传代人皮肤成纤维细胞凋亡与不同压力和作用时间的关系  

Pressure and duration affecting the apoptosis of passaged human skin fibroblasts in vitro

作　　者：李俊华[1,2] 黄忠名[1] 

机构地区：[1]杭州市萧山区中医院骨科七病区,浙江杭州311200 [2]杭州市中医院骨伤科,浙江杭州310007

出　　处：《中华全科医学》2016年第4期543-545,603,共4页Chinese Journal of General Practice

基　　金：浙江省医药卫生科技计划项目(2012KYB169;2013KYB226);浙江省中医药优秀青年人才基金计划(2012ZQ023);浙江省公益性技术应用研究计划(2013c33096);杭州市医学重点专科专病项目计划(20120533Q39);杭州市卫生科技计划(2013B51);杭州市萧山区重点科研项目(2012234)

摘　　要：目的观察不同压力与作用时间下皮肤成纤维细胞的增殖情况,为减少即时皮肤软组织扩张术后皮瓣坏死提供一些理论依据。方法取23岁男性内踝皮肤成纤维细胞培养、传代后,取第6代皮肤成纤维细胞分空白组、40 kPa组(持续40 kPa压力)、60 kPaⅠ组(间断60 kPa压力)、60 kPaⅡ组(持续60 kPa压力)进行压力试验,试验后培养至第7天,检测各组培养液中IL-6含量,使用透射电镜、光镜观察各组内踝皮肤成纤维细胞线粒体、细胞核的变化,使用Image-Pro Plus 6.0图像分析软件对照片中的线粒体、细胞核进行勾勒,计算线粒体、细胞核平均累积光密度(integrated optical density,IOD),并进行比较。结果 60 kPaⅡ组成纤维细胞IL-6的分泌增加,在透镜下可见线粒体减少,40倍光镜下可见细胞增殖受到抑制,与空白组比较差异有统计学意义(P<0.05)。40kPa组与60 kPaⅠ组成纤维细胞,在透镜下可见线粒体受到了损伤,但40倍光镜下可见成纤维细胞的增殖与空白组相近(P>0.05)。结论间断60 kPa压力不会增加皮瓣坏死的几率,更适合在即时皮肤软组织扩张术使用。Objective To observe the effect of different pressure and duration time on the proliferation of skin fibroblast in vitro,provide theoretical basis for decreasing the necrosis of skin flap caused by skin tissue expansion. Methods The fibroblast cells were isolated and cultured in vitro from the skin of medial malleolus of a 23 years old male. The sixth generation fibroblasts were used for pressure test. The experiment included four groups：40 kPa group（ continuous 40 kPa pressure）,60 kPa Ⅰ group（ discontinuous 60 kPa pressure）,60 kPa Ⅱ group（ continuous 60 kPa pressure）,and control group. The level of IL-6 in nutrient fluid was also measured. The shape of mitochondria and nucleus were observed using transmission electron microscope. The average IOD（ integrated optical density） of mitochondria and nucleus of each group were calculated and compared using Plus Image-Pro 6 image analysis software. Results The level of IL-6 of 60 kPa Ⅱgroup increased,the amount of mitochondria decreased and cell proliferation was inhibited with significant differences compared with blank group（ P〈0. 05）. The fibroblast proliferation of 60 kPa Ⅰ group and 40 kPa group were similar with the control group（ P〈0. 05）. Conclusion The risk of skin flap necrosis would not increase under discontinuous 60 kPa pressure. The discontinuous 60 kPa pressure is suitable for the instant tissue expansion surgery.

关 键 词：成纤维细胞 压力 即时扩张皮瓣 手术并发症 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学]