猪脱细胞真皮基质联合羧甲基纤维素钠对角质形成细胞分泌波形蛋白及锌指蛋白的表达作用  

Effects of porcine acellular dermal matrix combined with sodium carboxymethylcellulose on the ex-pression of vimentin and zinc finger proteins secreted by keratinocytes

作　　者：周兆文 李恭驰[2] 李炳辉[1] 邹利军[1] 李梅 邓海波 Zhou Zhaowen;Li Gongchi;Li Binghui;Zou Li Jun;Li Mei;Deng Haibo(Department of Wound Repair,Liyuan Hospial Affiliated to Tongi Medical College,Huazhong University of Science and Technology,Wuhan 430077,China;Department of Hand Surgery,Union Hospital,Tongi Medical College,Huazhong University of Science and Technology,Wuhan 430022,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,武汉430077 [2]华中科技大学同济医学院附属协和医院手外科,武汉430022

出　　处：《中华实验外科杂志》2024年第9期1988-1991,共4页Chinese Journal of Experimental Surgery

基　　金：2020湖北省重点研发计划项目(2020BCB029);2020湖北省自然科学基金项目(2020CFB696);国家重大疾病多学科合作诊疗能力建设项目国卫办医函[2019]542号。

摘　　要：目的 探讨猪脱细胞真皮基质(pADM)联合羧甲基纤维素钠(CMC)对角质形成细胞上皮-间充质转化(EMT)的影响.方法 体外建立大鼠毛囊角质形成细胞系,实验分组:A组(空白对照)、B 组(2%CMC)、C 组(5%pAD)、D 组(5%pADM+2%CMC).细胞计数试剂盒(CCK-8)检测各组凝胶对角质形成细胞的活力影响.Transwell实验检测角质形成细胞的迁移能力.聚合酶链反应(PCR)和蛋白质印迹法(Western blot)检测各组EMT相关蛋白的表达变化.两组间比较采用非配对t检验,多组间比较采用单因素方差分析.结果 CCK-8实验结果显示,D组角质形成细胞活力最强(A:100.00±8.70、B:124.10±8.10、C:130.40±7.30、D:152.30±5.50),各组间比较差异有统计学意义(F=189.2,P<0.01).Transwell实验中,各组迁移的角质形成细胞数分别为,A组为(95.889±8.638)个、B 组为(149.556±7.091)个、C 组为(172.778±7.710)个、D 组为(236.667±9.695个;与A组比较,各组间比较差异有统计学意义(F=354.9,P<0.01),其中D组角质形成细胞迁移数最多.PCR结果显示,D组角蛋白10(CK10)、波形蛋白(Vim)及锌指蛋白(Snail)表达量最多,分别为4.69、4.25、4.52(F=125.3、146.8、100.3,P<0.05),而 CK14 表达量最低(0.18,F=96.5,P<0.05),差异均有统计学意义.Westem blot结果显示,D组CK10、Vim及Snail蛋白表达量最多,分别为 0.43、0.54、0.69(F=187.5、168.9、151.3,P<0.05),而 CK14 蛋白表达量最低(0.10,F=167.4,P<0.05),差异有统计学意义.结论 pADM及CMC均可改善角质形成细胞生物学活性,pPADM联合CMC可促进CK10、Vim及Snail的表达,并抑制CK14的表达.Objective To investigate the effect of porcine acellular dermal matrix(pADM)com-bined with sodium carboxymethyl cellulose(CMC)on epithelial-mesenchymal transformation(EMT)of ke-ratinocytes and its mechanism.Methods Rat hair follicle keratinocytes were established in vitro and di-vided into four groups:group A(blank control),group B(2%CMC),group C(5%pADM)and group D(5%pADM+2%CMC).The effect of gel on the viability of keratinocytes was detected by cell counting kit(CCK-8)assay.Transwell assay was used to detect the migration ability of keratinocytes.Polymerase chain reaction(PCR)and Western blotting were used to detect the expression of EMT-related proteins in each group.Non-paired t-test was used for comparison between the two groups,and single factor analysis of variance was used for comparison between multiple groups.Results Results The results of CCK-8 assay showed that keratinocyte activity was the strongest in group D(group A:100.0±8.70,group B:124.1±8.10,group C:130.4±7.30,group D:152.3±5.50),and the difference was statistically significant a-mong all groups(F=189.2,P<0.01).In Transwell experiment,the number of migrating keratinocytes was 95.889±8.638,149.556±7.091,172.778±7.710,and 236.667±9.695 in groups A,B,C and D,respectively.There was a statistically significant difference between groups B,C and D and group A(F=354.9,P<0.01).The number of migrating keratinocytes in group D was the greatest.PCR results showed that the expression levels of keratin 10(CK10),Vimentin(Vim)and zinc finger protein(Snail)in group D were the highest(4.69,4.25,4.52;F=125.3,146.8,100.3.all P<0.05)and the expres-sion of CK14 was the lowest(0.18,F=96.5,P<0.05).Western blotting results showed that the expres-sion levels of CK10,Vim and Snail in group D were the highest(0.43,0.54,0.69;F=187.5,168.9,151.3.all P<0.05),while the expression level of CK14 was the lowest(0.10,F=151.3,F=167.4,P<0.05).Conclusion Both pPADM and CMC can improve the biological activity of keratinocytes.pADM combined with CMC can promote the expression o

关 键 词：猪脱细胞真皮基质 羧甲基纤维素钠 角角质形成细胞 上皮-间充质转化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]