秋水仙碱对人瘢痕疙瘩成纤维细胞增殖及细胞周期的影响

Effect of colchicine on human keloid fibroblasts

摘要:

目的：研究秋水仙碱对体外培养人瘢痕疙瘩成纤维细胞（keloid Fibroblast，KFB）的毒性作用及细胞周期的影响。方法：体外培养人KFB，不同浓度秋水仙碱（1、2、4、8、16、32 μg/ml）处理细胞。采用MTT比色法检测KFB的毒性作用；采用流式细胞仪检测秋水仙碱对KFB的细胞周期（G2期）的影响。结果：MTT法结果显示1 μg/ml组（12.56±0.79）%，2 μg/ml组（27.35±0.50）%，4 μg/ml组（34.36±0.25）%，8 μg/ml组（39.38±0.57）%，16 μg/ml组（40.38±0.23）%，32 μg/ml组（44.65±0.60）%（F=2440.178，P=0.000）；流式细胞仪检测结果显示空白对照组（0.67±0.46）%，1 μg/ml组（4.26±1.51）%，2 μg/ml组（24.51±4.69）%，4 μg/ml组（45.12±2.75）%，8 μg/ml组（55.81±5.04）%，16 μg/ml组（69.90±3.61）%，32 μg/ml组（85.53±2.06）%，F=491.609，P=0.000。秋水仙碱能抑制细胞的生长，对细胞分裂前期（G2期）产生影响，并呈时间依赖性和药物浓度依赖性。结论：在24 h时间点，秋水仙碱在1～32 μg/ml浓度范围内可调控细胞的生长及细胞周期，可以抑制瘢痕疙瘩的形成。

Abstract:

Objective：To investigate the effect of colchicine on proliferation and cell cycle（G2）of human KFB. Methods：The primary human keloid-derived fibroblast were cultured by tissue. The proliferative behavior of KFB was studied after pre-treating with various concentrations of colchicines（1，2，4，8，16，32 μg/ml） by 4 methyl thiazolyl tetrazolium method and the cell cycle was assessed by flow cytometry. Results：MTT analysis showed that 1 μg/ml group（12.56±0.79）%，2 μg/ml group（27.35±0.50）%，4 μg/ml group（34.36±0.25）%，8 μg/ml group（39.38±0.57）%，16 μg/ml group（40.38±0.23）%，32 μg/ml group（44.65±0.60）%，F=2440.178，P=0.000；flow cytometry analysis showed that blank control group（0.67±0.46）%，1 μg/ml group（4.26±1.51）%，2 μg/ml group（24.51±4.69）%，4 μg/ml group（45.12±2.75）%，8 μg/ml group（55.81±5.04）%，16 μg/ml group（69.90±3.61）%，32 μg/ml group（85.53±2.06）%，F=491.609，P=0.000. Not only was the proliferation of KFB was inhibited，but also the prophase of cell division（G2） was af－fected depending on the time and drug concentration. Conclusion：The primary human keloid-derived fibroblast should be cultured by tissue adherent culture method and colchicine may regulate the proliferation and prophase of cell division of KFB at 1-32 μg/ml in 24 hour.