A型肉毒毒素基于TGF-β1/MAPK通路抑制增生性瘢痕的体外实验研究  

Inhibitory effect of Botulinum toxin type A on hypertrophic scarbased on TGF-β1/MAPK pathway invitro

作　　者：马娟[1] 余扬[1] 于扬[1] 张波[1] 董祥林[1] MA Juan;YU Yang;YU Yang;ZHANG Bo;DONG Xianglin(Department of Plastic Surgery,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院整形科,乌鲁木齐830054

出　　处：《新疆医科大学学报》2023年第5期618-621,共4页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金青年基金项目(2019D01C308)。

摘　　要：目的研究A型肉毒毒素(Botulinum toxin type A,BTXA)对增生性瘢痕成纤维细胞(Hypertrophic scar fibroblasts,HSFbs)的增殖、迁移、凋亡的作用及细胞因子的变化与信号通路的影响。方法切取增生性瘢痕组织,分离培养成增生性成纤维细胞(HSFbs),通过CCK8法检测不同浓度的BTXA对成纤维细胞增殖的影响,划痕试验观察不同浓度的BTXA对成纤维细胞迁移能力的影响,转化生长因子β1(TGFβ1)刺激体外培养的HSFbs,模拟增生性瘢痕的形成机制,用MAPK通路抑制剂或BTXA参与该过程,蛋白印迹法(Western blot)检测B淋巴细胞瘤-2基因(bcl-2)、人型胶原蛋白(col-1)及P-p38MAPK的表达,明确BTXA对MAPK信号通路的影响。结果与空白对照组比较,不同浓度A型肉毒毒素(BTXA)对增生性瘢痕成纤维细胞的增殖抑制率均升高,并随着浓度的升高而增高,差异有统计学意义(药物浓度在2 u/mL时P<0.05,8 u/mL时P<0.01,该浓度细胞增殖抑制率为59%)。不同浓度的BTXA对增生性成纤维细胞的迁移能力均有抑制作用,随着药物浓度升高而抑制作用增强(8 u/mL时,P<0.01)。BTXA对bcl-2、col-1的表达具有降低作用,BTXA降低P-p38的表达,bcl-2、col-1的表达降低与P-p38表达下降、p38MAPK的磷酸化过程受抑制有关。结论BTXA可通过TGFβ1/p38MAPK信号通路抑制HSFbs的形成。Objective The present study aimed to investigate the effects of botulinum toxin type A(BTXA)on the proliferation,migration,and apoptosis of hypertrophic scar fibroblasts(HSFbs),as well as the changes in cytokines and signaling pathways.Methods The hypertrophic scar tissue was cut,and the hypertrophic fibroblasts(HSFbs)were isolated and cultured.The effects of different concentrations of BTXA on the proliferation of fibroblasts were detected by CCK8 method.The effects of different concentrations of BTXA on migration of fibroblasts were observed by scratch test transforming growth factorβ1(TGFβ1)Stimulate HSFbs cultured in vitro to simulate the formation mechanism of hypertrophic scar.MAPK pathway inhibitors or BTXA were used to participate in the process.Western blot was used to detect the expression of BTXA on B-lymphomatoma-2 gene(bcl-2),human collagen(col-1)and the expression of P-p38MAPK,so as to determine the effect of BTXA on MAPK signal pathway.Results Botulinum toxin type A(BTXA)of the different concentrations can inhibit the proliferation of fibroblasts from hypertrophic scars,and the inhibition effect wasincreased with the increasing of drug concentration(8 u/mL,P<0.01,the inhibition rate of cell proliferation at this concentration was 59%).BTXA at the different concentrations inhibited the migration of hypertrophic scar fibroblasts,and the inhibition wasincreased with the increasing of drug concentration(P<0.01 at 8 u/mL).BTXA can reduce the expression of Bcl-2 and Col-1.BTXA can reduce the expression of p-p38.The decreased expression of Bcl-2 and Col-1 was related to the decreased expression of P-p38 and the inhibition of p38MAPK phosphorylation.Conclusion BTXA may pass TGFβ1/p38MAPK signal pathway inhibits the formation of HSFbs.

关 键 词：增生性瘢痕 成纤维细胞 转化生长因子Β1 MAPK信号通路 A型肉毒毒素 

分 类 号：R751[医药卫生—皮肤病学与性病学]