高脂饮食与线栓法制作慢性缺血模型鼠:方法的评估与比较
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摘要
背景:有研究使用线栓法建立慢性缺血动物模型,但其观察周期较长,实验动物经过手术及疾病的打击,生存期有可能会缩短。作者在此基础上加用高脂饮食因素,进一步确立可靠的慢性缺血动物模型。目的:探讨高脂饮食大鼠线栓法建立慢性缺血动物模型的可行性。方法:健康2周龄SD大鼠40只,体质量为50-70g,由承德医学院动物实验中心提供。采用随机数字表法将大鼠分成实验组及对照组:实验组大鼠选用高脂饲料进行喂养,对照组大鼠选用普通饲料进行喂养,2组均采用线栓法建立大鼠后肢缺血模型。分别于大鼠模型建立成功后第1,2,4,6周进行肌力观察、激光多普勒血流仪检测后肢血流情况、苏木精-伊红染色观察腓肠肌组织和血管组织变化。结果及结论:(1)实验组造模后1周大部分大鼠出现肌力异常表现,对照组大部分未出现(P <0.05);造模后2周及4周2组大鼠大部分均出现明显肌力异常表现,差异无显著性意义;造模后6周对照组肌力异常例数少于实验组;(2)激光多普勒血流检测及组织学检测均提示实验组较对照组要早出现缺血表现,且实验组缺血表现恢复程度较对照组要低;(3)结果说明,线栓法可有效制作慢性缺血大鼠模型,合并高脂饮食可较普通饮食早出现缺血表现,且高脂饮食可使慢性缺血状态持续时间更长。
BACKGROUND: Some studies have prepared a chronic ischemic animal model using the thread embolism technique, but the observation period is longer, and the survival time of the experimental animals may be shortened because of surgical trauma and disease. Herein, the authors further established a reliable animal model of chronic ischemia using the thread embolism technique by addition of high-fat diet. OBJECTIVE: To investigate the feasibility of establishing a reliable animal model of chronic ischemia in high-fat diet rats using the thread embolism method.METHODS: Forty Sprague-Dawley rats, 2 weeks of age, weighing 50-70 g, were provided by the Animal Experimental Center of Chengde Medical University in China. The rats were randomly divided into two groups, and fed with high-fat diet(experimental group) and normal diet(control group), respectively. Chronic ischemia animal models were then established in the hind limbs of all the rats using the thread embolism technique. Muscle strength of the rats was detected at 1, 2, 4, and 6 weeks postoperatively. Meanwhile, blood flow changes in the hind limbs and histological changes of the gastrocnemius and vascular tissue were observed using laser Doppler and hematoxylin-eosin staining, respectively. RESULTS AND CONCLUSION:(1) Most of the rats in the experimental group had abnormal muscle performance at 1 week postoperatively. Most of the rats in the control group had nothing-unusual(P < 0.05). Most of the rats in both groups appeared with abnormal muscle performance at 2 and 4 weeks postoperatively, and there was no significant difference between the two groups. At 6 weeks postoperatively, the number of rats with abnormal muscle performance was lower in the control group than the experimental group.(2) Laser Doppler flow examination and histological examination both showed that the experimental group had ischemic symptoms earlier than the control group, and worse recovery from ischemia was found in the experimental group compared to the control group. These findings indicate that the thread embolism method can effectively make a rat model of chronic ischemia, and the addition of high-fat diet can cause an ischemic appearance earlier than the addition of normal diet, indicating the high-fat diet can prolong the state of chronic ischemia.
BACKGROUND: Some studies have prepared a chronic ischemic animal model using the thread embolism technique, but the observation period is longer, and the survival time of the experimental animals may be shortened because of surgical trauma and disease. Herein, the authors further established a reliable animal model of chronic ischemia using the thread embolism technique by addition of high-fat diet. OBJECTIVE: To investigate the feasibility of establishing a reliable animal model of chronic ischemia in high-fat diet rats using the thread embolism method.METHODS: Forty Sprague-Dawley rats, 2 weeks of age, weighing 50-70 g, were provided by the Animal Experimental Center of Chengde Medical University in China. The rats were randomly divided into two groups, and fed with high-fat diet(experimental group) and normal diet(control group), respectively. Chronic ischemia animal models were then established in the hind limbs of all the rats using the thread embolism technique. Muscle strength of the rats was detected at 1, 2, 4, and 6 weeks postoperatively. Meanwhile, blood flow changes in the hind limbs and histological changes of the gastrocnemius and vascular tissue were observed using laser Doppler and hematoxylin-eosin staining, respectively. RESULTS AND CONCLUSION:(1) Most of the rats in the experimental group had abnormal muscle performance at 1 week postoperatively. Most of the rats in the control group had nothing-unusual(P < 0.05). Most of the rats in both groups appeared with abnormal muscle performance at 2 and 4 weeks postoperatively, and there was no significant difference between the two groups. At 6 weeks postoperatively, the number of rats with abnormal muscle performance was lower in the control group than the experimental group.(2) Laser Doppler flow examination and histological examination both showed that the experimental group had ischemic symptoms earlier than the control group, and worse recovery from ischemia was found in the experimental group compared to the control group. These findings indicate that the thread embolism method can effectively make a rat model of chronic ischemia, and the addition of high-fat diet can cause an ischemic appearance earlier than the addition of normal diet, indicating the high-fat diet can prolong the state of chronic ischemia.
引文
[1]Jin X,Kim DK,Riew TR,et al.Cellular and Subcellular Localization of Endoplasmic Reticulum Chaperone GRP78Following Transient Focal Cerebral Ischemia in Rats.Neurochem Res.2018;43(7):1348-1362.
[2]Noiri E,Nakao A,Uchida K,et al.Oxidative and nitrosative stress in acute renal ischemia.Am J Physiol Renal Physiol.2001;281(5):F948-957.
[3]Ye Z,Kong Q,Han J,et al.Circular RNAs are differentially expressed in liver ischemia/reperfusion injury model.J Cell Biochem.2018 May 18.
[4]Meloux A,Rigal E,Rochette L,et al.P1742Experimental ischemic stroke in rats induce myocardial contractile dysfunction in vivo and ex vivo and increase cardiac vulnerability to ischemia-reperfusion injury ex vivo.Eur Heart J.2017;38(suppl_1).
[5]Liu Q,Jin W N,Liu Y,et al.Brain ischemia suppresses immunity in the periphery and brain via different neurogenic innervations.Immunity.2017;46(3):474-487.
[6]Brown JM,Grosso MA,Terada LS,et al.Endotoxin pretreatment increases endogenous myocardial catalase activity and decreases ischemia-reperfusion injury of isolated rat hearts.ProcNatlAcadSci U S A.1989;86(7):2516-2520.
[7]Spreen MI,Martens JM,Knippenberg B,et al.Long‐Term Follow‐up of the PADI Trial:Percutaneous Transluminal Angioplasty Versus Drug‐Eluting Stents for Infrapopliteal Lesions in Critical Limb Ischemia.J Am Heart Assoc.2017;6(4).pii:e004877.
[8]Lee DS,Jang MJ,Cheon GJ,et al.Comparison of the cost-effectiveness of stress myocardial SPECT and stress echocardiography in suspected coronary artery disease considering the prognostic value of false-negative results.JNuclCardiol.2002;9(5):515-522.
[9]EkinciAkdemir FN,Gül?in?,Karag?z B,et al.A comparative study on the antioxidant effects of hesperidin and ellagic acid against skeletal muscle ischemia/reperfusion injury.JEnzyme Inhib Med Chem.2016;31(sup4):114-118.
[10]Chouchani ET,Pell VR,James AM,et al.A unifying mechanism for mitochondrial superoxide production during ischemia-reperfusion injury.Cell metabolism.2016;23(2):254-263.
[11]Zhang T,Zhang Y,Cui M,et al.CaMKII is a RIP3 substrate mediating ischemia-and oxidative stress-induced myocardial necroptosis.Nature medicine.2016;22(2):175.
[12]Rossi A,Wragg A,Klotz E,et al.Dynamic Computed Tomography Myocardial Perfusion ImagingClinical Perspective:Comparison of Clinical Analysis Methods for the Detection of Vessel-Specific Ischemia.CircCardiovascImaging.2017;10(4):e005505.
[13]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.
[14]杨盛家,陈兵,佟铸,等.应用激光多普勒血流仪对大鼠后肢缺血模型血流动力学的观察[J].中国普外基础与临床杂志,2011,18(2):137-141.
[15]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.
[16]黄莹,张建,陈兵,等.小鼠后肢缺血肌肉及骨髓氧分压与HIF-1α蛋白水平的关系[J].中国普外基础与临床杂志,2015,22(1):50-54.
[17]Fujiwara N,Som A T,Pham L D D,et al.A free radical scavenger edaravone suppresses systemic inflammatory responses in a rat transient focal ischemia model.Neurosci Lett.2016;633:7-13.
[18]Wyers M,Shuja F.In Patients with Acute Mesenteric Ischemia Does an Endovascular or Hybrid Approach Improve Morbidity and Mortality Compared to Open Revascularization?//Difficult Decisions in Vascular Surgery.Springer,Cham,2017:221-233.
[19]Dutkowski P,Schlegel A,Kron P,et al.Reply to“Reducing Nonanastomotic Biliary Strictures in Donation After Circulatory Death Liver Transplantation:Cold Ischemia Matters”.Ann Surg.2017;266(6):e119-e120.
[20]Manavski Y,Lucas T,Glaser S F,et al.Clonal Expansion of Endothelial Cells Contributes to Ischemia-Induced Neovascularization.Circ Res.2018;122(5):670-677.
[21]Tomita T M,Kibbe M R.Diagnostic Approach to Chronic Critical Limb Ischemia[M]//Critical Limb Ischemia.Springer,Cham.2017:137-158.
[22]Hoshijima H,Denawa Y,Mihara T,et al.Efficacy of prophylactic doses of intravenous nitroglycerin in preventing myocardial ischemia under general anesthesia:A systematic review and meta-analysis with trial sequential analysis.JClinAnesth.2017;40:16-22.
[23]Shao Q,Casin K M,Mackowski N,et al.Adenosine A1receptor activation increases myocardial protein S-nitrosothiols and elicits protection from ischemia-reperfusion injury in male and female hearts.PloS one.2017;12(5):e0177315.
[24]Spreen MI,Martens JM,Hansen BE,et al.Percutaneous transluminal angioplasty and drug-eluting stents for infrapopliteal lesions in critical limb ischemia(PADI)trial.CircCardiovascInterv.2016;9(2):e002376.
[25]Liotta F,Annunziato F,Castellani S,et al.Therapeutic Efficacy of Autologous Non-Mobilized Enriched Circulating Endothelial Progenitors in Patients With Critical Limb Ischemia-The SCELTA Trial.Circ J.2018;82(6):1688-1698.
[26]Inoue T,Abe C,Sun-sang J S,et al.Vagus nerve stimulation mediates protection from kidney ischemia-reperfusion injury throughα7nAChR+splenocytes.J Clin Invest.2016;126(5):1939-1952.
[27]Dieter RA,Kuzycz GB,Morrow DW.Iatrogenic Extremity Ischemia with the Potential for Amputation//Critical Limb Ischemia.Springer.Cham.2017:241-247.
[28]Ren C,Yao Y,Han R,et al.Cerebral ischemia induces angiogenesis in the peri-infarct regions via Notch1 signaling activation.Exp Neurol.2018;304:30-40.
[29]AbdElfattahHelal B,Elshrbiny RA,Shebl MM,et al.Study the Effect of Apocynin on Some Renal Functions in Ischemia/Reperfusion Injury in Male Albino Rats.Egyptian Journal of Hospital Medicine.2017;69(2).
[30]Sabato Sorrentino,Claudio Iaconetti,Salvatore De Rosa,et al.Hindlimb Ischemia Impairs Endothelial Recovery and Increases Neointimal Proliferation in the Carotid Artery.Scientific Reports.2018;15(8).
[31]Mantovani A,Bonapace S,Lunardi G,et al.Association between plasma ceramides and inducible myocardial ischemia in patients with established or suspected coronary artery disease undergoing myocardial perfusion scintigraphy.Metabolism.2018;85:305-312.
[32]Oerlemans M I F J,van Mil A,van Eeuwijk E,et al.Inhibition of miR-223 reduces inflammation but not adverse cardiac remodelling after myocardial ischemia-reperfusion in vivo.NCRI.2018;2(15).
[33]Zheng DY,Zhou M,Jin J,et al.Inhibition of P38 MAPKdownregulates the expression of IL-1βto protect lung from acute injury in intestinal ischemia reperfusion rats.Mediators Inflamm.2016;2016:9348037.
[34]Sunabori T,Koike M,Asari A,et al.Suppression of ischemia-induced hippocampal pyramidal neuron death by hyaluronantetrasaccharide through inhibition of Toll-like receptor 2 signaling pathway.Am J Pathol.2016;186(8):2143-2151.
[35]Nakamura K,Sano S,Fuster JJ,et al.Secreted frizzled-related protein 5 diminishes cardiac inflammation and protects the heart from ischemia-reperfusion injury.J Biol Chem.2016;291(6):2566-2575.
[2]Noiri E,Nakao A,Uchida K,et al.Oxidative and nitrosative stress in acute renal ischemia.Am J Physiol Renal Physiol.2001;281(5):F948-957.
[3]Ye Z,Kong Q,Han J,et al.Circular RNAs are differentially expressed in liver ischemia/reperfusion injury model.J Cell Biochem.2018 May 18.
[4]Meloux A,Rigal E,Rochette L,et al.P1742Experimental ischemic stroke in rats induce myocardial contractile dysfunction in vivo and ex vivo and increase cardiac vulnerability to ischemia-reperfusion injury ex vivo.Eur Heart J.2017;38(suppl_1).
[5]Liu Q,Jin W N,Liu Y,et al.Brain ischemia suppresses immunity in the periphery and brain via different neurogenic innervations.Immunity.2017;46(3):474-487.
[6]Brown JM,Grosso MA,Terada LS,et al.Endotoxin pretreatment increases endogenous myocardial catalase activity and decreases ischemia-reperfusion injury of isolated rat hearts.ProcNatlAcadSci U S A.1989;86(7):2516-2520.
[7]Spreen MI,Martens JM,Knippenberg B,et al.Long‐Term Follow‐up of the PADI Trial:Percutaneous Transluminal Angioplasty Versus Drug‐Eluting Stents for Infrapopliteal Lesions in Critical Limb Ischemia.J Am Heart Assoc.2017;6(4).pii:e004877.
[8]Lee DS,Jang MJ,Cheon GJ,et al.Comparison of the cost-effectiveness of stress myocardial SPECT and stress echocardiography in suspected coronary artery disease considering the prognostic value of false-negative results.JNuclCardiol.2002;9(5):515-522.
[9]EkinciAkdemir FN,Gül?in?,Karag?z B,et al.A comparative study on the antioxidant effects of hesperidin and ellagic acid against skeletal muscle ischemia/reperfusion injury.JEnzyme Inhib Med Chem.2016;31(sup4):114-118.
[10]Chouchani ET,Pell VR,James AM,et al.A unifying mechanism for mitochondrial superoxide production during ischemia-reperfusion injury.Cell metabolism.2016;23(2):254-263.
[11]Zhang T,Zhang Y,Cui M,et al.CaMKII is a RIP3 substrate mediating ischemia-and oxidative stress-induced myocardial necroptosis.Nature medicine.2016;22(2):175.
[12]Rossi A,Wragg A,Klotz E,et al.Dynamic Computed Tomography Myocardial Perfusion ImagingClinical Perspective:Comparison of Clinical Analysis Methods for the Detection of Vessel-Specific Ischemia.CircCardiovascImaging.2017;10(4):e005505.
[13]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.
[14]杨盛家,陈兵,佟铸,等.应用激光多普勒血流仪对大鼠后肢缺血模型血流动力学的观察[J].中国普外基础与临床杂志,2011,18(2):137-141.
[15]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.
[16]黄莹,张建,陈兵,等.小鼠后肢缺血肌肉及骨髓氧分压与HIF-1α蛋白水平的关系[J].中国普外基础与临床杂志,2015,22(1):50-54.
[17]Fujiwara N,Som A T,Pham L D D,et al.A free radical scavenger edaravone suppresses systemic inflammatory responses in a rat transient focal ischemia model.Neurosci Lett.2016;633:7-13.
[18]Wyers M,Shuja F.In Patients with Acute Mesenteric Ischemia Does an Endovascular or Hybrid Approach Improve Morbidity and Mortality Compared to Open Revascularization?//Difficult Decisions in Vascular Surgery.Springer,Cham,2017:221-233.
[19]Dutkowski P,Schlegel A,Kron P,et al.Reply to“Reducing Nonanastomotic Biliary Strictures in Donation After Circulatory Death Liver Transplantation:Cold Ischemia Matters”.Ann Surg.2017;266(6):e119-e120.
[20]Manavski Y,Lucas T,Glaser S F,et al.Clonal Expansion of Endothelial Cells Contributes to Ischemia-Induced Neovascularization.Circ Res.2018;122(5):670-677.
[21]Tomita T M,Kibbe M R.Diagnostic Approach to Chronic Critical Limb Ischemia[M]//Critical Limb Ischemia.Springer,Cham.2017:137-158.
[22]Hoshijima H,Denawa Y,Mihara T,et al.Efficacy of prophylactic doses of intravenous nitroglycerin in preventing myocardial ischemia under general anesthesia:A systematic review and meta-analysis with trial sequential analysis.JClinAnesth.2017;40:16-22.
[23]Shao Q,Casin K M,Mackowski N,et al.Adenosine A1receptor activation increases myocardial protein S-nitrosothiols and elicits protection from ischemia-reperfusion injury in male and female hearts.PloS one.2017;12(5):e0177315.
[24]Spreen MI,Martens JM,Hansen BE,et al.Percutaneous transluminal angioplasty and drug-eluting stents for infrapopliteal lesions in critical limb ischemia(PADI)trial.CircCardiovascInterv.2016;9(2):e002376.
[25]Liotta F,Annunziato F,Castellani S,et al.Therapeutic Efficacy of Autologous Non-Mobilized Enriched Circulating Endothelial Progenitors in Patients With Critical Limb Ischemia-The SCELTA Trial.Circ J.2018;82(6):1688-1698.
[26]Inoue T,Abe C,Sun-sang J S,et al.Vagus nerve stimulation mediates protection from kidney ischemia-reperfusion injury throughα7nAChR+splenocytes.J Clin Invest.2016;126(5):1939-1952.
[27]Dieter RA,Kuzycz GB,Morrow DW.Iatrogenic Extremity Ischemia with the Potential for Amputation//Critical Limb Ischemia.Springer.Cham.2017:241-247.
[28]Ren C,Yao Y,Han R,et al.Cerebral ischemia induces angiogenesis in the peri-infarct regions via Notch1 signaling activation.Exp Neurol.2018;304:30-40.
[29]AbdElfattahHelal B,Elshrbiny RA,Shebl MM,et al.Study the Effect of Apocynin on Some Renal Functions in Ischemia/Reperfusion Injury in Male Albino Rats.Egyptian Journal of Hospital Medicine.2017;69(2).
[30]Sabato Sorrentino,Claudio Iaconetti,Salvatore De Rosa,et al.Hindlimb Ischemia Impairs Endothelial Recovery and Increases Neointimal Proliferation in the Carotid Artery.Scientific Reports.2018;15(8).
[31]Mantovani A,Bonapace S,Lunardi G,et al.Association between plasma ceramides and inducible myocardial ischemia in patients with established or suspected coronary artery disease undergoing myocardial perfusion scintigraphy.Metabolism.2018;85:305-312.
[32]Oerlemans M I F J,van Mil A,van Eeuwijk E,et al.Inhibition of miR-223 reduces inflammation but not adverse cardiac remodelling after myocardial ischemia-reperfusion in vivo.NCRI.2018;2(15).
[33]Zheng DY,Zhou M,Jin J,et al.Inhibition of P38 MAPKdownregulates the expression of IL-1βto protect lung from acute injury in intestinal ischemia reperfusion rats.Mediators Inflamm.2016;2016:9348037.
[34]Sunabori T,Koike M,Asari A,et al.Suppression of ischemia-induced hippocampal pyramidal neuron death by hyaluronantetrasaccharide through inhibition of Toll-like receptor 2 signaling pathway.Am J Pathol.2016;186(8):2143-2151.
[35]Nakamura K,Sano S,Fuster JJ,et al.Secreted frizzled-related protein 5 diminishes cardiac inflammation and protects the heart from ischemia-reperfusion injury.J Biol Chem.2016;291(6):2566-2575.