糖尿病基础研究

　　【摘要】 目的 测定血管抑素结合蛋白(Amot)mRNA及蛋白在糖尿病后肢缺血性溃疡小鼠创面肉芽组织中的表达，探讨高糖及缺血对其表达的影响。 方法 建立糖尿病后肢缺血性溃疡小鼠模型并根据有无糖尿病及缺血进行分组。RT-PCR测序鉴定Amot mRNA表达。实时荧光定量PCR检测各组Amot mRNA表达差异，Western印迹方法测定各组Amot蛋白表达。结果 各组后肢溃疡肉芽组织中均有Amot mRNA表达。在糖尿病缺血性溃疡组表达量最少(0.0014±0.0005)，与其他组比较差异均有统计学意义(P<0.01)。非糖尿病非缺血性溃疡组表达量最多，与糖尿病溃疡两组比较[(0.0187±0.0119) vs (0.0014±0.0005)，(0.0187±0.0119)vs (0.0091±0.0049)]差异有统计学意义(P<0.01)。p130-Amot在糖尿病缺血性溃疡组表达量(0.37±0.01)最少，非糖尿病非缺血性溃疡组表达量(0.88±0.02)最多，与其他组比较差异均有统计学意义(P<0.01)。p80-Amot在各组间表达比较差异无统计学意义。 结论 Amot参与了糖尿病溃疡愈合过程。高糖和缺血可造成Amot表达下降，高糖的影响更显著。

　　【关键词】血管抑素结合蛋白;糖尿病;缺血性溃疡

　　【Abstract】 Objective Angiomotin (Amot) mRNA and protein expressions in the wound granulation tissues of the diabetic mouse model with an ischemic ulcer at its hindlimb were assayed. And we investigated high glucose and ischemia effect on Amot expression. Methods A diabetic mouse model with an ischemic ulcer at its hindlimb was established. And 24 ICR mice were divided into four groups according to blood glucose and ischemic or not. The production of RT-PCR was sequenced and Amot mRNA expression was identified. Real-time fluorescent quantitation PCR was used to determine the variance of the Amot mRNA expression in different groups. Western blot was used to determine the Amot expression. Results Amot mRNA was expressed in the wound granulation tissues of these four groups. The Amot mRNA expressions in the wound granulation tissues of mouse hindlimb ulcers were different. Compared with other groups, the diabetic ischemic ulcer group had the lowest level(0.0014±0.0005)(P<0.01). The non-diabetic non-ischemic ulcer group had higher level of expression than that of the two diabetic ulcer groups[(0.0187±0.0119)vs(0.0014±0.0005), (0.0187±0.0119)vs(0.0091±0.0049)，P<0.01]. In these 4 groups mice, expression of p130-Amot was the lowest in the diabetic ischemic ulcer group(0.37±0.01)and was the highest in the non-diabetic non-ischemic ulcer group(0.88±0.02) (P<0.01). There was no significant different in expression of p80-Amot between groups. Conclusion Amot participates the process of the wound-healing in diabetic ulcers. High glucose and ischemia decreases the Amot expression in the wound granulation tissues, and high glucose has a more significant negtive effect.

　　【Key words】 Angiomotin (Amot);Diabetes;Ischemic ulcer