糖尿病大鼠后肢缺血与内皮型一氧化氮合酶解耦联的关系

目的 探讨糖尿病大鼠后肢缺血损伤的血管内皮功能障碍与内皮型一氧化氮合酶（endothelial nitric oxide synthase,eNOS）解耦联的关系.方法 15只SD大鼠随机分为正常组（5只）、糖尿病组（5只）及糖尿病后肢缺血组（5只）,建立糖尿病模型及糖尿病后肢缺血模型.建模2周后,用酶联免疫法检测各组股动脉一氧化氮（NO）的表达;用二氢乙锭（dihydroethidium,DHE）荧光探针法检测各组股动脉冰冻切片中超氧阴离子（O2-）的表达;Western blot检测各组股动脉1型三磷酸鸟苷环化水解酶（GPT cy-clohydrolase 1,GTPCH-1）的表达.结果 ①与正常组比较,糖尿病组股动脉NO表达降低,糖尿病后肢缺血组降低更为显著,3组间比较差异具有统计学意义（P=0.000,P=0.000,P=0.001）.②与正常组比较,糖尿病组O2-的表达明显增加,但不及糖尿病后肢缺血组.③与正常组比较,糖尿病组股动脉GTPCH-1的表达降低,糖尿病后肢缺血组降低更为显著,3组间比较差异具有统计学意义（P =0.001,P=0.000,P=0.012）.结论 在糖尿病大鼠后肢缺血血管损伤中存在eNOS解耦联.

The relationship between hind limb ischemia of diabetic rats and endothelial nitric oxide synthase uncoupling

Objective To investigate the relationship between vascular injury of diabetic rats with hind limb ischemia and endothelial nitric oxide synthase（eNOS） uncoupling.Methods 15 SD rats were randomly divided into 3 groups：control group （n =5）,diabetes group （n =5） and diabetes ischemia group （n =5）.Two weeks after the diabetic rat model and hind limb ischemia of diabetic rat model were set up,Enzyme linked immunosorbent assay（ELISA） was used to detect nitric oxide（NO） expression in the rat femoral artery; Dihydroethidium（DHE） fluorescent probe was used to detect superoxide（O2-） production in the frozen section of femoral artery; and Western blot was used to examine GTP cyclohydrolase 1 （GTPCH-1）expression in femoral artery.Results ①Compared with the control group,NO expression of femoral artery in diabetes group was significantly reduced,diabetes ischemia group decreased more significantly,and the difference between the 3 groups had statistical significance（P =0.000,P =0.000,P =0.001）.②Frozen section showed：compared with the control group,O2-expression of diabetes group significantly increased,but not as much as diabetes hind limb ischemia group.③Western blot results showed that compared with the control group,the expression of GTPCH-1 in diabetes group reduced,diabetes hind limb ischemia group decreased more significantly,and the difference between the 3 groups had statistical significance （P =0.001,P =0.000,P =0.012）.Conclusion eNOS uncoupling is present in diabetic rats with hind limb ischemia and vascular injury.