南方医科大学学报

南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (05): 758-.

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不同浓度腐胺对人皮肤成纤维细胞增殖、迁移和凋亡的影响

陈健霞，荣新洲，樊桂成，李松泽，李庆辉   

  • 出版日期:2015-05-20 发布日期:2015-05-20

Effects of different concentrations of putrescine on proliferation, migration and apoptosis
of human skin fibroblasts

  • Online:2015-05-20 Published:2015-05-20

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摘要： 目的探讨不同浓度腐胺对人皮肤成纤维细胞（human skin fibroblasts，HSF）增殖、迁移、凋亡的影响。方法将体外HSF
分为腐胺组和对照组，以含腐胺浓度分别为0.5、1、5、10、50、100、500、1000 μg/ml完全培养基培养细胞设为腐胺组，不添加腐胺
设为对照组。经对照组及不同浓度腐胺组处理的HSF培养24 h后，再以四唑化合物电子耦联显色法（MTS法）测定细胞增殖能
力（用吸光度值表示），Transwell迁移实验测定细胞的迁移能力，流式细胞技术（Flow Cytometry, FCM）Annexin V/PI标记法测
定细胞凋亡率。结果（1）细胞增殖能力（MTS）测定结果显示，0.5、1、5、10 μg/ml 腐胺组HSF的吸光度值较对照组升高（P<
0.01），并且1 μg/ml时达峰值（0.754±0.024）；500、1000 μg/ml腐胺组HSF吸光度值较对照组降低（P<0.01）；50、100 μg/ml腐胺
组HSF的吸光度值与对照组无明显差异（P>0.05）；（2）Transwell迁移实验结果显示，1 μg/ml腐胺组穿过微孔膜的细胞数目较对
照组显著增加（P<0.01），且达到峰值309±21；50 μg/ml及以上浓度腐胺组HSF穿过微孔膜的数目较对照组减少（P<0.05）；0.5、
5、10 μg/ml腐胺组HSF穿过微孔膜的细胞数目与对照组比较无明显差异（P>0.05）；（3）流式细胞凋亡结果显示，0.5、1、5、10 μg/ml
腐胺组HSF凋亡率较对照组显著降低（P<0.05），而且在1 μg/ml时细胞凋亡率达最低值（11.10±0.79）%；100 μg/ml及以上浓度
腐胺组细胞凋亡率较对照组显著升高（P<0.01），而且随着浓度增加凋亡率逐渐升高；50 μg/ml腐胺组与对照组比较细胞凋亡率
无明显差异（P>0.05）。结论微量腐胺可维持细胞正常的迁移能力，显著提高HSF的细胞增殖能力，而较高浓度的腐胺能显著
抑制HSF迁移与增殖，并诱导细胞发生凋亡，提示不同浓度的腐胺会对创面愈合起到完全不同的作用。

Abstract: Objective To explore the effects of different concentrations of putrescine on the proliferation, migration and
apoptosis of human skin fibroblasts (HSF). Methods HSF cultured in the presence of 0.5, 1.0, 5.0, 10, 50, 100, 500, and 1000 μg/
ml putrescine for 24 h were examined for the changes in the cell proliferation, migration, and apoptosis using MTS assay,
Transwell migration assay, and flow cytometry, respectively. Results Compared with the control cells, HSF cultured with 0.5,
1.0, 5.0, and 10 μg/ml putrescine showed significantly increased cell proliferation (P<0.01), and the effect was the most obvious
with 1 μg/ml putrescine, whereas 500 and 1000 μg/ml putrescine significantly reduced the cell proliferation (P<0.01); 50 and
100 μg/ml did not obviously affect the cell proliferation (P>0.05). Putrescine at 1 μg/ml most significantly enhanced the cell
migration (P<0.01), while at higher doses (50, 100, 500, and 1000 μg/ml) putrescine significantly suppressed the cell migration
(P<0.05); 0.5, 5.0, and 10 μg/ml putrescine produced no obvious effects on the cell migration (P>0.05). HSF treated with 0.5, 1.0,
5.0, and 10 μg/ml putrescine obvious lowered the cell apoptosis rate compared with the control group (P<0.01), and the cell
apoptosis rate was the lowest in cells treated with 1 μg/ml putrescine; but at the concentrations of 100, 500, and 1000 μg/ml,
putrescine significantly increased the cell apoptosis rate (P<0.01), while 50 μg/ml putrescine produced no obvious effect on cell
apoptosis (P>0.05). Conclusion Low concentrations of putrescine can obviously enhance the proliferation ability and maintain
normal migration ability of HSF in vitro, but at high concentrations, putrescine can obviously inhibit the cell migration and
proliferation and induce cells apoptosis, suggesting the different roles of different concentrations of putrescine in wound
healing.