新生毛细血管对皮肤成纤维细胞增殖的影响

Effects of new blood capillaries on the proliferation of dermal fibroblasts

目的:研究新生毛细血管对体外培养人皮肤成纤维细胞增殖的影响,探讨新生毛细血管在扩张过程中的作用。方法:体外原代培养正常人皮肤成纤维细胞和人脐静脉内皮细胞,传至第3代用于实验。用50mL/L的胎牛血清培养基培养24h,然后加入不同的培养基,对照组为100mL/L胎牛血清的DMEM培养基,EC组为EC培养基+100mL/L胎牛血清的DMEM培养基,犤3H犦-TDR掺入法反映细胞增殖情况,流式细胞仪检测细胞周期。结果:经内皮细胞培养基孵育48h后,成纤维细胞增殖明显,3H-TDR掺入量EC组犤(758.31±124.09)计数/min犦高于对照组犤(586.20±60.53)计数/min犦(t=4.318,P<0.01);流式细胞仪检测显示,EC组S期细胞比例犤(21.10±7.27)%犦明显高于对照组犤(11.88±3.97)%犦(t=2.728,P<0.05),提示皮肤成纤维细胞分裂、增殖旺盛。结论:皮肤软组织扩张术中,新生毛细血管的形成是引起皮肤成纤维细胞增殖,皮肤面积增加的机制之一。

AIM:To investigate the effects of new blood capillaries on the proliferation of dermal fibroblasts in vitro and the role of new capillaries during the period of dilatation. METHODS:The primary human dermal fibroblasts and human umbilical endothelial cells were cultured in vitro and the third generation was used in the experiment.The fibroblast was cultured for 24 hours in culture medium containing 5 mL/L fetal bovine serum, and then added into different medium:the control group was 100 mL/L DMEM culture medium of fetal bovine serum, EC group was EC culture medium+100 mL/L DMEM culture medium of fetal bovine serum,then TDR method was used to response the proliferation of cell and flow cytometry were applied to observe the cell cycle. RESULTS:Dermal fibroblasts increased significantly when the cells were incubated in the endothelial medium for 48 hours.The volume of TDR in EC group[(758.31±124.09)counting per minute] was higher than that in control group[(586.20±60.53)counting per minute](t=4.318, P< 0.01).The flow cytometer confirmed that cell proportion of S stage in EC group[(21.10±7.27)%]was obviously higher than that in control group[(11.88±3.97)%](t=2.728,P< 0.05)and showed endothelial cell split as well as bloom proliferation. CONCLUSION:The formation of new blood capillary is one of the mechanisms resulting in the proliferation of skin endothelial cells and the increase of skin area during the period of skin parenchyma expanding.