摘要
目的研究脂多糖(LPS)对正常皮肤成纤维细胞的影响及其在瘢痕形成中的作用。方法选取2017年11月-2018年11月在西京医院烧伤与皮肤外科手术切除的增生性瘢痕组织及余留的正常皮肤组织,在建立瘢痕形成细胞模型及小鼠创面愈合模型的基础上,应用HE染色、Masson染色、免疫组化、反转录实时定量PCR(RT-qPCR)、Western blotting、电镜技术观察增生性瘢痕组织及其成纤维细胞特征,分析LPS作用下正常皮肤成纤维细胞纤维化相关因子Ⅰ型胶原(ColⅠ)、Ⅲ型胶原(ColⅢ)及α-平滑肌肌动蛋白(α-SMA)表达的变化,观察细胞亚显微结构、愈合创面外观及真皮结构的改变。结果增生性瘢痕组织中成纤维细胞数目及胞外基质蛋白水平明显高于正常皮肤组织(P<0.05);增生性瘢痕组织及其成纤维细胞纤维化相关因子的表达在mRNA水平及蛋白水平均较正常皮肤组织及其成纤维细胞增高,差异有统计学意义(P<0.05);LPS能够明显提高正常皮肤成纤维细胞ColⅠ、ColⅢ及α-SMA的表达水平(P<0.05),引起内质网亚显微结构的改变,并延缓小鼠创面的愈合,增加愈合创面的瘢痕化。结论LPS可使正常皮肤成纤维细胞内质网发生改变,胞外基质蛋白过度合成及沉积,引起正常皮肤成纤维细胞向增生性瘢痕成纤维细胞表型转化,导致增生性瘢痕的形成。
Objective To study the effect of lipopolysaccharide(LPS)on normal skin tissues(NS)-derived fibroblasts(NSFs)and the formation of hypertrophic scar(HS).Methods HSs and the corresponding NS tissues were sampled from the patients who underwent surgical excision at Xijing Hospital from November 2017 to November 2018.The characteristics of HSs and HSFs were identified by HE and Masson staining,immunohistochemistry/immunocytochemistry,real time quantitative PCR(RTqPCR),Western blotting and electron microscopy in scar formation cell models and mouse wound healing models.Results The staining results showed that the number of fibroblasts,level of extracullar matrix(ECM)proteins(ColⅠ,ColⅢ,andα-SMA)in HS were significantly higher than those in NS(P<0.05).The mRNA and protein expression levels of ColⅠ,ColⅢandα-SMA in HS/HSFs were significantly higher than those in NS/NSFs(P<0.05).LPS could significantly increase the expression of ColⅠ,ColⅢandα-SMA in NSFs,and the result was similar to those in HSFs(P<0.05).The endoplasmic reticulum was normal and regulated in NSFs,but it was swollen and unregulated in LPS-stimulated NSFs and HSFs.After injection of LPS,the mice showed delayed wound healing and obvious scarring.Conclusions LPS can change the ultrastructure of endoplasmic reticulum,increase the expression of fibrotic proteins(e.g.ColⅠ,ColⅢandα-SMA),lead to the accumulation of ECM proteins and the transformation of fibroblasts,and finally form HS.
作者
石珊
李梦凡
王玲
古兰
田雨欣
舒欢
Shi Shan;Li Meng-Fan;Wang Ling;Gu Lan;Tian Yu-Xin;Shu Huan(Department of Burns and Cutaneous Surgery,Xijing Hospital,Air Force Medical University,Xi’an 710032,China)
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2020年第6期604-610,共7页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金(81571914)。
关键词
脂多糖
增生性瘢痕
成纤维细胞
转化
创面愈合
lipopolysaccharide
hypertrophic scar
fibroblast
transformation
wound healing
作者简介
石珊,医学学士,主管护师,主要从事烧伤整形研究及护理工作。E-mail:458680501@qq.com;通信作者:李梦凡,E-mail:mengfan898@qq.com。
