[关键词]
[摘要]
目的：探讨达格列净对高糖诱导人视网膜血管内皮细胞(HRVECs)凋亡、氧化应激的影响及其对FOXO4的调控作用。
方法：采用高糖诱导HRVECs建立细胞损伤模型(高糖组)，实验分组：高糖+达格列净低剂量组(1ng/L)、高糖+达格列净中剂量组(5ng/L)、高糖+达格列净高剂量组(10ng/L)、高糖+达格列净高剂量+pcDNA组、高糖+达格列净高剂量+pcDNA-FOXO4组、正常糖组(5.5mmol/L D-葡萄糖)、高糖+si-NC组、高糖+si-FOXO4组； 采用流式细胞术检测细胞凋亡率； 根据试剂盒检测SOD、MDA的水平； 采用Western blot法检测FOXO4蛋白表达量。
结果：与正常糖组比较，高糖组细胞凋亡率升高，MDA的水平升高，FOXO4蛋白水平升高，SOD的水平降低(均P<0.05)； 与高糖组比较，高糖+达格列净中剂量组、达格列净高剂量组细胞凋亡率降低，MDA的水平降低，FOXO4蛋白水平降低，SOD的水平升高(均P<0.05)； 与高糖+达格列净高剂量+pcDNA组比较，高糖+达格列净高剂量+pcDNA-FOXO4组细胞凋亡率升高，MDA的水平升高，SOD的水平降低(均P<0.05)。
结论：达格列净可通过下调FOXO4而抑制高糖诱导的HRVECs氧化应激及抑制细胞凋亡从而减轻细胞损伤。
[Key word]
[Abstract]
AIM: To explore the effect of dapagliflozin on the apoptosis and oxidative stress of high glucose-induced human retinal vascular endothelial cells and its regulatory effect on forkhead FOXO4.
METHODS: High glucose-induced human retinal vascular endothelial cells(HRVECs)were used to establish a cell injury model(high glucose group). Experimental groups include high glucose+dapagliflozin low-dose group(1ng/L dapagliflozin), high glucose+dapagliflozin medium-dose group(5ng/L dapagliflozin), high glucose+dapagliflozin high-dose group(10ng/L dapagliflozin), high glucose+dapagliflozin high-dose+pcDNA group, high glucose+dapagliflozin high-dose+pcDNA-FOXO4 group, and normal sugar group(5.5mmol/L D-glucose). Flow cytometry was used to detect the apoptosis rate. The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)were tested with corresponding kits. Western blot assay was used to detect the protein level of FOXO4.
RESULTS: Compared with the normal sugar group, the apoptosis rate(P<0.05), the level of MDA(P<0.05)and FOXO4(P<0.05)were increased, but the level of SOD was decreased(P<0.05)in high-glucose group. Compared with the high glucose group, cell apoptosis rate(P<0.05), the level of MDA(P<0.05)and the protein level of FOXO4 were decreased(P<0.05), but the level of SOD was increased(P<0.05)in high glucose+medium-dose dapagliflozin group and high glucose+high-dose dapagliflozin group. Compared with high glucose+dapagliflozin high-dose+pcDNA group, the apoptosis rate(P<0.05)and the level of MDA(P<0.05)were increased, but the level of SOD was decreased(P<0.05)in high glucose+dapagliflozin high-dose+pcDNA-FOXO4 group(P<0.05).
CONCLUSION: Dapagliflozin could inhibit oxidative stress and cell apoptosis in high glucose-induced HRVECs by down-regulating FOXO4, thereby reducing cell damage.
[中图分类号]
[基金项目]
