地塞米松及碱性成纤维细胞生长因子联合应用防治超长任意型皮瓣缺血坏死实验探究.doc
《地塞米松及碱性成纤维细胞生长因子联合应用防治超长任意型皮瓣缺血坏死实验探究.doc》由会员分享,可在线阅读,更多相关《地塞米松及碱性成纤维细胞生长因子联合应用防治超长任意型皮瓣缺血坏死实验探究.doc(14页珍藏版)》请在三一文库上搜索。
1、地塞米松及碱性成纤维细胞生长因子联合应用防治超长任意型皮瓣缺血坏死实验探究 摘要目的:探讨联合应用地塞米松和碱性成纤维细胞生长因子(bFGF)对大鼠背部超长任意皮瓣成活的影响,并进一步探讨大鼠血清中TNF-及皮瓣中内皮细胞的细胞间粘附分子1(ICAM-1)发挥作用的机制。方法:采用大鼠背部皮瓣模型,术后即刻腹腔内一次性大剂量注射地塞米松,皮瓣下均匀滴注bFGF,术后24h内观察TNF-及ICAM-1的变化,术后7天观察皮瓣成活率及组织学变化。结果:皮瓣成活率联合用药组(58.25.2)%bFGF组(50.53.9)%地塞米松组(48.74.2)%对照组(39.53.7)%,(F=32.08,P
2、0.01);联合用药组与地塞米松组较对照组及bFGF组血清中TNF-含量达高峰时间更晚,峰值更低,且各组均于术后24h下降明显;各组于术后4h皮瓣组织的上皮细胞、血管内皮细胞和成纤维细胞的胞浆中均呈现ICAM-1的mRNA表达,并均于术后8h呈现ICAM-1阳性蛋白表达高峰, 对照组及bFGF组明显高于联合用药组和地塞米松组(P0.01),之后缓慢下降,术后12h仍保持较高水平。结论:地塞米松和bFGF均能促进皮瓣成活,且地塞米松+bFGF促进皮瓣成活优于单用地塞米松和单用bFGF。地塞米松能够延迟血清中TNF-达到峰值的时间,显著降低其峰值含量,并通过抑制皮瓣中ICAM-1,干预炎症反应起始
3、过程,从而减轻皮瓣炎症反应。 关键词地塞米松;碱性成纤维细胞生长因子;皮瓣;缺血坏死;成活率;TNF-;ICAM-1 中图分类号R622文献标识码A文章编号1008-6455(2010)04-0591-04 Experimental study of the application that Dexamethasone combined with basic fibroblast growth factor(bFGF) to prevent ischemia and necrosis from ultra-long radom skin flap LIN Jie,QIU Shu-lin (De
4、partment of Plastic Surgery,Hebei PeopleS Hospital,Shijiazhuang 050051,Hebei,China) Abstract:ObjectiveTo study the combined effects of Dexamethasone and basic fibroblast growth factor (bFGF) of the random flap survival on rats dorsal, and further explore the levels of TNF- in serum and how is the me
5、chanism that the intercellular adhesion molecule-1 (ICAM-1) in endothelial cells of skin flap to play a role.MethodsThe rat dorsal skin flap model, a one-time immediately after high-dose intraperitoneal injection of Dexamethasone, drip bFGF equably under the flap, observed the changes of TNF- and IC
6、AM-1within 24h, post 7 days view the survival rate of the flap and histological changes. Results The survival rate of flap combined group (58.2 5.2)% bFGF group (50.5 3.9)% Dexamethasone group (48.7 4.2)% the control group (39.5 3.7)%, (F = 32.08, P0.01). The levels of TNF- in serum of combined grou
7、p and Dexamethasone group reached a peak time later than in the control group and bFGF group, and the peak is lower, and all groups were significantly decreased after 24h; each group 4 hours after surgery, epithelial cells of flap, endothelial cells and fibroblasts were present in the expression of
8、mRNA of ICAM-1 in cytoplasm, and the positive expression peak of ICAM-1were presented at 8h after. In the control group and bFGF group was significantly higher than in the combination group and the Dexamethasone group (P0.01), followed by a slow decline, the level remained high after 12h. Conclusion
9、 The Dexamethasone and bFGF all can promote the flaps survival rate, and Dexamethasone + bFGF group is superior than Dexamethasone alone or bFGF alone. Dexamethasone could delayed the peak time of TNF- in serum, significantly reduced the peak levels, and by inhibiting the ICAM-1 in the flap, interfe
10、re with initiation of inflammation, thus reducing the flap inflammation. Key words: Dexamethasone; basic fibroblast growth factor(bFGF); flap; ischemia and necrosis; survival rate;TNF-;ICAM-1 皮瓣缺血坏死是一种复杂的病理生理过程。伴随随意型皮瓣形成,皮瓣边缘及基底部血供完全离断,仅保留蒂部的血液供应,直接导致血管损伤,血小板聚集,皮瓣组织缺血、缺氧,同时发生白细胞粘附、游走,炎性因子释放,氧自由基损害和钙
11、离子超载等病理变化,而炎症的发生又进一步加重了皮瓣组织损伤。超长随意型皮瓣由于超过了随意型皮瓣长宽比例的最高限度(2:1),蒂部血液供应不能达到皮瓣远段,使得皮瓣远段组织长时间的缺血缺氧情况未得到改善,故其最终的结果是皮瓣远段发生坏死。因此,皮瓣坏死防治的根本措施在于改善血液循环,减轻炎症反应和防止或减轻缺血再灌注损伤。本研究试图通过观察应用地塞米松及碱性成纤维细胞生长因子(bFGF)后大鼠皮瓣成活率、组织学等的变化,同时测定血清中TNF-含量、皮瓣中ICAM-1含量,进一步了解联合应用地塞米松及bFGF提高皮瓣成活率的可行性,以扩大任意皮瓣移植的应用范围。 1材料和方法 1.1 动物来源及分
12、组:健康Wistar大鼠(河北医科大学动物实验中心提供,体重280g10g)96只,雌雄不限。大鼠背部超长任意皮瓣模型制备后,按术后即刻给药不同,将大鼠随机分为四组,即A组为对照组:腹腔注射生理盐水0.2ml,皮瓣下滴注生理盐水0.25ml;B组为地塞米松组:腹腔注射地塞米松0.2ml(用量为5mg/kg),皮瓣下滴注生理盐水0.25ml;C组为bFGF组:腹腔注射生理盐水0.2ml,皮瓣下滴注含1750IU bFGF的生理盐水0.25ml(用量为155IU/cm2);D组为地塞米松联合bFGF组:腹腔注射地塞米松0.2ml,皮瓣下滴注含1750IU bFGF的生理盐水0.25ml。 1.2
13、大鼠背部超长任意皮瓣模型制备:96只大鼠均以2%戊巴比妥钠腹腔注射(3540mg/kg)麻醉,麻醉满意后,背部脱毛,俯卧位固定,碘酒、酒精消毒。于鼠背正中以髂嵴连线为基准线,以后正中线为轴线,掀起一7.5cm1.5cm大小的任意皮瓣(含脂膜肌层,长宽比5:1),其蒂在尾部。3-0丝线原位缝合皮瓣,B、D组术后即刻一次性腹腔注射地塞米松,C、D组术后即刻于皮瓣下均匀滴注 bFGF。A组给予生理盐水腹腔注射及皮瓣下滴注,时间方法同上。 1.3 观测指标及方法:皮瓣大体观察:术后第7天对大鼠背部随意皮瓣成活的情况进行肉眼观察;皮瓣成活率:坏死皮瓣以其外观、颜色及质地来判断,术后7天,用数码相机进行皮
14、瓣摄像,图像输入计算机,应用Image-Pro Plus.v6.0图像分析系统,计算出皮瓣成活率(即皮瓣成活面积与总面积之比);组织学检查:术后3天和7天各组均于皮瓣中段取 0.5cm0.2cm全厚皮瓣组织,4%多聚甲醛固定,石蜡包埋切片、HE染色;a) 光镜下观察组织结构改变;b)进行毛细血管计数,血管计数方法:所有血管均计数在内,每组5张切片,每张切片在中央区及周边区随机选取5个高倍视野,血管计数后求其平均数;c) 进行成纤维细胞计数:每组5张切片,每张切片在中央区及周边区随机选取5个高倍视野,成纤维细胞计数后求其平均数; 检测血清中TNF-含量:各组分别于术后0h、1h、4h、8h、12
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- 地塞米松 碱性 纤维 细胞 生长因子 联合 应用 防治 超长 任意 型皮瓣 缺血 坏死 实验 探究
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