• 武漢大學(xué)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(jìn)行評價(jià),為后續(xù)的干預(yù)實(shí)驗(yàn)提供研究平臺(tái)。
方法  將15只SD大鼠隨機(jī)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(dòng)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(dòng)脈,不予結(jié)扎。2周后對3組大鼠股動(dòng)脈的起始段行彩色多普勒超聲檢查,以檢測股動(dòng)脈的血流峰值速度和血流加速時(shí)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(xì)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(shí)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(dòng)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(shí)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(xì)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(xì)血管密度〔(1.40±0.55) 個(gè)/HPF〕小于正常對照組 〔(6.80±0.84) 個(gè)/HPF〕及糖尿病組〔(4.60±0.55)個(gè)/HPF〕,差異均有統(tǒng)計(jì)學(xué)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(dǎo)2型糖尿病模型,在此模型基礎(chǔ)上結(jié)扎髂總動(dòng)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(jià)△. 中國普外基礎(chǔ)與臨床雜志, 2013, 20(12): 1358-1362. doi: 復(fù)制

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(jià)[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(dǎo)大鼠2型糖尿病模型的建立[J]. 吉林醫(yī)藥學(xué)院學(xué)報(bào), 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(jià)兔骨髓間充質(zhì)干細(xì)胞自體移植對缺血肢體血管再生的影響[J]. 中華醫(yī)學(xué)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(xué), 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(lán)榮芳, 等. AAV介導(dǎo)的CD151基因促大鼠缺血后肢血管再生和血運(yùn)重建的實(shí)驗(yàn)研究[J]. 華中科技大學(xué)學(xué)報(bào):醫(yī)學(xué)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(xì)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(qiáng), 王薇, 等. 前列地爾脂微球載體注射液對糖尿病下肢動(dòng)脈血管病變的影響[J]. 中國實(shí)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(shí)驗(yàn)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(dòng)脈結(jié)扎建立有效的大鼠后肢缺血模型[J]. 解剖學(xué)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(jià)[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(dǎo)大鼠2型糖尿病模型的建立[J]. 吉林醫(yī)藥學(xué)院學(xué)報(bào), 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(jià)兔骨髓間充質(zhì)干細(xì)胞自體移植對缺血肢體血管再生的影響[J]. 中華醫(yī)學(xué)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(xué), 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(lán)榮芳, 等. AAV介導(dǎo)的CD151基因促大鼠缺血后肢血管再生和血運(yùn)重建的實(shí)驗(yàn)研究[J]. 華中科技大學(xué)學(xué)報(bào):醫(yī)學(xué)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(xì)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(qiáng), 王薇, 等. 前列地爾脂微球載體注射液對糖尿病下肢動(dòng)脈血管病變的影響[J]. 中國實(shí)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(shí)驗(yàn)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(dòng)脈結(jié)扎建立有效的大鼠后肢缺血模型[J]. 解剖學(xué)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.