EFFECT OF NATURAL HIRUDIN ON SURVIVAL OF DORSAL RANDOM FLAP AFTER EARLY PEDICLE DIVISION IN RATS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

HAN Zhiqiang , YIN Guoqian , WEI Shuyi , LIN Bojie , PAN Xinyuan.

Department of Plastic and Aesthetic Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning Guangxi, 530021, P.R.China. Corresponding author: YIN Guoqian, E-mail: yingq61@163.com;

Corresponding author：

Keywords：

Natural hirudin; Random flap; Early pedicle division; Rat

DOI：

10.7507/1002-1892.20130164

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Objective To explore the effects of natural hirudin on the survival of dorsal random flap after early pedicle division in rats. Methods Thirty-eight adult Wistar rats, male or female, weighing 220-280 g, were selected and randomly divided into natural hirudin group (experimental group, n=19) and normal saline group (control group, n=19). The dorsal random flap of 9 cm × 3 cm in size was prepared on the back of the rats. Six symmetrical injection points were selected at 2, 4, and 8 cm from the pedicle, 0.5 mL (3 ATU) natural hirudin and equivalent normal saline were injected in experimental group and control group respectively every day to the end of the experiment. After 4 days, the pedicles were cut. The flap was observed, and the ratio of flap survival area was measured at 7 days after pedicle division. At immediately after flap preparation, at 1 day before pedicle division, and at 3 and 7 days after pedicle division, the specimens were harvested for histological staining, immunohistochemical staining, microvessel density (MVD) measurement, and detection of vascular endothelial growth factor (VEGF) mRNA expression by fluorescent quantitative RT-PCR. Results All rats of 2 groups survived to the completion of the experiment. After pedicle division, the flap necrosis area of experimental group was smaller than that of control group; at 7 days, the ratio of flap survival area in experimental group was 85.366% ± 2.872%, and was significantly higher than that in control group (75.252% ± 3.455%) (t=7.117, P=0.000). Histological staining showed that no significant difference was found in the number of capillaries between 2 groups at immediately after flap preparation; at 1 day before pedicle division and at 3 and 7 days after pedicle division, the capillary number of experimental group was significantly more than that of control group. Immunohistochemical staining showed that CD34 positive cells were observed in 2 groups; MVD of experimental group was significantly higher than that of control group at 1 day before pedicle division, and at 3 and 7 days after pedicle division (P lt; 0.05). At 7 days after pedicle division, the VEGF mRNA expression was 7.122 ± 0.503 in experimental group, and was 5.655 ± 1.174 in control group, showing significant difference (t=3.633, P=0.003). Conclusion Natural hirudin can promote the formation of new blood vessels, improve the blood supply of the flap, and increase the survival of random flap after early pedicle division by increasing VEGF expression.

Citation： HAN Zhiqiang,YIN Guoqian,WEI Shuyi,LIN Bojie,PAN Xinyuan.. EFFECT OF NATURAL HIRUDIN ON SURVIVAL OF DORSAL RANDOM FLAP AFTER EARLY PEDICLE DIVISION IN RATS. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(6): 738-742. doi: 10.7507/1002-1892.20130164 Copy

1.	Conforti ML, Connor NP, Heisy DM, et al. Evaluation of performance characteristics of the medicinal leech (Hirudomedicinalis) for the treatment of venous congestion. Plast Reconstr Surg, 2002, 109(1): 228-235.
2.	Tan O, Atik B, Koseoglu B. An easy and safe method to apply leeches on flaps: the leech cage. Plast Reconstr Surg, 2004, 113(1): 466-467.
3.	Ikizceli I, Avsarogullari L, Sözüer E, et al. Bleeding due to a medicinal leech bite. Emerg Med J, 2005, 22(6): 458-460.
4.	Guo-Qian Y, Gang W, Zhi-Yong S. Investigation on the microcirculation effect of local application of natural hirudin on porcine random skin flap venous congestion. Cell Biochem Biophys, 2012, 62(1): 141-146.
5.	林丁盛, 郑鑫, 李志杰, 等. 水蛭素对大鼠随意型皮瓣存活的影响. 中华整形外科杂志, 2011, 27(1): 35-39.
6.	王春艳, 童庭辉, 韩寅, 等. 低分子肝素和重组水蛭素对兔耳静脉淤血皮瓣成活的影响. 中国组织工程研究, 2012, 16(18): 3329-3332.
7.	廖毅, 童庭辉, 王太平, 等. 局部应用重组水蛭素对家兔耳静脉淤血皮瓣成活的影响. 中华烧伤杂志, 2011, 27(3): 215-217.
8.	刘英, 鲁继增, 张松林, 等. 血管内皮生长因子对大鼠皮瓣断蒂时间的影响. 中国临床康复, 2005, 9(22): 97-99.
9.	刘斌, 鲜华, 欧阳晟, 等. bFGF-FG对大鼠随意皮瓣早期断蒂的影响. 国际医药卫生导报, 2009, 15(23): 15-17.
10.	张和韡, 王丽萍. 水蛭素的研究进展. 中国中西医结合肾病杂志, 2013, 14(1): 76-78.
11.	郭应信, 殷国前, 李佳荃, 等. 天然、重组水蛭素对随意皮瓣淤血模型血管内皮细胞生长因子的影响. 中国组织工程研究与临床康复, 2011, 15(7): 1210-1215.
12.	Claesson-Welsh L, Welsh M. VEGFA and tumour angiogenesis. J Internal Medicine, 2013, 273(2): 114-127.
13.	Johnson KE, Wilgus TA. Multiple roles for VEGF in non-melanoma skin cancer: angiogenesis and beyond. Journal of Skin Cancer, 2012, 2012: 483-439.
14.	龚晓燕, 保永亮, 黄金玲. 血管内皮生长因子与肿瘤侵袭转移. 安徽医药, 2013, 17(1): 3-5.
15.	Zhang F, Lineaweaver W. Acute and sustained effects of vascular endothelial growth factor on survival of flaps and skin grafts. Ann Plast Surg, 2011, 66(5): 581-582．.
16.	刘春丽, 刘书会, 刘志辉. 局部应用血管内皮生长因子及骨髓间充质干细胞对超比例随意皮瓣成活率的影响. 华西口腔医学杂志, 2012, 30(6): 645-649.
17.	吴银生, 沈江涌, 姚明, 等. 脂质体介导血管内皮生长因子对不同时间断蒂大鼠随意型皮瓣的影响. 中华整形外科杂志, 2009, 25(2): 120-124.
18.	茹靖涛, 曹靖, 秦涛. pcDNA3.1(+)载体携带血管内皮生长因子治疗大鼠缺血皮瓣. 中华实验外科杂志, 2012, 29(3): 424-427.
19.	Livaoğlu M, Kerimoğlu S, Sönmez B, et al. The effect of Hirudoid on random skin-flap survival in rats. J Plast Reconstr Aesthet Surg, 2010, 63(6): 1047-1051.
20.	Vourtsis SA, Spyriounis PK, Agrogiannis GD, et al. VEGF application on rat skin flap survival. J Invest Surg, 2012, 25(1): 14-19.

1. Conforti ML, Connor NP, Heisy DM, et al. Evaluation of performance characteristics of the medicinal leech (Hirudomedicinalis) for the treatment of venous congestion. Plast Reconstr Surg, 2002, 109(1): 228-235.
2. Tan O, Atik B, Koseoglu B. An easy and safe method to apply leeches on flaps: the leech cage. Plast Reconstr Surg, 2004, 113(1): 466-467.
3. Ikizceli I, Avsarogullari L, Sözüer E, et al. Bleeding due to a medicinal leech bite. Emerg Med J, 2005, 22(6): 458-460.
4. Guo-Qian Y, Gang W, Zhi-Yong S. Investigation on the microcirculation effect of local application of natural hirudin on porcine random skin flap venous congestion. Cell Biochem Biophys, 2012, 62(1): 141-146.
5. 林丁盛, 郑鑫, 李志杰, 等. 水蛭素对大鼠随意型皮瓣存活的影响. 中华整形外科杂志, 2011, 27(1): 35-39.
6. 王春艳, 童庭辉, 韩寅, 等. 低分子肝素和重组水蛭素对兔耳静脉淤血皮瓣成活的影响. 中国组织工程研究, 2012, 16(18): 3329-3332.
7. 廖毅, 童庭辉, 王太平, 等. 局部应用重组水蛭素对家兔耳静脉淤血皮瓣成活的影响. 中华烧伤杂志, 2011, 27(3): 215-217.
8. 刘英, 鲁继增, 张松林, 等. 血管内皮生长因子对大鼠皮瓣断蒂时间的影响. 中国临床康复, 2005, 9(22): 97-99.
9. 刘斌, 鲜华, 欧阳晟, 等. bFGF-FG对大鼠随意皮瓣早期断蒂的影响. 国际医药卫生导报, 2009, 15(23): 15-17.
10. 张和韡, 王丽萍. 水蛭素的研究进展. 中国中西医结合肾病杂志, 2013, 14(1): 76-78.
11. 郭应信, 殷国前, 李佳荃, 等. 天然、重组水蛭素对随意皮瓣淤血模型血管内皮细胞生长因子的影响. 中国组织工程研究与临床康复, 2011, 15(7): 1210-1215.
12. Claesson-Welsh L, Welsh M. VEGFA and tumour angiogenesis. J Internal Medicine, 2013, 273(2): 114-127.
13. Johnson KE, Wilgus TA. Multiple roles for VEGF in non-melanoma skin cancer: angiogenesis and beyond. Journal of Skin Cancer, 2012, 2012: 483-439.
14. 龚晓燕, 保永亮, 黄金玲. 血管内皮生长因子与肿瘤侵袭转移. 安徽医药, 2013, 17(1): 3-5.
15. Zhang F, Lineaweaver W. Acute and sustained effects of vascular endothelial growth factor on survival of flaps and skin grafts. Ann Plast Surg, 2011, 66(5): 581-582．.
16. 刘春丽, 刘书会, 刘志辉. 局部应用血管内皮生长因子及骨髓间充质干细胞对超比例随意皮瓣成活率的影响. 华西口腔医学杂志, 2012, 30(6): 645-649.
17. 吴银生, 沈江涌, 姚明, 等. 脂质体介导血管内皮生长因子对不同时间断蒂大鼠随意型皮瓣的影响. 中华整形外科杂志, 2009, 25(2): 120-124.
18. 茹靖涛, 曹靖, 秦涛. pcDNA3.1(+)载体携带血管内皮生长因子治疗大鼠缺血皮瓣. 中华实验外科杂志, 2012, 29(3): 424-427.
19. Livaoğlu M, Kerimoğlu S, Sönmez B, et al. The effect of Hirudoid on random skin-flap survival in rats. J Plast Reconstr Aesthet Surg, 2010, 63(6): 1047-1051.
20. Vourtsis SA, Spyriounis PK, Agrogiannis GD, et al. VEGF application on rat skin flap survival. J Invest Surg, 2012, 25(1): 14-19.

Chinese Journal of Reparative and Reconstructive Surgery

EFFECT OF NATURAL HIRUDIN ON SURVIVAL OF DORSAL RANDOM FLAP AFTER EARLY PEDICLE DIVISION IN RATS

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