IL-17A通过抑制基质金属蛋白酶的表达降低Masquelet诱导膜的生物活性  

IL-17A reduces the biological activity of Masquelet-induced membranes by inhibiting the expression of matrix metalloproteinases

作　　者：汪凯 毛栋 潘筱云 张元澍 金叶盛 芮永军[1] Wang Kai;Mao Dong;Pan Xiaoyun;Zhang Yuanshu;Jin Yesheng;Rui Yongjun(Department of Orthopedics,Wuxi Ninth Affiliated Hospital of Suzhou University,Wuxi 214062,China)

机构地区：[1]苏州大学附属无锡九院骨科,无锡214062

出　　处：《中华手外科杂志》2024年第1期90-96,共7页Chinese Journal of Hand Surgery

基　　金：无锡市"太湖人才计划"顶尖医疗专家团队项目;无锡市卫生健康委员会科研资助项目(2020ZHYB12);江苏省卫生健康委员会科研资助项目(Z2020010);无锡市科技局创新创业项目(Y20212055)

摘　　要：目的探究Masquelet诱导膜血管生成减少、生物活性降低的信号机制。方法采用克氏针内固定的方法建立大鼠股骨临界骨缺损模型,造模成功后分别于术后2、4、6、8周取出诱导膜样本,用于生物学检测和蛋白组学分析;免疫组化检测诱导膜CD31^(+)血管数量;GO富集分析诱导膜活性降低的生物学过程;KEGG富集分析诱导膜活性降低的信号通路;Western bot检测诱导膜IL-7A、MMP8、MMP9、MMP13的蛋白表达水平。结果CD31免疫组织化学显示,2、4周诱导膜血管生成较多,生物活性较好;4周以后血管逐渐减少,生物活性降低。GO富集分析与诱导膜生物活性降低有关的胶原分解代谢过程,该生物学过程包含基质金属蛋白酶家族的蛋白:MMP8、MMP9、MMP13。KEGG富集分析显示,MMP9、MMP13在IL-17A信号通路中,是IL-17A的下游调节因子,并且IL-17A信号通路在诱导膜活性降低的top20通路中。Western Blot结果验证IL-17A、MMP8、MMP9、MMP13在不同时间点的诱导膜中均表达,8周相较于4周的降低差异有统计学意义。结论IL-17A信号通路及其下游MMP8、MMP9、MMP13的表达降低是诱导膜血管生成减少和生物活性降低的关键因素。Objective To explore the signaling mechanism of Masquelet-induced reduction in membrane angiogenesis and decreased biological activity.Methods A rat femoral critical bone defect model was established using Kirschner wire internal fixation.After successful modeling,induced membrane samples were taken out at 2,4,6,and 8 weeks postoperatively for biological detection and proteomic analysis.Immunohistochemical methods were used to detect the number of induced membrane CD31+blood vessels.GO enrichment was used to analyze the biological process of induced membrane activity reduction.KEGG enrichment was used to analyze the signaling pathways of induced membrane activity reduction.Western blot was used to detect the protein expression levels of induced membrane IL-7A,MMP8,MMP9,and MMP13.Results CD31 immunohistochemistry showed that at 2 and 4 weeks,there was more induced membrane angiogenesis and better biological activity.After 4 weeks,blood vessels gradually decrease and biological activity decreases.GO enrichment analyzed the collagen degradation metabolism process which was related to decreased induced membrane biological activity,which includes proteins of the matrix metalloproteinase family:MMP8,MMP9,MMP13.KEGG enrichment analysis showed that MMP9 and MMP13 were downstream regulatory factors of IL-17A in the IL-17A signaling pathway,and the IL-17A signaling pathway was in the top 20 pathway that decreased induced membrane activity.Western blot results confirmed that IL-17A,MMP8,MMP9,and MMP13 were all expressed in the induced membrane at different time points,and the decrease at 8 weeks was statistically significant compared to 4 weeks.Conclusion The decreased expression of IL-17A signaling pathway and its downstream MMP8,MMP9,and MMP13 is a key factor in induced membrane decreased angiogenesis and biological activity.

关 键 词：动物实验 大鼠 Masquelet技术 诱导膜 基质金属蛋白酶 

分 类 号：R683[医药卫生—骨科学] R-332[医药卫生—外科学]