猪脱细胞真皮基质与肿瘤坏死因子-α调控毛囊角质细胞Wnt3a/β-catenin信号通路表达的实验研究  

The role of porcine acellular dermal matrix and tumor necrosis factor-α in regulating the expression of Wnt3a/β-catenin signaling pathway in hair follicle keratinocytes: an experimental study

作　　者：张文文 李恭驰[2] 邹利军[1] 冯自波[1] 杜烨 胡映月 邓海波 祝友鹏 潘银根[3] 李炳辉[1] Zhang Wenwen;Li Gongchi;Zou Lijun;Feng Zibo;Du Ye;Hu Yingyue;Deng Haibo;Zhu Youpeng;Pan Yingen;Li Binghui(Department of Wound Repair,Liyuan Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei 430077,China;Department of Hand Surgery,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei 430022,China;Department of Plastic Surgery,People's Hospital of Qidong City,Qidong,Jiangsu 226200,China)

机构地区：[1]华中科技大学同济医学院附属梨园医院创面修复科,湖北武汉430077 [2]华中科技大学同济医学院附属协和医院手外科,湖北武汉430022 [3]江苏省启东市人民医院整形美容科,江苏启东226200

出　　处：《感染．炎症．修复》2020年第3期162-166,F0002,共6页Infection Inflammation Repair

基　　金：国家自然科学基金青年科学基金项目(81801922);国家卫生健康委员会和财政部资助项目(2019542);湖北省慢性创面及糖尿病足医学临床研究中心资助项目(2018BCC340);湖北省自然科学基金面上项目(2018CFB756)。

摘　　要：目的:探讨猪脱细胞真皮基质(pADM)和肿瘤坏死因子-α(TNF-α)对毛囊角质细胞Wnt3a蛋白/β-连环蛋白(β-catenin)信号通路表达的影响。方法:①在小鼠背部皮肤用皮肤取样器制作8 mm全层皮肤缺损模型,用带有窗口的8 mm硅胶套环与创面皮缘缝合,分别使用纱布(纱布组)或pADM(pADM组)覆盖创面,观察创面愈合情况,并于第7、14、21天取从硅胶套环窗口处爬行的新生皮肤组织,通过免疫组化法检测皮肤组织中TNF-α、Wnt3a和β-catenin蛋白的表达水平。②在体外实验中,大鼠毛囊角质细胞分为对照组(正常培养)、pADM组、TNF-α组、TNF-α抑制剂组、TNF-α+pADM组、TNF-α抑制剂+pADM组6组,给予相应的干预措施培养48 h,免疫印迹法(Western Blot)检测各组细胞中Wnt3a、β-catenin蛋白表达量。结果:①体内实验结果显示,与纱布组比较,pADM组创面组织中的TNF-α、Wnt3a、β-catenin蛋白表达均上调,分别在第14、7、21天达峰值,各时间点之间差异均有统计学意义(P均<0.01)。②体外实验结果显示,与对照组相比,毛囊角质细胞中的Wnt3a和β-catenin蛋白水平在TNF-α组表达下调,在pADM组、TNF-α抑制剂组、TNF-α+pADM组、TNF-α抑制剂+pADM组表达上调。pADM组与TNF-α抑制剂组比较,Wnt3a和β-catenin蛋白水平差异有显著性(P均<0.01),在pADM组上调水平更高;TNF-α抑制剂组与TNF-α抑制剂+pADM组相比,两蛋白表达水平的差异具有显著性(P均<0.01),TNF-α抑制剂+pADM组上调水平更高。结论:pADM在体内能够上调TNF-α、Wnt3a和β-catenin的蛋白表达,在体外可能具有TNF-α抑制剂样作用,并能加强TNF-α抑制剂的作用。pADM具有促进全皮层缺损创面愈合的作用,并可能通过上调毛囊角质细胞中Wnt3a/β-catenin蛋白的表达促进毛囊再生。Objective:To investigate the effects of porcine acellular dermal matrix(pADM)and tumor necrosis factor-α(TNF-α)on the expression of Wnt3 a protein/β-catenin signaling pathway in hair follicle keratinocytes.Methods:①A 8 mm full-thickness skin defect model was made with punch on the back skin of mice,and a 8 mm silicone loop with a window was sutured to the skin edge of the wound.The wound was covered with gauze(gauze group)and pADM(pADM group)respectively.The wound healing was observed,and the new skin tissue crawling from the window of the silicone collar was taken on the 7,14,and 21 days,and the expression levels of TNF-α,Wnt3 a andβ-catenin in the skin tissue were detected by immunohistochemistry.②In in vitro experiments,rat hair follicle keratinocytes were divided into control group(normal culture),pADM group,TNF-αgroup,TNF-αinhibitor group,TNF-α+pADM group,TNF-αinhibitor+pADM group.All the 6 groups were cultured for 48 hours with corresponding intervention measures,and the Wnt3 a andβ-catenin protein expression levels in the cells of each group were detected by Western blotting.Results:①In vivo experimental results showed that compared with the gauze group,the expressions of TNF-α,Wnt3 a,andβ-catenin protein in the wound tissue of the pADM group were all up-regulated,reaching their peaks on the 14,7 and 21 days,respectively,between each time points the differences were statistically significant(P<0.01).②In vitro experimental results showed that,compared with the control group,the expressions of Wnt3 a andβ-catenin protein in hair follicle keratinocytes were down-regulated in the TNF-αgroup,but were up-regulated in the pADM group,TNF-αinhibitor group,TNF-α+pADM group,and TNF-αinhibitor+pADM group.Compared with the TNF-αinhibitor group,the pADM group showed significant differences in Wnt3 a andβ-catenin protein levels(P<0.01),and the up-regulation level was higher in the pADM group.Compared with the TNF-αinhibitor+pADM group,the TNF-αinhibitor group showed significant differences in

关 键 词：脱细胞真皮基质 肿瘤坏死因子-Α WNT3A β-catenin 毛囊角质细胞 

分 类 号：R-33[医药卫生] R649.9