迷迭香酸通过FOXO3a/VEGF信号通路对高糖环境下人视网膜微血管内皮细胞的保护作用及机制研究  

Effect of rosmarinic acid on human retinal microvascular endothelial cells in high glucose environment through FOXO3a/VEGF signaling pathway

作　　者：王化湘 刘光化 简唯求 彭辉灿 李姣 Wang Huaxiang;Liu Guanghua;Jian Weiqiu(Department of Ophthalmology and Otolaryngology,Longhui County People's Hospital,Shaoyang 422200)

机构地区：[1]隆回县人民医院眼耳鼻喉科,湖南邵阳422200 [2]南华大学附属第二医院眼科,湖南衡阳421001

出　　处：《中国现代医药杂志》2022年第4期8-13,共6页Modern Medicine Journal of China

基　　金：湖南省卫生健康委科研课题(编号:202107021669)。

摘　　要：目的探讨迷迭香酸对叉头框转录因子O3a(Forkhead box O3a,FOXO3a)/血管内皮生长因子(Vascular endothelial growth factor,VEGF)信号通路蛋白表达及对高糖环境下人视网膜微血管内皮细胞(Human retinal microvascular endothelial cells,HRMEC)的保护作用。方法将HRMEC进行高糖诱导培养,分别通过不同浓度的迷迭香酸进行处理并分组,包括空白组、低糖组、高糖组、低剂量迷迭香酸组(12.5μmol/L)、中剂量迷迭香酸组(25μmol/L)及高剂量迷迭香酸组(50μmol/L)。通过MTT检测各组HRMEC增殖活性水平,通过RT-PCR检测各组HRMEC中VEGF的mRNA相对表达水平,通过流式细胞术检测HRMEC周期分布,通过Western Blot检测各组HRMEC中VEGF、FOXO3a、增殖细胞核抗原(PCNA)、细胞周期相关蛋白CDK2、Cyclin A1的蛋白相对表达水平。结果与空白组相比,低糖环境下HRMEC各项指标比较差异无统计学意义(均P>0.05),而在高糖环境下HRMEC增殖活性显著增加,VEGF mRNA水平显著增加,细胞周期处于周期循环状态,且VEGF、FOXO3a、PCNA、CDK2、Cyclin A1各蛋白相对表达水平均明显上升(均P<0.05);与高糖组相比,经不同浓度的迷迭香酸处理后,HRMEC增殖活性显著降低,VEGF与FOXO3a mRNA水平明显降低,阻滞于G0/G1期的细胞比例明显增加,且VEGF、FOXO3a、PCNA、CDK2、Cyclin A1各蛋白相对表达水平均显著受到抑制(均P<0.05),同时其变化水平存在剂量依赖性,其中高剂量迷迭香酸组各指标变化最大。结论迷迭香酸可能通过阻滞高糖环境下HRMEC周期进程,进而抑制其细胞增殖,发挥保护作用,这一过程可能与调控FOXO3a/VEGF信号通路有关。Objective To investigate the protective effect of rosmarinic acid on FOXO3a/VEGF signaling pathway protein expression and human retinal microvascular endothelial cells(HRMEC)in high glucose environment.Methods HRMEC was natured with high glucose and treated with different concentrations of rosmarinic acid.The cells were divided into blank group,low glucose group,high glucose group,low dose rosmarinic acid group(12.5μmol/L),medium dose rosmarinic acid group(25μmol/L)and high dose romarinic acid group(50μmol/L).HRMEC proliferation activity was detected by MTT.mRNA relative expression of VEGF was detected by RT-PCR.HRMEC cycle distribution was detected by flow cytometry.The relative expression levels of VEGF,FOXO3a,PCNA,CDK2and Cyclin A1 were detected by Western Blot.Results Compared with the blank group,there was no significant difference in the expression of various indicators of HRMEC in the low glucose environment(P>0.05),while the proliferation activity of HRMEC in the high glucose environment was significantly increased and the mRNA levels of VEGF were significantly increased with the cell cycle being in a cyclic status.The relative expression levels of VEGF,FOXO3a,PCNA,CDK2 and Cyclin A1 were sharply increased(P<0.05).Comparing with the high glucose group,the proliferation activity of HRMEC,mRNA levels of VEGF and FOXO3a were decreased,and the proportion of cells blocked in G0/G1 phase was increased different concentrations of rosmarinic acid treatment.The relative expression levels of VEGF,FOXO3a,PCNA,CDK2 and Cyclin A1 were inhibited(P<0.05).The change levels were concentration-dependent,and the highest change level was found in the high dose rosmarinic acid group(P<0.05).Conclusion Rosmarinic acid may play a protective role by blocking the cycle process of HRMEC in high glucose environment and thereby inhibiting their proliferation,which may be related to the regulation of FOXO3a/VEGF signaling pathway.

关 键 词：迷迭香酸 高糖环境 叉头框转录因子O3a/血管内皮生长因子信号通路 人视网膜微血管内皮细胞 增殖 细胞周期 

分 类 号：R587.1[医药卫生—内分泌]