notch-3,while it had no obvious effect on the expreesion of Jagged-2.Recombinant human proteins Delta4(100 ng/μL)could promote the expression of Notch-1,Jagged-2,and HES5,and it had no obvious effect on the expreesion of Jagged-1 and Notch-3.Conclusion The expression of Notch signaling pathway of cartilage cells changed in the osteoarthritis.DLL4 can activate the Notch signaling pathway and DAPT can inhibit the Notch signaling pathway.
作者
张清
向明
陈杭
杨国勇
胡晓川
李一平
潘飞旭
邓明月
ZHANG Qing;XIANG Ming;CHEN Hang;YANG Guo-yong;HU Xiao-chuan;LI Yi-ping;PAN Fei-xu;DENG Ming-yue(Department of Upper Limb,Sichuan Orthpaedics Hospital,Chengdu Sichuan 610041,China)
出处
《局解手术学杂志》
2018年第4期240-245,共6页
Journal of Regional Anatomy and Operative Surgery
