• 武漢大學人民醫(yī)院內(nèi)分泌科(湖北武漢 430060);

目的  建立2型糖尿病大鼠后肢缺血模型并進行評價,為后續(xù)的干預(yù)實驗提供研究平臺。
方法  將15只SD大鼠隨機分為正常對照組、糖尿病組及糖尿病后肢缺血組,每組5只。糖尿病組及糖尿病后肢缺血組的10只大鼠均給予高脂飲食喂養(yǎng)4周后,腹腔注射鏈脲佐菌素(STZ,40 mg/kg)以建立2型糖尿病模型。糖尿病后肢缺血組大鼠建模成功后行雙側(cè)髂總動脈結(jié)扎術(shù)以建立后肢缺血模型,正常對照組和糖尿病組大鼠僅分離髂總動脈,不予結(jié)扎。2周后對3組大鼠股動脈的起始段行彩色多普勒超聲檢查,以檢測股動脈的血流峰值速度和血流加速時間;取缺血部位的小腿三頭肌及大腿股四頭肌組織,分別行HE染色及免疫組化SP染色,以觀察3組大鼠肌細胞的營養(yǎng)狀況及血管再生情況。
結(jié)果  后肢缺血模型建模2周后,正常對照組、糖尿病組和糖尿病后肢缺血組大鼠的血流峰值速度分別為(22.49±3.02) cm/s、(17.36±2.60) cm/s和(11.23±1.26) cm/s,血流加速時間分別為(0.080±0.009) s、(0.120±0.009) s和(0.160±0.020) s,糖尿病后肢缺血組大鼠的股動脈血流峰值速度小于正常對照組和糖尿病組(P<0.05),而血流加速時間較長(P<0.05)。HE染色結(jié)果顯示:糖尿病后肢缺血組大鼠小腿三頭肌的結(jié)構(gòu)破壞,有大量炎癥細胞浸潤,肌肉損傷程度重于正常對照組和糖尿病組。免疫組化SP染色結(jié)果顯示:糖尿病后肢缺血組大鼠大腿股四頭肌的毛細血管密度〔(1.40±0.55) 個/HPF〕小于正常對照組 〔(6.80±0.84) 個/HPF〕及糖尿病組〔(4.60±0.55)個/HPF〕,差異均有統(tǒng)計學意義(P<0.05)。
結(jié)論  對SD大鼠給予高脂飲食聯(lián)合小劑量STZ注射可以成功誘導2型糖尿病模型,在此模型基礎(chǔ)上結(jié)扎髂總動脈可以成功制備糖尿病后肢缺血模型。

引用本文: 余愛林,李競,高凌. 2型糖尿病大鼠后肢缺血模型的建立及評價△. 中國普外基礎(chǔ)與臨床雜志, 2013, 20(12): 1358-1362. doi: 復(fù)制

版權(quán)信息: ?四川大學華西醫(yī)院華西期刊社《中國普外基礎(chǔ)與臨床雜志》版權(quán)所有,未經(jīng)授權(quán)不得轉(zhuǎn)載、改編

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. 楊盛家, 陳兵, 羅濤, 等. Lewis大鼠慢性后肢缺血模型的制備與評價[J]. 中國普外基礎(chǔ)與臨床雜志, 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. 王春艷. 鏈脲佐菌素誘導大鼠2型糖尿病模型的建立[J]. 吉林醫(yī)藥學院學報, 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. 朱棟曉, 苗宗寧, 錢寒光, 等. 超聲評價兔骨髓間充質(zhì)干細胞自體移植對缺血肢體血管再生的影響[J]. 中華醫(yī)學超聲雜志:電子版, 2008, 5(2):223-228.
11. 白雪琴, 韓萍. 代謝綜合征患病率的調(diào)查及健康管理方案研究[J]. 中國療養(yǎng)醫(yī)學, 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. 康基順, 劉正湘, 藍榮芳, 等. AAV介導的CD151基因促大鼠缺血后肢血管再生和血運重建的實驗研究[J]. 華中科技大學學報:醫(yī)學版, 2006, 35(5):605-608.
14. 霍鑫, 馮金煒, 劉方峰, 等. pEGFP-C1/Akt體外轉(zhuǎn)染骨髓間充質(zhì)干細胞對后肢缺血大鼠血管生成的影響[J]. 中國普外基礎(chǔ)與臨床雜志, 2008, 15(6):402-407.
15. 李靜宜, 李強, 王薇, 等. 前列地爾脂微球載體注射液對糖尿病下肢動脈血管病變的影響[J]. 中國實用內(nèi)科雜志, 2007, 27(8):597-599.
16. 劉程偉, 張兆寰, 張雪松, 等. 重組人生長激素促大鼠缺血后肢血管再生的實驗研究[J]. 中華臨床醫(yī)師雜志:電子版, 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. 鄒知耕, 田鏵, 趙世剛, 等. 自體血栓并股動脈結(jié)扎建立有效的大鼠后肢缺血模型[J]. 解剖學雜志, 2011, 34(5):718-720.
26. 楊盛家, 陳兵, 羅濤, 等. 大鼠后肢急性缺血模型的構(gòu)建及評估[J]. 中國普通外科雜志, 2009, 18(6):580-583.
  1. 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. 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. 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. 4. 楊盛家, 陳兵, 羅濤, 等. Lewis大鼠慢性后肢缺血模型的制備與評價[J]. 中國普外基礎(chǔ)與臨床雜志, 2012, 19(12):1291-1297.
  5. 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. 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. 7. 王春艷. 鏈脲佐菌素誘導大鼠2型糖尿病模型的建立[J]. 吉林醫(yī)藥學院學報, 2010, 31(1):22-23.
  8. 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. 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. 10. 朱棟曉, 苗宗寧, 錢寒光, 等. 超聲評價兔骨髓間充質(zhì)干細胞自體移植對缺血肢體血管再生的影響[J]. 中華醫(yī)學超聲雜志:電子版, 2008, 5(2):223-228.
  11. 11. 白雪琴, 韓萍. 代謝綜合征患病率的調(diào)查及健康管理方案研究[J]. 中國療養(yǎng)醫(yī)學, 2011, 20(1):10-12.
  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. 13. 康基順, 劉正湘, 藍榮芳, 等. AAV介導的CD151基因促大鼠缺血后肢血管再生和血運重建的實驗研究[J]. 華中科技大學學報:醫(yī)學版, 2006, 35(5):605-608.
  14. 14. 霍鑫, 馮金煒, 劉方峰, 等. pEGFP-C1/Akt體外轉(zhuǎn)染骨髓間充質(zhì)干細胞對后肢缺血大鼠血管生成的影響[J]. 中國普外基礎(chǔ)與臨床雜志, 2008, 15(6):402-407.
  15. 15. 李靜宜, 李強, 王薇, 等. 前列地爾脂微球載體注射液對糖尿病下肢動脈血管病變的影響[J]. 中國實用內(nèi)科雜志, 2007, 27(8):597-599.
  16. 16. 劉程偉, 張兆寰, 張雪松, 等. 重組人生長激素促大鼠缺血后肢血管再生的實驗研究[J]. 中華臨床醫(yī)師雜志:電子版, 2011, 5(11):3146-3150.
  17. 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. 18. Kalka C, Baumgartner I. Gene and stem cell therapy in peripheralarterial occlusive disease[J]. Vasc Med, 2008, 13(2):157-172.
  19. 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. 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. 21. Lachmann N, Nikol S. Therapeutic angiogenesis for peripheral artery disease:stem cell therapy[J]. Vasa, 2007, 36(4):241-251.
  22. 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. 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. 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. 25. 鄒知耕, 田鏵, 趙世剛, 等. 自體血栓并股動脈結(jié)扎建立有效的大鼠后肢缺血模型[J]. 解剖學雜志, 2011, 34(5):718-720.
  26. 26. 楊盛家, 陳兵, 羅濤, 等. 大鼠后肢急性缺血模型的構(gòu)建及評估[J]. 中國普通外科雜志, 2009, 18(6):580-583.