糖尿病后肢缺血大鼠模型的建立与评估
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国家自然科学基金项目(编号:81470587)。


Establishment and evaluation of a diabetic rat model of hindlimb ischemia
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    摘要:

    目的研究糖尿病肢体缺血模型的建立方法,为糖尿病下肢缺血和糖尿病足研究提供试验方法。方法取25只Wistar大鼠分为模型组、对照组及假手术组,模型组腹腔注射链脲佐菌素(60 mg/kg)诱发糖尿病,血糖达16.8 mmol/L模型成功。糖尿病造模成功后,手术结扎大鼠左侧股动脉、剪断周围动脉分支,建立左后肢缺血模型。术后激光多普勒监测动态监测双后肢血流,同时观察血糖、体重及尿量变化。术后21 d取后肢腓肠肌行肌肉HE染色,取结扎处股动脉免疫荧光染色观察血管平滑肌增殖情况,CD31免疫组织化学染色检测后肢肌肉毛细血管密度。结果在治疗后21 d时,无论是最大膀胱容量、漏尿点压力还是收缩力/肌重比,IGF-1组、电刺激组治疗效果更佳;而IGF-1组与电刺激组两组之间比较差异无显著性(P>0.05)。结论成功建立有效、简便易行的糖尿病后肢缺血模型,可用于糖尿病肢体缺血及糖尿病足的药物干预研究。

    Abstract:

    Objective To establish a diabetic rat model of hindlimb ischemia, and provide a test method for diabetic limb ischemia and diabetic foot study. Methods Twenty-five Wistar rats were divided into three groups:the model group and control group (n=10), and sham operated group (n=5). The rats of model group were induced by intraperitoneal injection of streptozotocin (STZ, 60 mg/kg), with the blood glucose level over 16.8 mmol/L, while the rats of control group were injected with normal saline. All the rats had the left femoral artery and its branches ligated, and then Doppler scan blood flow analysis was performed for the two hind limbs after operation at different time points. In the meantime, the changes of body weight and blood glucose were observed. The animals were sacrificed at 21days after operation. HE staining was used to observe the pathological changes of gastrocnemius and femoral arteries. Capillary density and intima hyperplasia were examined using immunostaining for CD31 and a-SMA. Results Blood glucose of the model group rats was significantly increased as well as the quantity of urine and feces,and associated with weight loss, which were maintained for more than 21 days. The blood flow of control rats was markedly decreased immediatly after operation, reached to the lowest point after surgery, and recovered from 7-14 days after operation. In contrast to the control group, the model group showed a marked reduction in blood flow in the ischemic hind limb. Pathological examination revealed remarkable vascular atrophy and a significantly reduced number of vessels per high power field in the gastrocnemius muscle of model group rats with respect to the control group, and the femoral arteries of model rats were more narrowed than that of the control rats. Conclusions In this study, an effective, convenient diabetic rat model of hindlimb ischemia is successfully established which will facilitate the studies of drug intervention for diabetic limb ischemia and diabetic foot in the future.

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齐一侠,张超,崔世军,罗涛,谷涌泉,李建新,张建,汪忠镐.糖尿病后肢缺血大鼠模型的建立与评估[J].中国实验动物学报,2015,23(6):586~590,596.

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  • 收稿日期:2015-07-12
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  • 在线发布日期: 2015-12-30
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