A型肉毒毒素对人瘢痕疙瘩成纤维细胞增殖的影响

The effect of botulinum toxin type A on the proliferation of human keloid fibroblasts

目的探讨A型肉毒毒素对人瘢痕疙瘩成纤维细胞增殖的影响。方法在体外分离和培养人瘢痕疙瘩成纤维细胞过程中加入A型肉毒毒素(稀释成50 U/ml)进行干扰。实验组分为2组,各10例,分别加用2种不同剂量的A型肉毒毒素(1×10~6/ml细胞分别加入肉毒毒素1.0μl和2.5μl)并比较其影响效果。空白对照组8例,培养液中不加A型肉毒毒素。共同培养72 h后,采用免疫组化染色(LsAB法)检测细胞Ki67的表达状况。结果实验组:A型肉毒毒素作用后细胞增殖速度较慢,细胞数量变少。空白对照组:细胞Ki67的阳性表达率为63.2%,不同剂量肉毒毒素实验组细胞中Ki67的阳性表达率分别为33.6%和25.6%。空白对照组与实验组组间比较,其差异具有统计学意义(P<0.01);不同剂量A型肉毒毒素实验组组间比较,其差异也具有统计学意义(P<0.05)。结论 A型肉毒毒素可以抑制瘢痕疙瘩组织中成纤维细胞的增殖活性,且药物剂量越大抑制作用可能更明显。

Objective To investigate the effect of botulinum toxin type A （BTXA） on the proliferation of human keloid fibroblasts.Methods Human keloid fibroblasts were isolated and cultured in vitro,and a medium containing BTXA （diluted concentration of 50 U/ml） was used to interfere with cell growth.The experimental group was divided into 2 gToups of 10 cases each.Different amounts of BTXA were added to the cell medium of each group （1.0 μ l and 2.5 μ l）,including 1 × 106 cells.The effect of BTXA on each group was compared.The blank control group （cell medium without BTXA） included 8 cases.At 72 hours after co-culture,immunohistochemistry （LsAB method） was adopted to detect the expression of Ki67.Results The rate of cell proliferation was slow and the number of cells decreased in the experimental group.The positive expression rate of Ki67 was 63.2％ in the blank control group and 33.6％ in the experimental group,showing a significant difference between the two groups （P＜0.01）.The positive expression rate of Ki67 was 33.6％ in the experimental group with the BTXA dose of 1.0 μl and 25.6％ in the experimental group with the BTXA dose of 2.5 μl,showing a significant difference between the two groups （P＜0.05）.Conclusion The proliferation of human keloid fibroblasts can be inhibited by BTXA,and the larger the dosage,the more obvious the inhibition effect.