[1]孟婷宇,范晶,郑柳,等.二甲双胍对高糖诱导的RF/6A细胞增殖、迁移和血管形成的影响及其可能机制[J].眼科新进展,2023,43(5):346-351.[doi:10.13389/j.cnki.rao.2023.0070]
　MENG Tingyu,FAN Jing,ZHENG Liu,et al.Effects of metformin on proliferation and migration of choroid-retinal endothelial cells as well as angiogenesis induced by high glucose and their possible mechanisms[J].Recent Advances in Ophthalmology,2023,43(5):346-351.[doi:10.13389/j.cnki.rao.2023.0070]
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二甲双胍对高糖诱导的RF/6A细胞增殖、迁移和血管形成的影响及其可能机制/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
43卷
期数:
2023年5期
页码:
346-351
栏目:
实验研究
出版日期:
2023-05-05

文章信息/Info

Title:
Effects of metformin on proliferation and migration of choroid-retinal endothelial cells as well as angiogenesis induced by high glucose and their possible mechanisms
作者:
孟婷宇; 范晶; 郑柳; 杨彬彬; 丁芝祥
541100　广西壮族自治区桂林市，桂林医学院（孟婷宇，范晶）；541001　广西壮族自治区桂林市，桂林医学院附属医院（郑柳，杨彬彬，丁芝祥）
Author(s):
MENG Tingyu1; FAN Jing1; ZHENG Liu2; YANG Binbin2; DING Zhixiang2
1.Guilin Medical University，Guilin 541100，Guangxi Zhuang Autonomous Region，China
2.Affiliated Hospital of Guilin Medical University，Guilin 541001，Guangxi Zhuang Autonomous Region，China
关键词:
糖尿病视网膜病变; 二甲双胍; RF/6A细胞; 高糖; 细胞增殖; 细胞迁移; 血管形成; Hippo信号通路
Keywords:
diabetic retinopathy;  metformin;  choroid-retinal endothelial cells;  high glucose;  cell proliferation;  cell migration;  angiogenesis;  Hippo signaling pathway
分类号:
R774
DOI:
10.13389/j.cnki.rao.2023.0070
文献标志码:
A
摘要:
目的　研究二甲双胍（MET）对高糖诱导的恒河猴脉络膜-视网膜内皮细胞RF/6A增殖、迁移和血管形成的影响，并初步探讨其作用机制。
方法　RF/6A细胞随机分为5组：NC组、HG组、HG+15 mmol·L-1MET组、HG+30 mmol·L-1 MET组、HG+45 mmol·L-1MET组，CCK-8法检测细胞增殖情况。将RF/6A细胞随机分为4组：NC组和NC+15 mmol·L-1MET组、NC+30 mmol·L-1MET组、NC+45 mmol·L-1MET组，CCK-8法检测MET的细胞毒性。将RF/6A细胞随机分为3组：NC组、HG组和HG+15 mmol·L-1MET组，细胞划痕法和Transwell法检测细胞迁移，Matrigengel法检测细胞血管形成，RT-PCR、Western blot检测Hippo信号通路的核心成分YAP、TEAD1的mRNA及蛋白表达。
结果　CCK-8法检测结果显示，与NC组比较，HG组RF/6A 细胞的增殖活性升高（P<0.001）；与HG组比较，HG+15 mmol·L-1MET组、HG+30 mmol·L-1MET组和HG+45 mmol·L-1MET组RF/6A细胞的增殖活性均降低（均为P<0.001）。与NC组比较，NC+15 mmol·L-1MET组RF/6A细胞增殖活性无变化（P=0.227 3），因此选择15 mmol·L-1MET进行后续实验。细胞划痕法和Transwell法检测结果显示，与NC组比较，HG组RF/6A细胞的迁移率升高、迁移个数增加（均为P<0.01）；与HG组比较，HG+15 mmol·L-1MET组RF/6A细胞的迁移率降低、迁移个数减少（均为P<0.01）。Matrigengel法检测结果显示，与NC组比较，HG组RF/6A细胞的血管相对总长度增加（P<0.05）；与HG组比较，HG+15 mmol·L-1MET组RF/6A细胞的血管相对总长度减少（P<0.01）。RT-PCR检测结果显示，与NC组比较，HG组RF/6A细胞的YAP和TEAD1 mRNA相对表达量均增加（均为P<0.05）；与HG组比较，HG+15 mmol·L-1MET组RF/6A 细胞的YAP和TEAD1 mRNA相对表达量均减少（均为P<0.05）。Western blot检测结果显示，与NC组比较，HG组RF/6A 细胞的YAP和TEAD1蛋白相对表达量均增加（均为P<0.05）；与HG组比较，HG+15 mmol·L-1MET组RF/6A细胞的YAP和TEAD1蛋白相对表达量均减少（均为P<0.05）。
结论　MET可抑制高糖诱导的RF/6A细胞增殖、迁移和血管形成，下调YAP、TEAD1的mRNA及蛋白表达，推测其作用机制与Hippo信号通路有关。
Abstract:
Objective　To investigate the impact of metformin (MET) on the high glucose-induced proliferation and migration of choroid-retinal endothelial cells (RF/6A cells) and angiogenesis in rhesus monkeys and explore its mechanism preliminarily.
Methods　The RF/6A cells were randomly divided into five groups: normal control (NC) group, high glucose (HG) group, HG+15 mmol·L-1 MET group, HG+30 mmol·L-1 MET group, and HG+45 mmol·L-1 MET group, and Cell Counting Kit-8 (CCK-8) was used to detect the cell proliferation. The RF/6A cells were randomly divided into four groups: NC group, NC+15 mmol·L-1 MET group, NC+30 mmol·L-1 MET group, and NC+45 mmol·L-1 MET group, and CCK-8 was adopted to detect the cytotoxicity. The RF/6A cells were randomly divided into three groups: NC group, HG group, and HG+15 mmol·L-1 MET group, cell migration was detected by cell scratch assay and Transwell assay, angiogenesis was detected by the Matrigengel method, and the messenger ribonucleic acid (mRNA) and protein expressions of yes-associated protein (YAP) and transcriptional enhanced associate domain 1 (TEAD1), core components of the Hippo signaling pathway, were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.
Results　The CCK-8 results showed that the proliferative activity of RF/6A cells in the HG group was higher than that in the NC group (P<0.001), while that in the HG+15 mmol·L-1 MET group, HG+30 mmol·L-1 MET group and HG+45 mmol·L-1 MET group was lower than the HG group (all P<0.001). As the proliferative activity of RF/6A cells in the NC+15 mmol·L-1 MET group had no difference from that in the NC group (P=0.227 3), the concentration of 15 mmol·L-1 was selected for the follow-up experiment. The results of cell scratch assay and Transwell assay showed that the migration rate and migration number of RF/6A cells in the HG group were higher than those in the NC group (both P<0.01), while those in the HG+15 mmol·L-1 MET group were lower than the HG group (both P<0.01). The detection using Matrigengel method showed that the total vessel length of RF/6A cells in the HG group was higher than that in the NC group (P<0.05), and that in the HG+15 mmol·L-1 MET group was lower than the HG group (P<0.01). The RT-PCR results showed that the mRNA expressions of YAP and TEAD1 in RF/6A cells in the HG group were higher than those in the NC group (both P<0.05), while those in the HG+15 mmol·L-1 MET group were lower than the HG group (both P<0.05). The Western blot results showed that the relative protein expressions of YAP and TEAD1 in RF/6A cells in the HG group were higher than those in the NC group (both P<0.05), while those in the HG+15 mmol·L-1 MET group were lower than the HG group (both P<0.05).
Conclusion　MET can inhibit the proliferation and migration of RF/6A cells as well as angiogenesis induced by high glucose, and down-regulate the mRNA and protein expressions of YAP and TEAD1. It is speculated that its mechanism is related to the Hippo signaling pathway.

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备注/Memo

备注/Memo:
国家自然科学基金项目（编号：82160197）
更新日期/Last Update: 2023-05-05