Please wait a minute...
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
 全文: PDF(1084 KB)   HTML( 3 )
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
服务  
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章  
李文波
贾丁丁
王飞
张超
石杰
张洪
吴路加
高秋明

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        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
1 TAYLOR G I , PALMER J H . The vascular territories (angiosomes) of the body experimental study and clinical applications[J]. Br J Plast Surg, 1987, 40 (2): 113- 141
doi: 10.1016/0007-1226(87)90185-8
2 MIYAMOTO S , MINABE T , HARⅡ K . Effect of recipient arterial blood inflow on free flap survival area[J]. Plast Reconstr Surg, 2008, 121 (2): 505- 513
doi: 10.1097/01.prs.0000299185.32881.55
3 王天辉, 刘洪涛, 佘晓俊 et al. L-精氨酸对大鼠心肌相对缺血/再灌注损伤保护作用的研究[J]. 中国应用生理学杂志, 2005, 21 (3): 266- 269
WANG Tianhui , LIU Hongtao , SHE Xiaojun et al. Study of the protective effect of L-arginine on relative ischemic/reperfusion injury in myocardium of rat[J]. Chinese Journal of Applied Physiology, 2005, 21 (3): 266- 269
4 杨国斌 . 生物医学研究中实验动物伦理学问题的思考[J]. 中国医学伦理学, 2010, 23 (3): 10- 13
YANG Guobin . Ethical reflection on laboratory animals in biomedical research[J]. Chinese Medieal Ethics, 2010, 23 (3): 10- 13
5 胡斯旺, 庄跃宏, 崔怀瑞 et al. 大鼠背部穿支体区间choke血管变化规律初探[J]. 中国临床解剖学杂志, 2014, 32 (5): 580- 584
HU Siwang , ZHUANG Yuehong , CUI Huairui et al. Study on enlargement law of choke vessels between bilateral iliolumbar perforators in rat[J]. Chinese Journal of Clinical Anatomy, 2014, 32 (5): 580- 584
6 TAYLOR G I , CORLETT R J , DHAR S C et al. The anatomical (angiosome) and clinical territories of cutaneous perforating arteries:development of the concept and designing safe flaps[J]. Plast Reconstr Surg, 2011, 127 (4): 1447- 1459
doi: 10.1097/PRS.0b013e318208d21b
7 CORMACK G C , LAMBERTY B G . A classification of fascio-cutaneous flaps according to their patterns of vascularisation[J]. Br J Plast Surg, 1984, 37 (1): 80- 87
doi: 10.1016/0007-1226(84)90049-3
8 PRASAD V , MORRIS S F . Propeller DICAP flap for a large defect on the back-case report and review of the literature[J]. Microsurgery, 2012, 32 (8): 617- 621
doi: 10.1002/micr.v32.8
9 CHUBB D P , TAYLOR G I , ASHTON M W . True and "Choke" anastomoses between perforator angiosomes:part Ⅱ. Dynamic thermographic identification[J]. Plast Reconstr Surg, 2013, 132 (6): 1457- 1464
doi: 10.1097/01.prs.0000434407.73390.82
10 赵利军, 门秀丽, 董淑云 et al. L-精氨酸对大鼠肢体缺血/再灌注后心肌损伤的影响[J]. 中国药理学通报, 2008, 24 (6): 827- 830
ZHAO Lijun , MEN Xiuli , DONG Shuyun et al. Effect of L-arg on myocardium in rat which suffered from limb ischemia-reperfusion[J]. Chinese Pharmacological Bulletin, 2008, 24 (6): 827- 830
11 GAO Z M , LIN D M , WANG Y et al. Role of the NO/cGMP pathway in postoperative vasodilation in perforator flaps[J]. J Reconstr Microsurg, 2015, 31 (2): 107- 121
doi: 10.1055/s-00000029
12 SILVA J J , POMPEU D G , XIMENES N C et al. Effects of kaurenoic acid and arginine on random skin flap oxidative stress, inflammation, and cytokines in rats[J]. Aesthetic Plast Surg, 2015, 39 (6): 971- 977
doi: 10.1007/s00266-015-0559-8
13 周立纯. L-精氨酸对任意型皮瓣成活的影响的实验研究[D]. 武汉: 武汉大学, 2004.
ZHOU Lichun. Effects of L-arginine on the survival of the random pattern skin flap[D]. Wuhan: Wuhan University, 2004. (in Chinese)
14 LUO Z , SHI H , ZHANG H et al. Plasmid DNA containing multiple CpG motifs triggers a strong immune response to hepatitis B surface antigen when combined with incomplete Freund's adjuvant but not aluminum hydroxide[J]. Mol Med Rep, 2012, 6 (6): 1309- 1314
doi: 10.3892/mmr.2012.1079
15 TAUB P J , SILVER L , WEINBERG H . Plastic surgical perspectives on vascular endothelial growth factor as gene therapy for angiogenesis[J]. Plast Reconstr Surg, 2000, 105 (3): 1034- 1042
doi: 10.1097/00006534-200003000-00031
16 ZHANG F , OSWALD T , LIN S et al. Vascular endothelial growth factor (VEGF) expression and the effect of exogenous VEGF on survival of a random flap in the rat[J]. Br J Plast Surg, 2003, 56 (7): 653- 659
doi: 10.1016/S0007-1226(03)00214-5
17 鲁晓波, 郝建伟, 王兆强 . 早期反复短时缺血训练对皮瓣成活的影响及其机制研究[J]. 中国修复重建外科杂志, 2006, 20 (7): 717- 721
LU Xiaobo , HAO Jianwei , WANG Zhaoqiang . Study on mechanism of the circulation reconstructing of skin flap after early-repeated short ischemia training[J]. Chinese Journal of Reparative and Reconstructive Surgery, 2006, 20 (7): 717- 721
[1] 柳银兰,罗燕,杨文君,施军平,庄振杰. 甘草酸二铵肠溶胶囊对棕榈酸诱导的肝细胞炎症相关基因表达影响[J]. 浙江大学学报(医学版), 2017, 46(2): 192-197.
[2] 白石 等. 雷公藤甲素缓解局灶性脑组织缺血再灌注损伤诱发的炎症及其相关凋亡的研究[J]. 浙江大学学报(医学版), 2016, 45(5): 493-500.
[3] . 七氟醚预处理对大鼠产生延迟相的心肌保护作用[J]. 浙江大学学报(医学版), 2012, 41(5): 553-558.
[4] . 尼克酰胺对帕金森病模型小鼠的保护作用[J]. 浙江大学学报(医学版), 2012, 41(2): 146-152.
[5] . GAD67-GFP基因敲入小鼠GFP与nNOS在嗅球中的表达和共存研究[J]. 浙江大学学报(医学版), 2012, 41(2): 159-165.
[6] 陆建锋,厉旭云,沈春红,陆源,李芝,叶治国,王琼,夏强,王会平. 西红花酸部分内皮依赖性血管舒张作用及其机制[J]. 浙江大学学报(医学版), 2010, 39(6): 559-565.
[7] 徐海潮,钱令波,茹筱晨,缪海锋,叶治国,王会平. 阿托伐他汀对大鼠心肌缺血/复灌电生理的影响[J]. 浙江大学学报(医学版), 2010, 39(6): 589-593.
[8] 傅佳寅,夏满莉,陆建锋,柳琪,蔡新,杨洁,王会平,夏强. 白桦脂酸改善氧化应激损伤大鼠血管的内皮依赖性舒张功能[J]. 浙江大学学报(医学版), 2010, 39(5): 523-529.
[9] 吴海琴,任蓓,沙娟娟,王虎清,张桂莲,李明. 大鼠衰老过程中海马血管内皮生长因子及微血管密度的表达特点[J]. 浙江大学学报(医学版), 2010, 39(4): 404-408.
[10] 李一乔,陈浩浩,尹一飞,韩飞,叶学松,凌树才. 甲醛吸入对大鼠嗅球和海马神经细胞的影响[J]. 浙江大学学报(医学版), 2010, 39(3): 272-277.
[11] 刘晓艳;方红;陈鸿超;蒋筱凌. CMV启动子调控的VEGF-shRNA表达载体的构建及有效干扰序列的筛选[J]. 浙江大学学报(医学版), 2010, 39(2): 181-186.
[12] 李向荣,龙宇红,方晓,刘晓光. 维生素C和E对染铅大鼠海马抗氧化酶及NO与NOS的影响[J]. 浙江大学学报(医学版), 2008, 37(2): 189-192.
[13] 刘伟光;顾伟忠;周银宝;汤宏峰;李民驹. 动脉栓塞化疗对肾母细胞瘤细胞增殖的影响[J]. 浙江大学学报(医学版), 2008, 37(1): 83-87.
[14] 沈毅弘,蔡勇,蓝倩倩,高萱,夏强,虞燕琴. 中枢一氧化氮在躯体传入冲动抑制中枢性升压反应中的作用及机制[J]. 浙江大学学报(医学版), 2007, 36(5): 477-482.
[15] 罗琼,董丽军,黄荷凤. 轻度子宫内膜异位症不孕患者腹腔液和血清NO水平与IVF-ET结局[J]. 浙江大学学报(医学版), 2007, 36(5): 424-428.