A型肉毒素预防人瘢痕成纤维细胞增生机制的初步研究

目的 探究不同浓度A型肉毒素(BTX-A)对瘢痕成纤维细胞的影响,初步阐释BTX-A治疗瘢痕及预防术后瘢痕增生的相关分子作用机制.方法 选取人瘢痕成纤维细胞,以不同浓度的B T X-A(0.01、0.10、1.00 U/L及10.00 U/L)作用24 h后,激光共聚焦显微镜观察细胞黏附及骨架变化,并采用MTT及流式技术检测其增殖、凋亡及周期变化,同时采用实时荧光定量PCR(qRCR)及蛋白免疫印迹(Western blot)方法 ,探究TGF-β、基质金属蛋白酶(MMP)-1、MMP-2及MMP-9基因及蛋白的表达变化.结果 随着BTX-A剂量的升高,其细胞黏附数量及骨架荧光强度逐渐减弱;细胞增殖能力减弱且主要阻断在细胞G0/G1期;此外其凋亡也随着BTX-A剂量增大而逐渐增强.qPCR及Western blot结果显示,随着BTX-A剂量增大,MMP-1及MMP-2基因及蛋白均呈现高表达,而TGF-β及MMP-9呈现出低表达.结论 BTX-A通过阻断瘢痕细胞G0/G1期抑制其增殖,同时提高MMP-1及MMP-2的表达来减轻瘢痕形成,对瘢痕的治疗起着积极的作用.

A preliminary study on the mechanism of botulinum toxin type A in preventing the proliferation of keloid fibroblast cells

Objective To investigate the effects of different concentrations of botulinum toxin type A on hypertrophic scar fi-broblasts ,and to explore the molecular mechanism of botulinum toxin type A in the treatment of scar and prevention of postopera-tive scar hyperplasia .Methods Different concentrations of botulinum toxin A (0 .01 ,0 .1 ,1 U/L and 10 U/L) were used on hyper-trophic scar fibroblasts for 24 hours ,to observe the changes of cell adhesion and cytoskeleton under laser confocal microscopy .MTT and flow cytometry were used to detect the proliferation ,apoptosis and cycle of change ,at the same time real time fluorescence quantitative PCR and Western blot were conducted to detected the expression of TGF-β,matrix metalloproteinase MMP-1 ,MMP-2 and MMP-9 gene and protein expression changes .Results With the increase of botulinum toxin A dose ,the number of cell adhesion and cytoskeletal fluorescence intensity decreased ,cell proliferation ability decreased and mainly blocked at G0-G1 phase ,and the ap-optosis also increased with the dose increased .The results of qPCR and Western blot showed that MMP-1 and MMP-9 gene and protein were highly expressed with the increase of botulinum toxin A dose ,while TGF-βand MMP-9 showed low expression .Con-clusion Botulinum toxin A can inhibit the proliferation of hypertrophic scar fibroblasts and inhibit the expression of MMP-1 and MMP-2 ,which can inhibit scar formation .It plays a positive role in the treatment of scar .