糖原合成酶激酶-3β信号对炎性环境骨髓间充质干细胞成软骨分化的作用  

Effect of glycogen synthase kinase -3β in chondrogenic differentiation of bone mesenchymal stem cells in inflammatory conditions

作　　者：程鹏 任晔 吴颖星 张津铭[1] 黄鑫 陈安民[1] 杨卿 Cheng Peng;Ren Ye;Wu Yingxing;Zhang Jinming;Huang Xin;Chen Anmin;Yang Qing(Department of Orthopaedic Surgery,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,武汉430030

出　　处：《中华实验外科杂志》2018年第11期1995-1998,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81601951）;中央高校基本科研业务费专项（2014QN103）

摘　　要：目的探讨糖原合成酶激酶-3β（GSK-3β）信号对白细胞介素-1β（IL-1β）介导的炎性环境下骨髓间充质干细胞（BMSCs）成软骨分化过程的作用和机制。方法以IL-1β刺激诱导炎性环境，同时予以10nmol／LGSK-3β抑制剂氯化锂（LiCl）或10nmol／LGSK-3β干预BMSCs成软骨分化过程，定量检测糖胺聚糖（GAG）含量，实时定量反转录聚合酶链反应（RT-qPCR）检测成软骨相关基因性别决定区Y框蛋白9（Sox 9）、Ⅱ型胶原（Collagen 2a）、蛋白多糖（Aggrecan）的mRNA水平，Western blot检测炎症通路总体核转录因子-κB（NF-κB）和磷酸化NF-κB（p-NF-kB）蛋白表达，胞质及胞核中NF-kB蛋白表达。结果IL-1β组的BMSCs成软骨分化指标分泌型GAG含量[（0．806±0．102）μg／ml]以及Sox 9、Collagen 2a、Aggrecan的mRNA水平较空白对照组明显减少（P=0．021、0．039、0．017、0．025）；而IL-1β刺激诱导炎性环境后，LiCl组成软骨分化指标GAG含量[（0．696±0．095）μg／ml]以及Sox 9、Collagen 2a、Aggrecan的mRNA水平较IL-1β组明显增高（P=0．029、0．031、0．014、0．018），NF-kB蛋白稍增多，p-NF-κB却显著减低（P=0．008），胞核NF-kB显著减低（P=0．031），而GSK-3β组的成软骨分化指标及NF-kB蛋白表达情形均与LiCl组相反。结论抑制GSK-3β信号能减弱NF-κB蛋白的磷酸化水平和入核能力，从而抑制NF-kB通路，进而增强BMSCs在IL-1β诱导炎性环境下的成软骨能力。Objective To investigate the effect of Glycogen synthase kinase -3β（GSK-3β） inhibitor in chondrogenic differentiation of bone mesenchymal stem cells （BMSCs） in inflammatory conditions induced by interleukin-1β （ IL-1β ） and possible mechanism. Methods BMSCs were treated with IL-1β in the process of chondrogenic differentiation. Upon with IL-1β, one group was treated with 10 nmol/L LiCl and the other group with 10 nmol/L GSK-3β. For each group, the glycosaminoglycan （GAG） amount was quantitatively tested, and the mRNA of Sox 9, Collagen 2a, Aggrecan were tested by real-time quantitative polymerase chain reaction （ RT-qPCR）. The total nuclear factor-κB（NF-κB） protein, p-NF-κB protein and the NF-κB protein in cytosol or nucleus were tested by Western blotting. Results The index of chondrogenesis, such as the mRNA of Sox 9, Collagen IIa, Aggrecan and the amount of GAG, were significantly decreased in IL-1β group （P=0.021,0.039,0.017,0.023 ）. However, although in inflammatory conditions induced by IL-1β, the indexes of chondrogenesis in LiCl group were significantly increased （P=0.029,0. 031,0. 014,0. 018 ）. Additionally, total NF-κB protein was increased, p-NF-κB protein was significantly decreased （ P=0.008）, and NF-κB protein in nucleus was significantly decreased （P=0.031）. All of these in GSK-3β group were just opposite to LiCl group. Conclusion The ability of BMSCs chondrogenic differentiation were decreased in inflammatory conditions induced by IL-1β, but GSK-3β inhibitor could enhance the ability although with IL-1β. The possible mechanism was GSK-3β inhibitor promotes the phosphorylatiou of NF-κB protein and the transfer into nucleus, and then suppresses NF-κB signal pathway.

关 键 词：糖原合成酶激酶-3Β 核因子-κB通路 炎性环境 成软骨分化 

分 类 号：R68[医药卫生—骨科学]