摘要
目的探讨松果菊苷(ECH)对高糖诱导的人视网膜毛细血管内皮细胞(HRCECs)增殖、迁移及新生血管形成的影响及其作用机制。方法将HRCECs分为5组正常对照(Con)组、高糖组(HG,25 mmol·L^(-1) D-葡萄糖)、HG+1μmol·L^(-1) ECH组、HG+5μmol·L^(-1) ECH组和HG+25μmol·L^(-1) ECH组。利用细胞管腔形成实验检测各组HRCECs血管新生能力,用MTT法检测各组HRCECs的细胞活性;用细胞集落形成实验检测各组HRCECs集落形成能力;用Transwell法检测各组HRCECs的迁移能力;用Ki67抗原(Ki67)免疫荧光染色评估各组HRCECs增殖情况;用免疫荧光染色观察各组HRCECs中细胞间黏附分子-1(ICAM-1)阳性表达;采用Western blot法检测各组HRCECs中缺氧诱导因子-1α(HIF-1α)蛋白和血管内皮生长因子(VEGF)蛋白的表达。结果与Con组比较,HG组HRCECs的血管新生能力、细胞活性、集落形成能力、Ki67阳性细胞比例、迁移能力以及细胞中ICAM-1、HIF-1α蛋白和VEGF蛋白表达均明显升高(均为P<0.001);与HG组比较,HG+1μmol·L^(-1) ECH组、HG+5μmol·L^(-1) ECH组和HG+25μmol·L^(-1) ECH组HRCECs的上述指标随ECH剂量的增加而递减(P<0.05、P<0.01、P<0.001)。结论ECH具有抑制HG诱导的HRCECs增殖、迁移及血管新生的作用,这些作用可能与其抑制HIF-1α/VEGF信号通路有关。
Objective To investigate the effect of echinacoside(ECH)on proliferation,migration and angiogenesis of human retinal capillary endothelial cells(HRCECs)induced by high glucose,and to analyze its related mechanism.Methods HRCECs were divided into five groups normal control group(Con),high glucose group(HG,25 mmol·L^(-1) D-glucose),HG+1μmol·L^(-1) ECH group,HG+5μmol·L^(-1) ECH group,and HG+25μmol·L^(-1) ECH group.Angiogenesis of HRCECs in each group was measured by tube formation assay;viability of HRCECs in each group was measured by MTT assay;colony formation of HRCECs in each group was tested by colony formation assay;migration of HRCECs in each group was detected by Transwell assay;proliferation of HRCECs in each group was evaluated by Ki67 antigen(Ki67)immunofluorescence staining;the positive expression of intercellular adhesion molecule 1(ICAM-1)of HRCECs in each group was observed by immunofluorescence staining;and the expression levels of hypoxia-inducible factor 1α(HIF-1α)and vascular endothelial growth factor(VEGF)in HRCECs in each group were detected by Western blot.Results Compared with the Con group,the angiogenesis,viability,colony formation,proportion of Ki67-positive cells,and migration of HRCECs,as well as the expression levels of ICAM-1,HIF-1αand VEGF in the HG group were significantly increased(all P<0.001).Compared with the HG group,the above indexes in the HG+1μmol·L^(-1) ECH group,HG+5μmol·L^(-1) ECH group and HG+25μmol·L^(-1) ECH group were decreased with the increase of ECH dose(P<0.05,P<0.01,and P<0.001).Conclusion ECH can inhibit the proliferation,migration and angiogenesis of HRCECs induced by HG,which may be related to its inhibition on HIF-1α/VEGF signaling pathway.
作者
毛羽佳
殷俏
郑琳璐
曾军
MAO Yujia;YIN Qiao;ZHENG Linlu;ZENG Jun(Department of Ophthalmology,the First Affiliated Hospital of Jiangxi Medical College,Shangrao 334000,Jiangxi Province,China;Department of Ophthalmology,the Second Xiangya Hospital of Central South University,Changsha 410012,Hunan Province,China)
出处
《眼科新进展》
CAS
北大核心
2022年第4期272-277,共6页
Recent Advances in Ophthalmology
基金
江西省卫健委基金项目(编号202141027)。
关键词
松果菊苷
糖尿病视网膜病变
视网膜毛细血管内皮细胞
增殖
迁移
血管新生
HIF-1α/VEGF信号通路
echinacoside
diabetic retinopathy
retinal capillary endothelial cells
proliferation
migration
angiogenesis
hypoxia-inducible factor 1α/vascular endothelial growth factor signaling pathway
