Effects of ADAM17 on high-glucose mediated permeability, proliferation, migration and proteins expression in human retinal microvascular endothelial cells
10.13389/j.cnki.rao.2017.0053
- VernacularTitle:基因沉默解离素-金属蛋白酶17(ADAM17)对高糖环境下人视网膜微血管内皮细胞通透性、增殖、迁移以及蛋白表达的影响
- Author:
Bochang LV
;
Bei LIU
;
Jing LIU
;
Xinguang YANG
;
Xiquan ZHAO
;
Zhongqiao ZHU
- Keywords:
a disintegrin and metalloproteinase 17 (ADAM17);
cell permeability;
cell migration
- From:
Recent Advances in Ophthalmology
2017;37(3):205-209
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects and mechanisms of a disintegrin and metalloproteinase 17 (ADAM17) on high-glucose mediated permeability,proliferation and migration in human retinal microvascular endothelial cells (HRMECs).Methods HRMECs were divided into 4 groups:normal group (5 mmol · L-1 glucose),high glucose group (25 mmol · L-1 glucose),NC (Negative control for siRNA) + high glucose group and siADAM17 (ADAM17 siRNA) + high glucose group.The expression of ADAM17 was detected using real time PCR and Western blot.Horseradish Peroxidase (HRP) was used to detect the permeability of HRMECs.Cell Counting Kit-8 (CCK-8)and BrdU were used to evaluate cell proliferation.Cell migration was determined using Transwell assay.In addition,the expression of p-EGFR,p-ERK and MMP9 was assayed using Western blot.Results Compared with normal group,the mRNA and protein levels of ADAM17 were increased in high glucose group (P ＜ 0.01).ADAM17 expression of siADAM17 + high glucose group was markedly reduced compared with NC + high glucose group.High glucose increased the permeability of HRP comparison to normal group,whereas in siADAM17 + high glucose group the permeability of HRP was reduced compared with NC + high glucose group.The optical density of HRMECs was decreased in siADAM17 + high glucose group 1.53 ± 0.29 in comparison with NC + high glucose group 2.43 ± 0.25,as well as the content of BrdU-incorporation(P ＜ 0.05).The number of migrated cells in high glucose group,NC + high glucose group,siADAM17 + high glucose group and normal group were 157.00 ± 7.93,169.00 ± 10.12,121.00 ± 9.28,110.00 ±8.25,respectively.Moreover,the expression of p-EGFR,p-ERK and MMP9 in siADAM17 +high glucose group was decreased compared with NC + high glucose group (all P ＜0.01).Conclusion SiADAM17 can reduce the cell permeability,suppressed and migration induced by high glucose via EGFR/ERK/MMP9 signaling pathway.
