高压氧预处理对大鼠腹部皮瓣ICAM-1、VCAM-1、NF-κB表达及成活的影响

The effect of hyperbaric oxygen preconditioning on the expression of ICAM-1, VCAM-1, NF-κB and flap survival rate during ischemia-reperfusion injury in rat abdominal skin flap

目的探讨高压氧预处理对大鼠腹部皮瓣缺血再灌注损伤模型中胞间黏附分子1（ICAM-1）、血管细胞黏附分子1（VCAM-1）、NF-κB的表达及皮瓣存活的影响。方法采用随机数表法将18只成年雄性SD大鼠分为假手术组（SH）6只、缺血再灌注组（IR）6只和高压氧预处理组（HBO）6只，HBO组术前3d开始行高压氧治疗，每天2次，间隔12h，而IR组和SH组大鼠术前不做处理，3组大鼠均于腹部形成6cm×9cm右侧腹壁浅动脉为蒂的轴型皮瓣，SH组大鼠直接行皮瓣原位缝合，IR组和HBO组加行缺血处理3h。术后第3天取存活皮瓣组织进行HE染色、免疫组化染色，以及Western Blot检测其ICAM-1、VCAM-1和NF-κB的表达。数据采用单因素方差分析，应用SPSS18．0进行统计学分析。结果IR组大鼠腹部皮瓣存活面积百分比为（22．38±4．35）％，平均血流量为（32．61±5．68）PU，HBO组皮瓣存活面积为（45．34±3．15）％，平均血流量为（61．78±3．61）PU，2组比较差异均有统计学意义（P〈0．01）。HE染色可见，SH组炎性细胞浸润程度极低，而IR组炎性细胞浸润程度明显高于HBO组。免疫组化结果中可见，IR组皮瓣ICAM-1、VCAM-1及NF-κB表达呈强阳性，且阳性细胞数明显增多，分别为（47．89±7．48）％、（75．71±5．48）％和（78．17±4．37）％，而HBO组呈弱阳性，阳性细胞数显著减少，分别为（18．59±2．63）％、（28．44±4．69）％和（43．73±9．01）％，2组比较差异有统计学意义（P〈0．01）。Western Blot结果与免疫组化结果基本-致，在HBO组中ICAM-1、VCAM-1及NF-κB的表达量显著下降，而IR组3种因子的蛋白表达量较HBO组明显升高（P〈0．01）。结论高压氧预处理可以降低大鼠腹部皮瓣缺血再灌注损伤过程中ICAM-1、VCAM-1以及NF-KB的表达，并促进皮瓣存活。

Objective To evaluate the effect of hyperbaric oxygen preconditioning on the expression of intercellular adhesion molecule-1 （ ICAM-1 ） , vascular cell adhesion molecule-1 （ VCAM-1 ） , NF-κB and flap survival rate during Isehemia-Reperfusion Injury in Rat Abdominal Skin Flap. Methods 18 male adult SD rats were divided randomly into sham group （SH） , ischemia-reperfusion group （IR） and hyperbaric oxygen preconditioning group （HBO） , 6 rats in each group. The rats in HBO group received regular hyperbaric oxygen treatment for three days （twice a day with an interval of 12 h） before operation. An abdominal skin flap pedicled with superficial epigastric artery was established. Ischemia lasted for 3 hours in the IR group and HBO group. On the third postoperative day, samples were taken to evaluate the expression of ICAM-1 , VCAM-1 and NF-κB by immunohistochemistry staining anti Western Blot. Results In the IR group, the flap survival rate and average blood perfusion were （ 22.38 ±4.35 ） % and （ 32.61 ±5.68） PU, while in the HBO group they were （45.34 ±3. 15） % and （61.78 ±3.61） PU, showing signifieant difference between the two groups （ P 〈 0.01 ）. The inflammatory cell infiltration condition in IR group was much higher than that in the HBO group from the HE staining observation. Compared with the HBO group, ICAM-1 , VCAM-1 anti NF-κB showed （ ＋ ＋ ＋ ） with higher positive cells in the IR group （P 〈 0. 01 ）. Results of Western Blot showed similar conclusion. The relative expression of ICAM-1 , VCAM-1 and NF-κB was much higher in the IR group than that in the HBO group （ P 〈 0.01 ）. Conclusions Hyperbaric oxygen preconditioning could decrease the expression of ICAM-1 , VCAM-1 , NF- κB and promote flap survival rate during the process of ischemia-reperfusion injury on a rat abdominal skin flap model.