神经生长因子对大鼠跨区穿支皮瓣成活的影响

doi:10.13418/j.issn.1001-165x.2017.02.009

中国临床解剖学杂志

神经生长因子对大鼠跨区穿支皮瓣成活的影响

杨曦^1，2，　徐永清²，　何晓清²，　王腾²，　李国栋¹

1.第三军医大学研究生管理大队，重庆 400038； 2.成都军区昆明总医院骨科，昆明 650032

收稿日期:2016-09-10 出版日期:2017-03-25 发布日期:2017-04-27
通讯作者: 徐永清，主任医师，教授，博士生导师，E-mail：xuyongqingkm@163.net
作者简介:杨曦（1987-），男，云南曲靖人，在读硕士，研究方向为：显微外科，Tel：（0871）64774526，E-mail：53033239@qq.com
基金资助:
全军后勤科研计划面上项目（CCD14J003）；全军后勤科研计划重大专项（AWS14C003）

Effect of nerve growth factor on survival of cross-boundary perforator flap in rats

YANG Xi ^1,2，Li Guo-dong¹，WANG Teng²，HE Xiao-qing²，XU Yong-qing²

1.Brigade of Postgraduate Management, Third Military Medical University, Chongqing 400038, China; 2. Department of Orthopedics, Kunming General Hospital, PLA, Kunming 650000, China

Received:2016-09-10 Online:2017-03-25 Published:2017-04-27

摘要/Abstract

摘要：

目的　评价神经生长因子（NGF）对SD大鼠背部跨区穿支皮瓣成活的影响。方法　成年SD大鼠60只，平均分为实验组和对照组。切取面积约3cm×10cm的背部跨区皮瓣。实验组皮下注射NGF溶液（10 nmol·ml-1·kg-1）；对照组同法注射0.1 mol/l磷酸盐缓冲溶液（1 ml/kg）。术后第3、7天局部取材运用Western Blot技术半定量检测VEGF及CD34蛋白含量；显微CT微血管三维重建，析因设计分析血管容积及总长度的形态学变化。后术第7天统计皮瓣存活面积，免疫组化染色观测血管生长因子受体(KDR)及NGF受体(TrkA)的表达。结果　第3天，VEGF表达实验组与对照组无显著统计学差异（P=0.088）；实验组CD34表达明显高于对照组(P=0.004)；第7天，两种蛋白分子表达，实验组显著均优于对照组(P<0.05)。血管形态学分析示NGF和时间因相互作用增加血管容积（F=33.304，P<0.05）及总长度（F=8.493，P=0.01）；实验组皮瓣成活面积明显大于对照组（P<0.05）。免疫组化染色示实验组KDR阳性表达较为明显。结论　NGF可促进跨区供血穿支皮瓣的成活。

关键词: 神经生长因子, 跨区皮瓣, 穿支皮瓣, 显微CT, 形态分析

Abstract:

Objective　To evaluate the nerve growth factor (NGF) effect on survival of dorsal cross-boundary perforator flap of SD rats.　Methods　60 adult SD rats were divided into the experimental group and the control group equally，in the dorsum of which a cross-boundary perforator flap with an area of about 3 cm×10 cm was harvested. The experimental group was injected subcutaneously with NGF solution (10 nmol·ml-1·kg-1); the control group was injected with 0.1 mol/L PBS solution (1ml/kg). 3 and 7 days after surgery, VEGF and CD34 of the local tissues of flap were tested via Blot Western；Angiography was performed by the micro-CT to detect Three-dimensional morphological alteration of microvasculature using factorial design analysis of vascular volume and total length. At day 7, survival area of the flap was measured before the observation of expression of KDR and TrkA in vasculature by immunohistochemical stain. Results Day 3, there was no statistically significant differences in the expression of VEGF between two groups (P=0.088); the expression of CD34 in the experimental group was higher than that in the control group (P=0.004); Day 7, there were higher expression of VEGF and CD34 in the experimental group (P<0.05)；Morphological analysis showed that there is an interaction between NGF and time in the increase of vascular volume (F=33.304, P<0.05) and the total length of vessels (F=8.493, P=0.01); Compared with the control group, the flap survival area of the experimental group was significantly higher (P<0.05). The expression of KDR in experimental group was markedly superior to the control group. Conclusion NGF can promote the survival of thecross-boundary perforator flap in SD rat.

Key words: Nerve growth factor, Cross-boundary flap, Perforator flap, Micro-computed tomography, Morphologic analysis

杨曦,徐永清,何晓清,王腾,李国栋. 神经生长因子对大鼠跨区穿支皮瓣成活的影响[J]. 中国临床解剖学杂志, doi: 10.13418/j.issn.1001-165x.2017.02.009.

YANG Xi，Li Guo-dong，WANG Teng，HE Xiao-qing，XU Yong-qing. Effect of nerve growth factor on survival of cross-boundary perforator flap in rats[J]. Chinese Journal Of Clinical Anatomy, doi: 10.13418/j.issn.1001-165x.2017.02.009.

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神经生长因子对大鼠跨区穿支皮瓣成活的影响

Effect of nerve growth factor on survival of cross-boundary perforator flap in rats

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