首页> 中文期刊> 《眼科新进展》 >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

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

         

摘要

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.%目的 探讨基因沉默解离素-金属蛋白酶17(a disintegrin and metalloproteinase 17,ADAM17)对高糖环境下人视网膜微血管内皮细胞(human retinal microvascular endothelial cells,HRMECs)通透性、增殖、迁移以及蛋白表达的影响.方法 将HRMECs分为4组:正常组(5 mmol·L-1 D-葡萄糖)、高糖组(25 mmol·L-1 D-葡萄糖)、NC(siRNA阴性对照)+高糖组和si-ADAM17(ADAM17 siRNA)+高糖组.ADAM17表达采用实时定量PCR和蛋白印迹法检测;辣根过氧化酶法检测HRMECs通透性;CCK-8和BrdU检测细胞增殖;Transwell检测细胞迁移;蛋白印迹法检测p-EGFR、p-ERK和MMP9表达水平.结果 与正常组相比,高糖组ADAM17蛋白表达水平显著升高(P<0.01),NC+高糖组与高糖组相比无明显差异(P>0.05);与NC+高糖组比较,siADAM17+高糖组ADAM17蛋白表达水平显著下调(P<0.05).沉默ADAM17可降低高糖环境下HRMECs的通透性.沉默ADAM17可抑制高糖介导的细胞增殖.Transwell实验结果显示,各组相对迁移的细胞数分别为高糖组(157.00±7.93)个、NC+高糖组(169.00±10.12)个、siADAM17+高糖组(121.00±9.28)个、正常组(110.00±8.25)个.沉默ADAM17可抑制高糖介导的细胞迁移.siADAM17+高糖组较NC+高糖组p-EGFR、p-ERK和MMP9蛋白表达显著减少(均为P<0.01).结论 基因沉默ADAM17可能通过EGFR、ERK、MMP9通路降低高糖诱导的细胞通透性并抑制细胞增殖和迁移.

著录项

  • 来源
    《眼科新进展》 |2017年第3期|205-209|共5页
  • 作者单位

    710004陕西省西安市,陕西省眼科诊疗中心,西安市第四医院眼科,西安交通大学医学院附属广仁医院眼科;

    710004陕西省西安市,陕西省眼科诊疗中心,西安市第四医院眼科,西安交通大学医学院附属广仁医院眼科;

    710004陕西省西安市,陕西省眼科诊疗中心,西安市第四医院眼科,西安交通大学医学院附属广仁医院眼科;

    710004陕西省西安市,陕西省眼科诊疗中心,西安市第四医院眼科,西安交通大学医学院附属广仁医院眼科;

    710004陕西省西安市,陕西省眼科诊疗中心,西安市第四医院眼科,西安交通大学医学院附属广仁医院眼科;

    710004陕西省西安市,陕西省眼科诊疗中心,西安市第四医院眼科,西安交通大学医学院附属广仁医院眼科;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 视网膜及视神经疾病;
  • 关键词

    解离素-金属蛋白酶17; 细胞通透性; 细胞迁移;

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