Please wait a minute...
浙江大学学报（医学版）
J Zhejiang Univ (Med Sci)  2017, Vol. 46 Issue (6): 656-661    DOI: 10.3785/j.issn.1008-9292.2017.12.13
    
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
Download: HTML( 3 )   PDF(1084KB)
Export: BibTeX | EndNote (RIS)      

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     
Received: 26 October 2017      Published: 25 December 2017
CLC:  R62  
  R-33  
Corresponding Authors: GAO Qiuming     E-mail: dliwenb@163.com;gaoqm001@sohu.com
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
LI Wenbo
JIA Dingding
WANG Fei
ZHANG Chao
SHI Jie
ZHANG Hong
WU Lujia
GAO Qiuming
Cite this article:

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.

URL:

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


外源性L-精氨酸对大鼠背部跨区皮瓣成活的影响

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 
Fig 1 Representing flaps on d7 after operation
Fig 2 Vascular distribution and structure of flaps on d7 after operation
Fig 3 Hematoxylin and eosin staining of Choke Ⅱ microvessels for each group on d7 after operation
Fig 4 Expression of vascular endothelial growth factor (VEGF) in Choke Ⅱ by immunohistochemical staining on d7 after operation
Fig 5 Expression of vascular endothelial growth factor (VEGF) in Choke Ⅱ on d7 after operation by Western blot
[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] LIU Yinlan,LUO Yan,YANG Wenjun,SHI Junping,ZHUANG ZhenJie. Effect of diammonium glycyrrhizinate and phospholipids complex oninflammatory gene expression induced by palmitic acid[J]. J Zhejiang Univ (Med Sci), 2017, 46(2): 192-197.
[2] BAI Shi, SUN Yayi, WU Lijuan, WU Zhongmin, FANG Marong. Tripotolide ameliorates inflammation and apoptosis induced by focal cerebral ischemia/reperfusion in rats[J]. J Zhejiang Univ (Med Sci), 2016, 45(5): 493-500.
[3] . Sevoflurane preconditioning produces delayed cardioprotection effect through up-regulation of inducible nitric oxide synthase in rats[J]. J Zhejiang Univ (Med Sci), 2012, 41(5): 553-558.
[4] . Protective effect of nicotinamide in a mouse Parkinson′s disease model[J]. J Zhejiang Univ (Med Sci), 2012, 41(2): 146-152.
[5] . GAD67-GFP expression and co-localization with bNOS in main olfactory bulb of GAD67-GFP knock-in mouse[J]. J Zhejiang Univ (Med Sci), 2012, 41(2): 159-165.
[6] . Construction of VEGF gene-targeted small interfering RNA mediated by CMV promoter and its silencing effect[J]. J Zhejiang Univ (Med Sci), 2010, 39(2): 181-186.
[7] Yang Zeran, Chen Zengliang. The Influence of Two Tumor Cell Lines on Nitric Oxide Production from Peritoneal Macrophages in Mice[J]. J Zhejiang Univ (Med Sci), 1998, 27(5): 210-213.