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浙江大学学报（医学版）
浙江大学学报（医学版）  2017, Vol. 46 Issue (6): 656-661    DOI: 10.3785/j.issn.1008-9292.2017.12.13
原著     
外源性L-精氨酸对大鼠背部跨区皮瓣成活的影响
李文波1,2(),贾丁丁1,2,王飞1,张超1,2,石杰1,张洪1,2,吴路加3,高秋明1,*()
1. 兰州军区兰州总医院全军骨科中心创伤骨科, 甘肃 兰州 730050
2. 甘肃中医药大学研究生院, 甘肃 兰州 730000
3. 南方医科大学研究生院, 广东 广州 510515
Effect of exogenous L-arginie on survival of extended dorsal perforator flaps in rats
LI Wenbo1,2(),JIA Dingding1,2,WANG Fei1,ZHANG Chao1,2,SHI Jie1,ZHANG Hong1,2,WU Lujia3,GAO Qiuming1,*()
1. Orthopedic Center, Lanzhou General Hospital of PLA, Lanzhou 730050, China
2. Graduate School, Gansu University of Traditional Chinese Medicine, Lanzhou 730000, China
3. Graduate School, Southern Medical University, Guangzhou 510515, China
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摘要：

目的: 观察外源性L-精氨酸对大鼠背部跨区皮瓣成活的影响。方法: 雄性SD大鼠16只随机分为L-精氨酸组和对照组，每组8只，分别建立背部三穿支体跨区皮瓣。L-精氨酸组分别于术前1 d、术后即刻、术后1~7 d腹腔注射L-精氨酸400 mg·kg-1·d-1，对照组于相同时间点腹腔注射等体积等渗氯化钠溶液。术后7 d观察血管走形和分布，计算皮瓣成活率；HE染色观察Choke Ⅱ区新生血管并计算新生血管数量和管径；免疫组织化学染色和蛋白质印迹法检测Choke Ⅱ区血管内皮生长因子（VEGF）表达。结果: 术后7 d，L-精氨酸组有1只大鼠皮瓣全部成活，其余7只在Choke Ⅱ区远端出现小面积不同程度坏死；对照组大鼠皮瓣均有不同程度坏死，坏死范围包括邻近Choke Ⅱ区和全部Choke Ⅱ区以远的皮瓣。术后7 d，两组ChokeⅠ区血管均达到真性吻合，L-精氨酸组Choke Ⅱ区新生血管较多，皮瓣末端血管结构较完整，对照组Choke Ⅱ区新生血管较少，皮瓣末端发黑坏死，观察不到血管结构。L-精氨酸组皮瓣成活率为（88.42±4.19）%，显著高于对照组（76.52±5.37）%（t=3.707，P < 0.01）。术后7 d，L-精氨酸组Choke Ⅱ区新生血管数量和管径分别为（29.47±5.28）个/mm2和（47.27±5.32）μm，明显高于对照组（t=2.694和2.389，P < 0.05或P < 0.01）。免疫组织化学染色和蛋白质印迹法检测结果显示，L-精氨酸组Choke Ⅱ区VEGF表达量明显高于对照组（t=9.428和-3.054，P < 0.05或P < 0.01）。结论: 外源性L-精氨酸可促进大鼠背部跨区皮瓣Choke Ⅱ区血管新生和扩张，改善皮瓣血供，提高皮瓣成活率。
关键词： 硝基精氨酸/药理学;  外科皮瓣/血液供给;  新生血管化, 生理性/药物作用;  血管内皮生长因子A;  一氧化氮;  大鼠, Sprague-Dawley    
Abstract:

Objective: To investigate the effect of exogenous L-Arg on the survival of extended perforator flap in rats. Methods: Sixteen male Sprague Dawley rats were randomly divided into L-Arg group (n=8) and control group(n=8). The extended dorsal three-vascular territory perforator flaps were made in rats. L-Arg (400 mg·kg-1·d-1) was injected intraperitoneally in L-Arg group 1d before operation, immediately and 1-7 d after operation, while the same volume of saline was injected intraperitoneally in control group at the same time points. The appearance and distribution of blood vessels were observed, and the flap survival areas were measured 7d after operation. The tissue samples were harvested from choke zone Ⅱ for histological study and the expression of vascular endothelial growth factor (VEGF) was detected by immunohistochemistry and Western blot, respectively. Results: After 7d, the clearer vascular structure and more new vessels in choke zone Ⅱ were observed in L-Arg group. The survival rate of flap in L-Arg group was (88.42±4.19)%, which was significantly higher than that in control group[(76.52±5.37)%, t=3.707, P < 0.01]. The microvessel density and caliber of choke zone Ⅱ in L-Arg group was (29.47±5.28)/mm2 and(47.27±5.32)μm, which were significantly higher than those in control group (t=2.694 and 2.389, P < 0.05 or P < 0.01). The immunohistochemistry and Western blot showed that the expression of VEGF in choke zone Ⅱ of L-Arg group was significantly higher than that in control group (t=9.428 and -3.054, P < 0.05 or P < 0.01). Conclusion: Exogenous L-Arg can increase the survival rate of extended dorsal perforator skin flap through promoting vascularization and dilatation of vessels in choke zone Ⅱ in rats.
Key words: Nitroarginine/pharmacology    Surgical flaps/blood supply    Neovascularization, physiologic/drug effects    Vascular endothelial growth factor A    Nitric oxide    Rats, sprague-dawley
收稿日期: 2017-10-26 出版日期: 2017-12-25
CLC:  R62  
基金资助: 甘肃省自然科学基金(1506RJZA298);甘肃省卫生行业科研计划(GSWSKY2017-30)
通讯作者: 高秋明     E-mail: dliwenb@163.com;gaoqm001@sohu.com
作者简介: 李文波(1992-), 男, 硕士研究生, 主要从事创伤骨病和骨髓炎诊治研究; E-mail:dliwenb@163.com; https://orcid.org/0000-0002-1812-9457
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引用本文:

李文波,贾丁丁,王飞,张超,石杰,张洪,吴路加,高秋明. 外源性L-精氨酸对大鼠背部跨区皮瓣成活的影响[J]. 浙江大学学报（医学版）, 2017, 46(6): 656-661.

LI Wenbo,JIA Dingding,WANG Fei,ZHANG Chao,SHI Jie,ZHANG Hong,WU Lujia,GAO Qiuming. Effect of exogenous L-arginie on survival of extended dorsal perforator flaps in rats. J Zhejiang Univ (Med Sci), 2017, 46(6): 656-661.

链接本文:

http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2017.12.13    或    http://www.zjujournals.com/med/CN/Y2017/V46/I6/656

图 1  术后7 d两组大鼠皮瓣成活情况
图 2  术后7 d两组大鼠皮瓣血管走形和分布
图 3  术后7 d两组Choke Ⅱ区血管新生情况(HE染色)
图 4  术后7 d免疫组织化学染色观察两组Choke Ⅱ区血管内皮生长因子(VEGF)表达程度
图 5  术后7 d蛋白印迹法检测两组皮瓣Choke Ⅱ区血管内皮生长因子(VEGF)表达程度
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