摘要
目的探讨局部应用外源性热休克蛋白90α(以下简称HSP90α)功能基因"F-5",对大鼠静脉淤血型皮瓣缺血再灌注损伤模型皮瓣成活率的影响。方法通过携带pet15b-F-5重组质粒的构建、大肠杆菌细胞培养扩增,以及HSP90α"F-5"基因表达蛋白的纯化,获得外源性HSP90α;以大鼠腹壁浅血管束为蒂设计轴型皮瓣。将大鼠随机分为3组(每组20只):A组为空白对照组;B组为HSP90α预处理组;C组为HSP90α治疗组。每组又分为2个亚组,根据再灌注时间点分为缺血6 h亚组和缺血8 h亚组。模型建立过程中采用激光多普勒血流仪检测皮瓣血流灌注量,术后每天观察皮瓣的颜色、皮纹、质地、毛发生长、毛细血管充盈试验、针刺出血情况等;比较3组皮瓣的成活率、病理组织学变化以及微血管密度等指标。结果 B组在静脉阻断过程中和再灌注后,皮瓣的血流灌注量较其他2组高;术后7 d,A组皮瓣成活率为(19.3±3.8)%,B组为(85.6±17.6)%,C组为(78.9±20.5)%;B组皮瓣的成活率最高(P<0.05)。术后1、3、7、10、14 d观察,B组及C组皮瓣的炎性细胞浸润较A组明显减少,血管内较少有血栓形成,成纤维细胞及血管内皮细胞增殖显著,新生血管密度增加明显;且缺血6 h亚组的参数均优于8 h亚组。结论建立轴型皮瓣静脉淤血型缺血再灌注损伤大鼠模型,将阻断静脉的时间控制在6 h为宜;外源性HSP90α"F-5"表达蛋白可增加皮瓣血流再灌注量,降低细胞渗透,促进新血管形成,从而对静脉淤血型皮瓣缺血再灌注损伤有较好的治疗作用,而且提前干预的效果更优。
Objective To explore the effect of the local delivery of exogenous heat shock protein 90 α （HSP90 α） functional gene, ＂F-5＂, on the survival ratio of the ischemia-reperfusion injury of venous blood-congested flap in rats. Methods A recombinant plasmid carrying petISb-F-5 was constructed, Escherichia eoli cells were cultured and ampliflcated, the HSP90 α ＂F-5＂ induced protein was purified, and the exogenous HSP90 α was obtained. An axial skin flap pedicled with a superficial vascular bundle of the abdominal wall of rats was designed. The rats were randomly divided into three different intervention groups （20 in each group）： group A was the control group, in which regional flaps were treated with normal saline; group B was the HSP90 α pretreatment group, in which regional flaps were treated with exogenous HSP90α before venous occlusion; and group C was the HSP90 α treatment group, in which regional flaps were treated with HSP90 α immediately after the establishment of an ischemia-reperfusion injury model of venous blood-congested flaps. Additionally, two subgroups were set up with in each group according to the reperfusion time point, namely, ischemia for 6 or 8 hours （10 rats each）. Blood flow perfusion of the skin flap was detected through laser Doppler flowmetry in the process of model establishment. The flaps were observed for color, dermatoglyph, texture, hair growth, acupuncture bleeding, and a capillary filling test was performed. Histological changes including neovascularization and re-epithelialization were compared among the 3 groups. Results In group B, the blood flow perfusion of the skin flap was higher than that of the other 2 groups during the process of venous occlusion and after reperfusion. At 7 days after the operation, the survival rate of the flaps in each group was （19.3 ± 3.8） %, （85.6 ± 17.6） % and （78.9 ± 20.5） %, respectively, in group A, B, and C. The highest survival rate of the flaps was group B （P〈 0.05）. At 1, 3, 7, 10 and 14 days postoperative, inflammatory cell infiltration in group B and C was significantly less than that in group A. Thrombosis was less formed in the blood vessel, and fibroblasts and vascular endothelial cells were significantly proliferated. The neovascularization density increased signiflcandy. Finally, all the parameters studied were better across all the groups when the ischemia time was 6 h instead of 8 h. Conclusion The rat model of isehemia-reperfu-sion injury of venous blood-congested flap was established. The venous occlusion time should be limited to 6 hours. Exogenous HSP90α ＂F-5＂ expression protein can increase flap reperfusion, reducing cell permeability and promoting the formation of new blood vessels.Therefore, it provides better therapeutic efficacy for ischemia-reperfusion injury of venous blood-congested flaps. Early intervention was more effective.
出处
《中国美容整形外科杂志》
CAS
2018年第2期105-110,J0005,共7页
Chinese Journal of Aesthetic and Plastic Surgery
