转录因子SP1调控MAP3K1表达影响糖尿病视网膜病变细胞凋亡和炎症反应
Transcription factor SP1 regulates expression of MAP3K1 and affects apoptosis and inflammatory response of diabetic retinopathy cells
黄丹 1邓芳祝 1谢功泽 1李巧莲2
作者信息
- 1. 郴州市第四人民医院眼科,郴州 423000
- 2. 郴州市第一人民医院北院眼科,郴州 423000
- 折叠
摘要
目的:探究转录因子SP1对糖尿病视网膜病变(DR)细胞凋亡和炎症反应的影响及分子机制.方法:构建人视网膜微血管内皮细胞(hRMECs)的DR模型,通过转染将细胞分为Control组、HG组、HG+si-NC组、HG+si-SP1组、HG+si-SP1+ oe-NC组和HG+si-SP1+oe-MAP3K1组.RT-qPCR检测SP1和MAP3K1表达;流式细胞术检测细胞凋亡;ELISA检测细胞中炎症因子TNF-α、IL-1β和IL-6含量.通过生物信息学寻找SP1与MAP3K1启动子间的结合位点,ChIP-qPCR验证SP1与MAP3K1启动子的结合情况.结果:与Control组相比,HG处理的hRMECs中SP1和MAP3K1表达均升高(P<0.05).抑制SP1表达后细胞凋亡率明显降低(P<0.05),炎症因子TNF-α、IL-1β和IL-6含量明显减少(P<0.05).抑制SP1可降低MAP3K1表达.进一步过表达MAP3K1逆转了si-SP1对HG诱导的hRMECs凋亡和炎症反应的抑制作用.结论:转录因子SP1通过促进MAP3K1表达促进DR细胞凋亡及炎症反应.
Abstract
Objective:To explore effect and molecular mechanism of transcription factor SP1 on apoptosis and inflammatory response of diabetic retinopathy(DR).Methods:To construct a DR model of human retinal microvascular endothelial cells(hRMECs),cells were divided into Control group,HG group,HG+si-NC group,HG+si-SP1 group,HG+si-SP1+oe-NC group and HG+si-SP1+oe-MAP3K1 group by transfection.RT-qPCR was used to detect expressions of SP1 and MAP3K1;flow cytometry was used to detect cell apoptosis;ELISA was used to detect contents of inflammatory factors TNF-α,IL-1β and IL-6 in cells;binding site between SP1 and MAP3K1 promoter were found by bioinformatics,and ChIP-qPCR was used to detect binding of SP1 to the MAP3K1 promoter.Results:Compared with control group,expressions of SP1 and MAP3K1 in hRMECs treated with HG were increased(P<0.05).Inhibition of SP1 expression,apoptosis rate was significantly decreased(P<0.05),and contents of inflammatory factors TNF-α,IL-1β and IL-6 were significantly decreased(P<0.05).Inhibition of SP1 reduced expression of MAP3K1.Further overexpression of MAP3K1 reversed inhibitory effect of si-SP1 on HG-induced apoptosis and inflammatory responses in hRMECs.Conclusion:Transcription factor SP1 promotes apoptosis and inflammatory response of DR cells by promoting expression of MAP3K1.
关键词
糖尿病视网膜病变/SP1/MAP3K1/凋亡/炎症反应Key words
Diabetic retinopathy/SP1/MAP3K1/Apoptosis/Inflammatory response引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据