Establishment and Evaluation of Ischemic Hind Limb Model in Type 2 Diabetic Rats_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

YU Ailin ,  LI Jing , GAO Ling.

Department of Endocrinology， Renmin Hospital， Wuhan University， Wuhan 430060， Hubei Province， China;

Corresponding author：

LI Jing, Email: lijing7823@hotmail.com

Keywords：

Type 2 diabetes; Ischemic hind limb model; Rat; Evaluation

Video：

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Objective 　To establish and evaluate the hind limb ischemia model in type 2 diabetic rats， and to provide
a platform for subsequent intervention experiment.
Methods 　Fifteen SD rats were randomly divided into 3 groups：normal control group， diabetes group， and diabetic ischemia group， each group enrolled 5 rats. The 10 rats of later 2 groups were fed with high fat diet for 4 weeks， and then received intraperitoneal injection of streptozotocin （40 mg/kg） to establish type 2 diabetic model. Three days later， the rats of diabetic ischemia group underwent ligation of the bilateral common iliac artery to establish the hind limb ischemia model， but iliac artery of rats in other 2 groups didn’t received ligation. Ultrasound and color Doppler flow imaging detection was used to determine the peak velocity and acceleration time of femoral artery in rats of 3 groups in 2 weeks after operation， and triceps tissues and quadriceps tissues were collected to perform HE staining and SP staining for the observation on status of nutrition and vascular regeneration respectively.
Results 　The peak velocity of femoral artery in rats of normal control group， diabetes group， and diabetic ischemia group were （22.49±3.02） cm/s，（17.36±2.60） cm/s， and （11.23±1.26） cm/s， the acceleration time were （0.080±0.009） s，（0.120±0.009）s， and（0.160±0.020） s， the arteriolar density measured by immunohistochemistry SP method were 6.80±0.84/HPF， 4.60±0.55/HPF， and 1.40±0.55/HPF respectively. The peak velocity of femoral artery and arteriolar density of rats in diabetic ischemia group were both lower （P＜0.05）， but acceleration time was longer （P＜0.05）. Results of HE staining showed that structure of triceps tissues was damaged， with infiltration of lots of inflamm-atory cells， which was worse than normal control group and diabetes group.
Conclusion 　Method of high-fat diet in combination with small dose of streptozotocin can induce type 2 diabetic rat model， and hind limb ischemia model can be successfully established by ligation of common iliac artery on this model.

Citation： YU Ailin,LI Jing,GAO Ling.. Establishment and Evaluation of Ischemic Hind Limb Model in Type 2 Diabetic Rats. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2013, 20(12): 1358-1362. doi: Copy

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10.	朱栋晓，苗宗宁，钱寒光，等. 超声评价兔骨髓间充质干细胞自体移植对缺血肢体血管再生的影响[J]. 中华医学超声杂志：电子版， 2008， 5(2)：223-228.
11.	白雪琴，韩萍. 代谢综合征患病率的调查及健康管理方案研究[J]. 中国疗养医学， 2011， 20(1)：10-12.
12.	de Paula EV， Flores-Nascimento MC， Arruda VR， et al. Dual gene transfer of fibroblast growth factor-2 and platelet derived growth factor-BB using plasmid deoxyribonucleic acid promotes effective angiogenesis and arteriogenesis in a rodent model of hindlimb ischemia[J]. Transl Res， 2009， 153(5)：232-239.
13.	康基顺，刘正湘，蓝荣芳，等. AAV介导的CD151基因促大鼠缺血后肢血管再生和血运重建的实验研究[J]. 华中科技大学学报：医学版， 2006， 35(5)：605-608.
14.	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.
15.	李静宜，李强，王薇，等. 前列地尔脂微球载体注射液对糖尿病下肢动脉血管病变的影响[J]. 中国实用内科杂志， 2007， 27(8)：597-599.
16.	刘程伟，张兆寰，张雪松，等. 重组人生长激素促大鼠缺血后肢血管再生的实验研究[J]. 中华临床医师杂志：电子版， 2011， 5(11)：3146-3150.
17.	Burghelea C， Lucan M， Ghervan L， et al. Advantages of ilio-inguinal extraction of the kidney after retroperitoneoscopic nephroureterectomy[J]. Chirurgia (Bucur)， 2008， 103(1)：61-66.
18.	Kalka C， Baumgartner I. Gene and stem cell therapy in peripheralarterial occlusive disease[J]. Vasc Med， 2008， 13(2)：157-172.
19.	Hudlická O， Garnham A， Shiner R， et al. Attenuation of changes in capillary fine structure and leukocyte adhesion improves muscle performance following chronic ischaemia in rats[J]. J Physiol， 2008， 586(Pt 20)：4961-4975.
20.	Nikol S. Herapeutic angiogenesis using gene transfer and stem cell therapy in peripheral artery disease[J]. Dtsch Med Wochenschr， 2011， 136(14)：672-674.
21.	Lachmann N， Nikol S. Therapeutic angiogenesis for peripheral artery disease：stem cell therapy[J]. Vasa， 2007， 36(4)：241-251.
22.	Lian Q， Zhang Y， Zhang J， et al. Functional mesenchymal stem cells derived from human induced pluripotent stem cells attenuate limb ischemia in mice[J]. Circulation， 2010， 121(9)：1113-1123.
23.	Staudacher DL， Sela Y， Itskovitz-Eldor J， et al. Intra-arterial injection of human embryonic stem cells in athymic rat hind limb ischemia model leads to arteriogenesis[J]. Cardiovasc Revasc Med， 2011， 12(4)：228-234.
24.	Nagano A， Komatsuno T， Kasahara H， et al. Evaluation of hepatocyte growth factor plasmid therapeutic effect by 99Tcm-hexakis-2-methyoxy-isobutylisonitrile blood flow scintigraphy in a rat model of hind limb ischemia[J]. Nucl Med Commun， 2011， 32(9)：818-823.
25.	邹知耕，田铧，赵世刚，等. 自体血栓并股动脉结扎建立有效的大鼠后肢缺血模型[J]. 解剖学杂志， 2011， 34(5)：718-720.
26.	杨盛家，陈兵，罗涛，等. 大鼠后肢急性缺血模型的构建及评估[J]. 中国普通外科杂志， 2009， 18(6)：580-583.

1. Ishak NA， Ismail M， Hamid M， et al. Antidiabetic and hypolipidemic activities of curculigo latifolia fruit：root extract in high fat fed diet and low dose STZ induced diabetic rats[J]. Evid Based Complement Alternat Med， 2013， 2013：601838.
2. Bas AL， Demirci S， Yazihan N， et al. Nerium oleander distillate improves fat and glucose metabolism in high-fat diet-fed streptozotocin-induced diabetic rats[J]. Int J Endocrinol， 2012， 2012：947187.
3. Davidson EP， Coppey LJ， Holmes A， et al. Effect of treatment of high fat fed/low dose streptozotocin-diabetic rats with ilepatril on vascular and neural complications[J]. Eur J Pharmacol， 2011， 668(3)：497-506.
4. 杨盛家，陈兵，罗涛，等. Lewis大鼠慢性后肢缺血模型的制备与评价[J]. 中国普外基础与临床杂志， 2012， 19(12)：1291-1297.
5. Yang Z， von Ballmoos MW， Diehm N， et al. Call for a reference model of chronic hind limb ischemia to investigate therapeutic angiogenesis[J]. Vascul Pharmacol， 2009， 51(4)：268-274.
6. Zhang Z， Xue HL， Liu Y， et al. Yi-Qi-Zeng-Min-Tang， a Chin-ese medicine， ameliorates insulin resistance in type 2 diabetic rats[J]. World J Gastroenterol， 2011， 17(8)：987-995.
7. 王春艳. 链脲佐菌素诱导大鼠2型糖尿病模型的建立[J]. 吉林医药学院学报， 2010， 31(1)：22-23.
8. Huang AL， Silver AE， Shvenke E， et al. Predictive value of reac-tive hyperemia for cardiovascular events in patients with peripheralarterial disease undergoing vascular surgery[J]. Arterioscler Thromb Vasc Biol， 2007， 27(10)：2113-2119.
9. Corcoran HA， Smith BE， Mathers P， et al. Laser doppler imagingof reactive hyperemia exposes blood flow deficits in a rat model of experimental limb ischemia[J]. J Cardiovasc Pharmacol， 2009， 53(6)：446-451.
10. 朱栋晓，苗宗宁，钱寒光，等. 超声评价兔骨髓间充质干细胞自体移植对缺血肢体血管再生的影响[J]. 中华医学超声杂志：电子版， 2008， 5(2)：223-228.
11. 白雪琴，韩萍. 代谢综合征患病率的调查及健康管理方案研究[J]. 中国疗养医学， 2011， 20(1)：10-12.
12. de Paula EV， Flores-Nascimento MC， Arruda VR， et al. Dual gene transfer of fibroblast growth factor-2 and platelet derived growth factor-BB using plasmid deoxyribonucleic acid promotes effective angiogenesis and arteriogenesis in a rodent model of hindlimb ischemia[J]. Transl Res， 2009， 153(5)：232-239.
13. 康基顺，刘正湘，蓝荣芳，等. AAV介导的CD151基因促大鼠缺血后肢血管再生和血运重建的实验研究[J]. 华中科技大学学报：医学版， 2006， 35(5)：605-608.
14. 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.
15. 李静宜，李强，王薇，等. 前列地尔脂微球载体注射液对糖尿病下肢动脉血管病变的影响[J]. 中国实用内科杂志， 2007， 27(8)：597-599.
16. 刘程伟，张兆寰，张雪松，等. 重组人生长激素促大鼠缺血后肢血管再生的实验研究[J]. 中华临床医师杂志：电子版， 2011， 5(11)：3146-3150.
17. Burghelea C， Lucan M， Ghervan L， et al. Advantages of ilio-inguinal extraction of the kidney after retroperitoneoscopic nephroureterectomy[J]. Chirurgia (Bucur)， 2008， 103(1)：61-66.
18. Kalka C， Baumgartner I. Gene and stem cell therapy in peripheralarterial occlusive disease[J]. Vasc Med， 2008， 13(2)：157-172.
19. Hudlická O， Garnham A， Shiner R， et al. Attenuation of changes in capillary fine structure and leukocyte adhesion improves muscle performance following chronic ischaemia in rats[J]. J Physiol， 2008， 586(Pt 20)：4961-4975.
20. Nikol S. Herapeutic angiogenesis using gene transfer and stem cell therapy in peripheral artery disease[J]. Dtsch Med Wochenschr， 2011， 136(14)：672-674.
21. Lachmann N， Nikol S. Therapeutic angiogenesis for peripheral artery disease：stem cell therapy[J]. Vasa， 2007， 36(4)：241-251.
22. Lian Q， Zhang Y， Zhang J， et al. Functional mesenchymal stem cells derived from human induced pluripotent stem cells attenuate limb ischemia in mice[J]. Circulation， 2010， 121(9)：1113-1123.
23. Staudacher DL， Sela Y， Itskovitz-Eldor J， et al. Intra-arterial injection of human embryonic stem cells in athymic rat hind limb ischemia model leads to arteriogenesis[J]. Cardiovasc Revasc Med， 2011， 12(4)：228-234.
24. Nagano A， Komatsuno T， Kasahara H， et al. Evaluation of hepatocyte growth factor plasmid therapeutic effect by 99Tcm-hexakis-2-methyoxy-isobutylisonitrile blood flow scintigraphy in a rat model of hind limb ischemia[J]. Nucl Med Commun， 2011， 32(9)：818-823.
25. 邹知耕，田铧，赵世刚，等. 自体血栓并股动脉结扎建立有效的大鼠后肢缺血模型[J]. 解剖学杂志， 2011， 34(5)：718-720.
26. 杨盛家，陈兵，罗涛，等. 大鼠后肢急性缺血模型的构建及评估[J]. 中国普通外科杂志， 2009， 18(6)：580-583.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Establishment and Evaluation of Ischemic Hind Limb Model in Type 2 Diabetic Rats

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