摘要
目的建立和评价糖尿病下肢溃疡小鼠模型,揭示糖尿病下肢溃疡小鼠手术肢血流和病理生理的改变,初探其发病机制,为研究糖尿病外周血管病变提供基础和参考。方法小鼠分为下肢缺血组、糖尿病组和糖尿病下肢溃疡组。糖尿病下肢溃疡和糖尿病组腹腔注射链脲佐菌素(STZ)建立1型糖尿病模型。糖尿病下肢溃疡和下肢缺血组,采用高位结扎股动脉、股静脉并断离股动脉的方法建立下肢缺血模型;糖尿病组仅做假手术处理。术后第0、3、7、14、21天,用激光多普勒监测血流变化,观察肢体缺血坏死。第21天后HE切片观察组织形态变化,分析血小板-内皮细胞粘附分子-1(PECAM-1/CD31)及抗平滑肌抗体(SMA)表达。结果缺血术后,与下肢缺血组小鼠比较,糖尿病下肢溃疡组体重显著下降,肢体坏死情况更严重。术后,糖尿病下肢溃疡组和下肢缺血组小鼠手术肢血流灌注下降明显;术后第3、7、14天,糖尿病下肢溃疡组和下肢缺血组血流灌注逐渐恢复;第21天,下肢缺血组接近正常水平,而糖尿病下肢溃疡组略有下降。糖尿病组无肢体坏死情况,血流灌注无明显变化。糖尿病下肢溃疡组和下肢缺血组小鼠手术肢腓肠肌组织有肌肉结构破坏和炎症浸润,CD31表达明显增加;糖尿病下肢溃疡组和糖尿病组SMA有显著表达,而下肢缺血组表达不明显。结论成功建立了糖尿病下肢溃疡小鼠模型,与下肢缺血小鼠模型对比,该模型有明显的肢体坏死症状和血流灌注恢复障碍。该模型可用于研究糖尿病血管病变发病机制的研究以及治疗药物的筛选。
Objective To establish and evaluate a mouse model of diabetic hindlimb ulcers,study the changes in blood flow and pathophysiology in mice with diabetic hindlimb ulcers,and explore the pathogenesis of the ulcers to provide a basis and reference for studying peripheral vascular diseases in diabetes.Methods ICR mice were divided into the diabetic hindlimb ulcer,hindlimb ischemia,and diabetic groups.Mice in the diabetic hindlimb ulcer and diabetes groups were intraperitoneally injected with streptozotocin to establish a model of type 1 diabetes.Mice in the diabetic hindlimb ulcer and hindlimb ischemia groups had their femoral arteries and veins ligated and their femoral artery severed.Sham surgery was performed in the diabetic group.Blood flow changes postoperation were detected via laser Doppler,and ischemic necrosis of the limbs was observed.Twenty-one days postoperation,hematoxylin and eosin staining was used to observe morphological changes in the ischemic tissue.Changes in CD31 and SMA expressions were analyzed.Results After the operation,the weight of the diabetic hindlimb ulcer group decreased significantly compared with that of the hindlimb ischemia group,and the hindlimb necrosis was more severe.Additionally,the diabetic hindlimb ulcer and hindlimb ischemia groups showed significantly decreased blood perfusion,which recovered gradually on days 3,7,and 14 postoperation.Blood perfusion in the hindlimb ischemia group was near normal 21 days postoperation,but that of the diabetic hindlimb ulcer group decreased slightly.No significant change occurred in the diabetes group.In the diabetic hindlimb ulcer and hindlimb ischemia groups,muscle structure destruction and inflammatory infiltration occurred in the gastrocnemius tissues,and CD31 expression was increased.SMA expression was increased in the diabetic hindlimb ulcer and diabetes groups but not in the hindlimb ischemia group.Conclusions The diabetic hindlimb ulcer mouse model is successfully established.Limb necrosis and impaired recovery of blood perfusion are obvious in the diabetic hindlimb ulcer model compared with those of the hindlimb ischemia model.This model can be used to study the pathogenesis of diabetic vascular diseases and to screen therapeutic drugs.
作者
鲁珽
辛永萍
林家彬
陈民利
LU Ting;XIN Yongping;LIN Jiabin;CHEN Minli(Clinical Research Center,the Second Affiliated Hospital of Zhejiang University School of Medicine,Hangzhou 310009,China;Laboratory Animal Research Center,Zhejiang Chinese Medical University,Hangzhou 310053)
出处
《中国实验动物学报》
CAS
CSCD
北大核心
2020年第1期58-65,共8页
Acta Laboratorium Animalis Scientia Sinica
基金
浙江省科技厅公益技术应用研究(实验动物)项目(2017C37156).
关键词
糖尿病
下肢溃疡
下肢缺血
小鼠
diabetes
hindlimb ulcer
hindlimb ischemia
mouse
作者简介
鲁珽(1985—),男,实验师,硕士研究生,从事药理学研究。Email:lutinghz@163.com;通信作者:陈民利(1963—),女,教授,博士生导师,研究方向:实验动物与比较医学。Email:cmli991@zcum.edu.cn
