抑制EGFR信号通路对骨折愈合早期骨内、外膜来源干细胞增殖、分化能力影响的研究  

Effect of inhibiting EGFR signaling pathway on the proliferation and differentiation of stem cells derived from bone and adventitia in the early stage of fracture healing

作　　者：王华松 兰生辉 庞炯宇 丰瑞兵 杨晨曦 袁功武 曾文波 Wang Huasong;Lan Shenghui;Pang Jiongyu;Feng Ruibing;Yang Chenxi;Yuan Gongwu;Zeng Wenbo(General Hospital of Central Theater Command,WuhanHubci,430065;Shanghai Eighth People's Hospital,Shanghai,200235;Hubci University of Traditional Chinese Medicine,Wuhan Hubei,430065;Hubei Provincial Hospital of Integrated Chinese and Western Medicine,Wuhan Hubeif 430015;The Third Affiliated Hospital of Chongqing Medical University,Chongqing,401120,China)

机构地区：[1]中国人民解放军中部战区总医院,湖北武汉430065 [2]上海第八人民医院,上海200235 [3]湖北中医药大学,湖北武汉430065 [4]湖北省中西医结合医院,湖北武汉430015 [5]重庆医科大学附属第三医院,重庆401120

出　　处：《生物骨科材料与临床研究》2020年第6期5-8,14,共5页Orthopaedic Biomechanics Materials and Clinical Study

基　　金：湖北省卫生健康委科研联合项目(WJ2019H097)。

摘　　要：目的探索抑制表皮生长因子受体(EGFR)信号通路对骨折愈合早期骨内、外膜来源SCs增殖、分化能力的影响。方法将建立右股骨骨折模型成功的42只SD大鼠按随机数字表分为实验组与对照组,实验组给予Gefitinib 100 mg/(kg·d)(溶于0.5%甲基纤维素)灌胃,对照组给予等量0.5%甲基纤维素灌胃。术后1周提取右股骨骨内、外膜区域的SCs并进行体外培养。通过流式细胞术鉴定第3代干细胞表面标志物FITC-CD29、FITCCD34、FITC-CD45、FITC-CD90;BrdU法检测各组干细胞增殖能力;成骨诱导及成软骨诱导分化后行von Kossa染色及阿尔新蓝染色,检测干细胞的分化能力。结果各组SCs表面标志物FITC-CD29、FITC-CD90呈高表达,FITC-CD34、FITC-CD45呈低表达;BrdU法检测发现实验组骨内、外膜SCs增殖能力较对照组显著降低(P<0.01);成骨诱导分化并染色后,实验组骨内、外膜SCs的矿化结节较对照组更多,成软骨诱导分化并染色后,实验组SCs的淡蓝色颗粒较对照组更多。结论在骨折愈合早期,抑制EGFR信号通路能抑制骨内、外膜来源SCs的增殖,促进其成骨、成软骨的分化。Objective To explore the effect of inhibiting EGFR signaling pathway on the proliferation and differentiation of SCs derived from bone and adventitia in the early stage of fracture healing. Methods Forty-two SD rats successfully established right femur fracture model were randomly divided into experimental group and control group. The experimental group was given Gefitinib 100 mg/(kg·d) by gavage(dissolved in 0.5% methyl cellulose), and the control group was given the same amount of 0.5% methyl cellulose. One week after surgery, the SCs in the inner and outer adventitia of the right femur were extracted and cultured in vitro. The third-generation stem cell surface markers FITCCD29, FITC-CD34, FITC-CD45, FITC-CD90 were identified by flow cytometry;BrdU method was used to detect stem cell proliferation capacity. After osteogenic and chondrogenic differentiation, von Kossa staining and Alcian blue staining were performed to detect the differentiation ability of stem cells. Results The SCs surface markers FITC-CD29 and FITC-CD90 of each group showed high expression, while FITC-CD34 and FITC-CD45 showed low expression;BrdU method detected that the proliferation ability of SCs in the experimental group was significantly lower than that in the control group(P<0.01). After osteogenic differentiation and staining, the experimental group had more mineralized nodules from the inner and outer membranes of the SCs than the control group. After cartilage differentiation and staining,the experimental group had more light blue particles than the control group. Conclusion In the early stage of fracture healing, inhibition of the EGFR signaling pathway can inhibit the proliferation of SCs derived from the inner and outer membranes, and promote the differentiation of osteoblasts and cartilage.

关 键 词：表皮生长因子受体 信号通路 骨膜干细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]