大黄素调节HIF-1α/VEGF通路对糖尿病大血管病变大鼠血管内皮细胞损伤的影响

Effects of emodin modulation of the HIF-1α/VEGF pathway on vascular endothelial cells damage in diabetic macroangiopathy rats

目的探讨大黄素(EM)调节缺氧诱导因子1α(HIF-1α)/血管内皮生长因子(VEGF)信号通路对糖尿病大血管病变大鼠血管内皮细胞损伤的影响。方法将SD大鼠分为空白组和造模组,造模组用高脂高糖联合N-硝基-L-精氨酸甲酯喂养大鼠以构建糖尿病大血管病变模型,空白组给予普通饲料饲养。从空白组、造模组大鼠胸主动脉中成功分离出来的血管内皮细胞分别命名为对照组和造模组。造模组的血管内皮细胞分为模型组、二甲基草酰甘氨酸(DMOG)组(10μmol·L^(-1) DMOG)、联合组(80 mg·L^(-1) EM+10μmol·L^(-1) DMOG)和低、中、高剂量实验组(20、40、80 mg·L^(-1) EM)。用流式细胞术检测大鼠血管内皮细胞的凋亡情况,用蛋白质印迹法检测大鼠血管内皮细胞中HIF-1α和VEGF蛋白的表达水平。结果中、高剂量实验组和DMOG组、联合组、模型组、对照组的血管内皮细胞凋亡率分别为(10.18±0.36)%、(6.28±0.20)%、(24.96±1.18)%、(12.36±0.49)%、(18.76±0.68)%和(4.59±0.26)%,HIF-1α蛋白相对表达水平分别为0.96±0.07、0.78±0.06、2.03±0.12、1.05±0.13、1.58±0.12和0.69±0.05,VEGF蛋白相对表达水平分别为0.59±0.05、0.23±0.02、0.98±0.06、0.63±0.04、0.86±0.07和0.11±0.01。模型组的上述指标与对照组、DMOG组上述指标和中、高剂量实验组比较,联合组的上述指标与高剂量实验组比较,在统计学上差异均有统计学意义(均P<0.05)。结论EM可能通过抑制HIF-1α/VEGF通路,从而改善糖尿病大血管病变大鼠血管内皮细胞损伤。

Objective To investigate the impact of emodin(EM)on vascular endothelial cell injury in rats with diabetes macroangiopathy by regulating hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathway.Methods SD rats were divided into blank group and modeling group,the rats in the modeling group were fed with high fat and high sugar combined with N-nitro-L-arginine methyl ester to build the diabetes macroangiopathy model,and the blank group was fed with ordinary diet.The vascular endothelial cells successfully isolated from the thoracic aorta of rats in blank group and modeling group were named control group and model group,respectively.The vascular endothelial cells in the modeling group were divided into model group,dimethyloxallyl glycine(DMOG)group(10μmol·L^(-1) DMOG),combined group(80 mg·L^(-1) EM+10μmol·L^(-1) DMOG)and experimental-L,-M,-H groups(20,40,80 mg·L^(-1) EM).The apoptosis of rat vascular endothelial cells was detected by flow cytometry;Western blot was applied to detect the expression of HIF-1αand VEGF proteins in rat vascular endothelial cells.Results The apoptosis rates of vascular endothelial cells in experimental-M,-H groups,DMOG group,combined group,model group and control group were(10.18±0.36)%,(6.28±0.20)%,(24.96±1.18)%,(12.36±0.49)%,(18.76±0.68)% and(4.59±0.26)%;HIF-1α protein levels were 0.96±0.07,0.78±0.06,2.03±0.12,1.05±0.13,1.58±0.12 and 0.69±0.05;VEGF protein levels were 0.59±0.05,0.23±0.02,0.98±0.06,0.63±0.04,0.86±0.07 and 0.11±0.01.The above indexes in the model group were compared with the control,DMOG,experimental-M and experimental-H groups,and the above indexes in the combined group were compared with the experimental-H group,and the differences were statistically significant(all P<0.05).Conclusion EM may inhibit HIF-1α/VEGF pathway to improve vascular endothelial cell injury in rats with diabetes macroangiopathy.