基于TGF-β1/Smad通路探讨A型肉毒毒素对增生性瘢痕的抑制作用及机制  

To Explore the Inhibitory Effect of Type A Botulinum Toxin on Hypertrophic Scar Based on TGF-β_(1)/Smad Pathway

作　　者：张雪[1] 兰东[1] 宁淑华[1] 于思思[1] ZHANG Xue;LAN Dong;NING Shuhua;YU Sisi(Department of Dermatology and Plastic Surgery,Beijing Chao-Yang Hospital,Capital Medical University,Beijing 100043,China)

机构地区：[1]首都医科大学附属北京朝阳医院皮肤与医疗美容科,北京100043

出　　处：《中国美容医学》2022年第5期93-97,共5页Chinese Journal of Aesthetic Medicine

摘　　要：目的:探讨A型肉毒毒素(BTXA)通过TGF-β_(1)/Smad通路对增生性瘢痕成纤维细胞抑制作用及其机制。方法:体外培养增生性瘢痕成纤维细胞,采用0.2、0.4、0.8 U/ml的BTXA处理成纤维细胞,分别作为0.2 U/ml BTXA组、0.4 U/ml BTXA组、0.8 U/ml BTXA组,空白对照组不采用BTXA(0 U/ml)处理,0.8 U/ml BTXA+TGF-β_(1)组用0.8 U/ml BTXA和10 ng/ml的TGF-β_(1)激活剂处理。甲基噻唑基四唑(MTT)法检测细胞活力;流式细胞术检测细胞凋亡率和细胞周期;蛋白质印迹法(Western blot)检测细胞周期蛋白(CyclinD1)、增殖细胞核抗原(PCNA)、Smad2/3、p-Smad2/3蛋白表达。结果:与空白对照组比较,0.2、0.4、0.8 U/mlBTXA显著降低增生性瘢痕成纤维细胞活力和PCNA蛋白表达,差异有统计学意义(P<0.05)。与空白对照组比较,0.2、0.4、0.8 U/ml的BTXA显著提高增生性瘢痕成纤维细胞凋亡率(P<0.05)。与空白对照组比较,0.2、0.4、0.8 U/ml的BTXA显著升高增生性瘢痕成纤维细胞G0/G1期(P<0.05);显著降低细胞S期、G2/M期和Cyclin D1蛋白表达(P<0.05)。与空白对照组比较,0.2、0.4、0.8 U/ml的BTXA显著降低p-Smad2/3蛋白表达(P<0.05)。与0.8 U/ml BTXA组比较,0.8 U/ml BTXA+TGF-β_(1)组显著升高增生性瘢痕成纤维细胞活力、S期、G2/M期和Cyclin D1、PCNA蛋白表达(P<0.05);显著降低细胞凋亡率和细胞G0/G1期(P<0.05)。结论:BTXA通过抑制TGF-β_(1)/Smad通路从而抑制增生性瘢痕成纤维细胞存活,为临床治疗提供了思路。Objective To investigate the inhibitory eff ect of botulinum toxin type A(BTXA)on hypertrophic scar fi broblasts and its mechanism through TGF-β_(1)/Smad pathway.Methods The hypertrophic scar fi broblasts were cultured in vitro,and the fi broblasts were treated with 0.2,0.4,and 0.8 U/ml BTXA,respectively,as the 0.2 U/ml BTXA group,0.4 U/ml BTXA group,and 0.8 U/ml BTXA group.The blank control group was not treated with BTXA(0 U/ml).The 0.8 U/ml BTXA+TGF-β_(1) group was treated with 0.8 U/ml BTXA and 10 ng/ml of TGF-β_(1) activator.Methylthiazolyltetrazole(MTT)method was used to detect cell viability;fl ow cytometry was used to detect cell apoptosis and cell cycle;Western blot was used to detect cell cycle protein(CyclinD1),proliferating cell nuclear antigen(PCNA),Smad2/3.p-Smad2/3 protein expression.Results Compared with the blank control group,0.2,0.4,0.8 U/ml BTXA signifi cantly reduced the viability of hypertrophic scar fi broblasts and the expression of PCNA protein(P<0.05).Compared with the blank control group,0.2,0.4,0.8 U/ml BTXA signifi cantly increased the apoptosis rate of hypertrophic scar fi broblasts(P<0.05).Compared with the blank control group,0.2,0.4,0.8 U/ml BTXA signifi cantly increased the G0/G1 phase of hypertrophic scar fi broblasts(P<0.05);signifi cantly reduced the cell S phase,G2/M phase and Cyclin D1 protein expression(P<0.05).Compared with the blank control group,0.2,0.4,0.8 U/ml BTXA signifi cantly reduced the expression of p-Smad2/3 protein(P<0.05).Compared with 0.8 U/ml BTXA group,0.8 U/ml BTXA+TGF-β_(1) group signifi cantly increased the viability of hypertrophic scar fi broblasts,S phase,G2/M phase and Cyclin D1,PCNA protein expression(P<0.05),signifi cantly reduce the rate of cell apoptosis and cell G0/G1 phase(P<0.05).Conclusion BTXA inhibits the survival of hypertrophic scar fi broblasts by inhibiting the TGF-β_(1)/Smad pathway.

关 键 词：A型肉毒毒素(BTXA) TGF-β_(1)/Smad通路 增生性瘢痕 机制 

分 类 号：R334.5[医药卫生—人体生理学]