Application of Abdominal Aortic and Bilateral Abdominal Perineal Artery Ligation in Hindlimb Ischemia Model of Rats_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LIU Kun ¹ , YANG Kun ² , LUO Jun ¹ , WANG Yang ¹ , LI Xinxi ¹ , BAI Chao ¹ , JIE Yuegao ¹

1.Center of Digestion and Vascular Surgery, The First Teaching Hospital of Xinjiang Medical University, Urumqi 830054, China;;
2.Department of Burning and Plastic Surgery, The First Teaching Hospital of Xinjiang Medical University, Urumqi 830054, China;

Corresponding author：

Keywords：

Limb ischemia; Abdominal aorta; Abdominal perineal artery; Ligation; Animal model

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Objective To investigate the application of abdominal aorta and bilateral abdominal perineal artery ligation in a hindlimb ischemia model of rats for research.
Methods According to the random digits table, 38 SD rats were divided into 3 groups randomly, sham operation group (SO group, n=12), abdominal aorta ligation group (AAL group, n=12) and abdominal aorta and bilateral abdominal perineal artery ligation group (AAL+BAPA group, n=14). Rats were anesthetized by 6 mg/100 g ketamine via intraperitoneal injection. Blood vessels were ligated via laparotomy according to different procedures. Movement and pale skin color of rat’s hindlimb were observed on 2 and 4 weeks after operation, meanwhile venous blood from unilateral iliac vein was obtained for blood gas analysis and hindlimb skeletal muscle for HE stain.
Results Two rats were dead during 3 d after operation in AAL+BAPA group, other rats survived. Rats in SO group had no obviously abnormal appearance. AAL group and AAL+BAPA group immediately presented hindlimbs pale, lower skin temperature, hypofrontality of limb motion after procedure. Symptoms above mentioned had improved gradually after 2 weeks and completely recuperated 4 weeks after operation in AAL group. Ischemia symptoms were still remained obviously such as cold, dried and thin on the 4th weekend in AAL+BAPA group. Each group had no hindlimbs necrosis. Two weeks after operation, pale limbs and muscle strength in AAL+BAPA group were more severe than those of SO group (P lt;0.05); Pale limbs was still worse than that of AAL group on 4 weeks after operation (P lt;0.05). There were no significant differences on different time in each group (P gt;0.05). Venous blood partial pressure of oxygen of AAL+PABA group was significantly lower than that of the other two groups on 2 and 4 weeks after operation (P lt;0.05). Normal striated muscle structure was presented in SO group pathologically. AAL group revealed coloretur unevenness, swelling and distension, muscle cellular transverse striation elimination, skeletal muscle cell nucleolus deeply stained on the 2nd weekend and no difference with the SO group on the 4th weekend. AAL+BAPA group presented skeletal muscles decoration unevenness, cells swelling and distension, muscle cellular transverse striation elimination, skeletal muscle cell nucleolus stained deeply and intensively, intercellular space widening until the 4th weekend， but no obviously necrocytosis.
Conclusion The method of ligating abdominal aorta and bilateral abdominal perineal artery can make a stable SD rat model of hindlimb ischemia

Citation： LIU Kun ,YANG Kun,LUO Jun,WANG Yang,LI Xinxi,BAI Chao,JIE Yuegao. Application of Abdominal Aortic and Bilateral Abdominal Perineal Artery Ligation in Hindlimb Ischemia Model of Rats. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2010, 17(3): 258-262. doi: Copy

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11.	Hiasa K, Ishibashi M, Ohtani K, et al. Gene transfer of stromal cellderived factor1α enhances ischemic vasculogenesis and angiogenesis via vascular endothelial growth factor/endothelial nitric oxide synthaserelated pathway: nextgeneration chemokine therapy for therapeutic neovascularization [J]. Circulation, 2004; 109(20): 24542461.
12.	Sato Y, Matsui K, Ajiki T, et al. Can a bone marrow cell contribute to organ regeneration? In vivo analysis using transgenic rats with reporter genes [J]. Transplant Proc, 2005; 37(1): 273275.
13.	Otsuka H, Akashi H, Murohara T, et al. The prostacyclin analog beraprost sodium augments the efficacy of therapeutic angiogenesis induced by autologous bone marrow cells [J]. Ann Vasc Surg, 2006; 20(5): 646652.
14.	de Nigris F, Balestrieri ML, WilliamsIgnarro S, et al. Therapeutic effects of autologous bone marrow cells and metabolic intervention in the ischemic hindlimb of spontaneously hypertensive rats involve reduced cell senescence and CXCR4/Akt/eNOS pathways [J]. J Cardiovasc Pharmacol, 2007; 50(4): 424433.
15.	Qin SL, Li TS, Kubo M, et al. Transient increase of cytokines in the acute ischemic tissue is beneficial to cellbased therapeutic angiogenesis [J]. Circ J, 2008; 72(12): 20752080.
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17.	Iohara K, Zheng L, Wake H, et al. A novel stem cell source for vasculogenesis in ischemia: subfraction of side population cells from dental pulp [J]. Stem Cells, 2008; 26(9): 24082418.
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25.	Iwatsuki Y, Sakata C, Kawasaki T, et al. Experimental model of lower limb ischemia in rats and the effect of YM466, an oral direct factor Xa inhibitor [J]. Biol Pharm Bull, 2007; 30(10): 18741877.
26.	梁翠宏，田铧，徐蕴，等. 结扎切断法与白芨微粒栓塞法建立大鼠后肢缺血模型效果比较 [J]. 山东大学学报(医学版), 2007; 45(10): 10081010.
27.	Pu LQ, Jackson S, Lachapelle KJ, et al. A persistent hindlimb ischemia model in the rabbit [J]. J Invest Surg, 1994; 7(1): 4960.
28.	梁翠宏，田铧，徐蕴，等. 结扎切断法制作大鼠后肢缺血模型的效果 [J]. 解剖学杂志, 2006; 29(6): 795797.
29.	杨安峰, 王平. 大鼠的解剖和组织 [M]. 北京: 科学出版社, 1980: 124130.

1. Isner JM, Walsh K, Symes J, et al. Arterial gene transfer for therapeutic angiogenesis in patients with peripheral artery disease [J]. Hum Gene Ther, 1996; 7(8): 959988.
2. Tatsumi T, Matsubara H. Therapeutic angiogenesis for peripheral arterial disease and ischemic heart disease by autologous bone marrow cells implantation [J]. Nippon Rinsho, 2006; 64(11): 21262134.
3. Sumi M, Sata M, Toya N, et al. Transplantation of adipose stromal cells, but not mature adipocytes, augments ischemiainduced angiogenesis [J]. Life Sci, 2007; 80(6): 559565.
4. Hamano K, Li TS, Kobayashi T, et al. The induction of angiogenesis by the implantation of autologous bone marrow cells: a novel and simple therapeutic method [J]. Surgery, 2001; 130(1): 4454.
5. Ikenaga S, Hamano K, Nishida M, et al. Autologous bone marrow implantation induced angiogenesis and improved deteriorated exercise capacity in a rat ischemic hindlimb model [J]. J Surg Res, 2001; 96(2): 277283.
6. Kawamoto A, Asahara T, Losordo DW. Transplantation of endothelial progenitor cells for therapeutic neovascularization [J]. Cardiovasc Radiat Med, 2002; 3(34): 221225.
7. Iba O, Matsubara H, Nozawa Y, et al. Angiogenesis by implantation of peripheral blood mononuclear cells and platelets into ischemic limbs [J]. Circulation, 2002; 106(15): 20192025.
8. Iwaguro H, Yamaguchi J, Kalka C, et al. Endothelial progenitor cell vascular endothelial growth factor gene transfer for vascular regeneration [J]. Circulation, 2002; 105(6): 732738.
9. Padilla L, Krtzsch E, Schalch P, et al. Administration of bone marrow cells into surgically induced fibrocollagenous tunnels induces angiogenesis in ischemic rat hindlimb model [J]. Microsurgery, 2003; 23(6): 568574.
10. 刘文亮, 尹邦良, 喻风雷, 等. 骨髓单个核细胞缺血组织局部移植对血运重建的影响 [J]. 中南大学学报(医学版), 2004; 29(5): 572576.
11. Hiasa K, Ishibashi M, Ohtani K, et al. Gene transfer of stromal cellderived factor1α enhances ischemic vasculogenesis and angiogenesis via vascular endothelial growth factor/endothelial nitric oxide synthaserelated pathway: nextgeneration chemokine therapy for therapeutic neovascularization [J]. Circulation, 2004; 109(20): 24542461.
12. Sato Y, Matsui K, Ajiki T, et al. Can a bone marrow cell contribute to organ regeneration? In vivo analysis using transgenic rats with reporter genes [J]. Transplant Proc, 2005; 37(1): 273275.
13. Otsuka H, Akashi H, Murohara T, et al. The prostacyclin analog beraprost sodium augments the efficacy of therapeutic angiogenesis induced by autologous bone marrow cells [J]. Ann Vasc Surg, 2006; 20(5): 646652.
14. de Nigris F, Balestrieri ML, WilliamsIgnarro S, et al. Therapeutic effects of autologous bone marrow cells and metabolic intervention in the ischemic hindlimb of spontaneously hypertensive rats involve reduced cell senescence and CXCR4/Akt/eNOS pathways [J]. J Cardiovasc Pharmacol, 2007; 50(4): 424433.
15. Qin SL, Li TS, Kubo M, et al. Transient increase of cytokines in the acute ischemic tissue is beneficial to cellbased therapeutic angiogenesis [J]. Circ J, 2008; 72(12): 20752080.
16. Wragg A, Mellad JA, Beltran LE, et al. VEGFR1/CXCR4positive progenitor cells modulate local inflammation and augment tissue perfusion by a SDF1dependent mechanism [J]. J Mol Med, 2008; 86(11): 12211232.
17. Iohara K, Zheng L, Wake H, et al. A novel stem cell source for vasculogenesis in ischemia: subfraction of side population cells from dental pulp [J]. Stem Cells, 2008; 26(9): 24082418.
18. Tajima H, IwaiTakano M, Yaoita H, et al. Mast cells contribute to flow restoration by bone marrow cell transplantation in rats with ischemic limbs [J]. Int Heart J, 2009; 50(2): 247257.
19. 汪忠镐, 李震. 我国血管外科的现状和发展 [J]. 中国普外基础与临床杂志, 2006; 13(6): 625628.
20. 梁翠宏. Wistar大鼠后肢缺血模型的实验研究 [D]. 山东大学, 2005: 17.
21. 刘万里, 薛茜, 曹明芹，等. 用 SPSS实现完全随机设计多组比较秩和检验的多重比较 [J]. 地方病通报, 2007; 22(2): 2729.
22. 崔世军，俞恒锡，滕旭，等. 家兔后肢缺血模型的制作 [J]. 中国比较医学杂志, 2003; 13(1): 3638.
23. Heil M, Ziegelhoeffer T, Pipp F, et al. Blood monocyte concentration is critical for enhancement of collateral artery growth [J]. Am J Physiol Heart Circ Physiol, 2002; 283(6): H2411H2419.
24. 霍鑫，冯金炜，刘方峰，等. pEGFPC1/Akt体外转染骨髓间充质干细胞对后肢缺血大鼠血管生成的影响 [J]. 中国普外基础与临床杂志, 2008; 15(6): 402407.
25. Iwatsuki Y, Sakata C, Kawasaki T, et al. Experimental model of lower limb ischemia in rats and the effect of YM466, an oral direct factor Xa inhibitor [J]. Biol Pharm Bull, 2007; 30(10): 18741877.
26. 梁翠宏，田铧，徐蕴，等. 结扎切断法与白芨微粒栓塞法建立大鼠后肢缺血模型效果比较 [J]. 山东大学学报(医学版), 2007; 45(10): 10081010.
27. Pu LQ, Jackson S, Lachapelle KJ, et al. A persistent hindlimb ischemia model in the rabbit [J]. J Invest Surg, 1994; 7(1): 4960.
28. 梁翠宏，田铧，徐蕴，等. 结扎切断法制作大鼠后肢缺血模型的效果 [J]. 解剖学杂志, 2006; 29(6): 795797.
29. 杨安峰, 王平. 大鼠的解剖和组织 [M]. 北京: 科学出版社, 1980: 124130.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Application of Abdominal Aortic and Bilateral Abdominal Perineal Artery Ligation in Hindlimb Ischemia Model of Rats

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