Effect and mechanism of miR-373-3p in diabetic retinopathy
10.11855/j.issn.0577-7402.0422.2024.0104
- VernacularTitle:miR-373-3p在糖尿病视网膜病变中的作用及其机制
- Author:
Jia JIA
1
;
Lin-Chang ZHANG
;
Hai-Xia ZHANG
Author Information
1. 天津市第五中心医院/北京大学滨海医院眼科,天津 300450
- Keywords:
miR-373-3p;
diabetic retinopathy;
vascular endothelial growth factor A;
high glucose;
retinal microvascular endothelial cells
- From:
Medical Journal of Chinese People's Liberation Army
2025;50(1):76-82
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of miR-373-3p in diabetic retinopathy(DR),as well as the underlying mechanisms.Methods Serum samples from 35 DR patients and 35 non-DR patients visiting Tianjin Fifth Central Hospital from February 2021 to February 2022 were collected,and expression levels of miR-373-3p and vascular endothelial growth factor A(VEGFA)mRNA were detected using quantitative reverse transcription polymerase chain reaction(qRT-PCR).An in vitro DR model was constructed using high glucose(HG)-treated human retinal microvascular endothelial cells(HRMEC).HRMECs were divided into control group(5 mmol/L glucose and 25 mmol/L mannitol treatment),HG group(30 mmol/L glucose treatment),HG+miR-373-3p mimic-negative control(miR-con)group(30 mmol/L glucose treatment after transfection with miR-con),HG+miR-373-3p mimic group(30 mmol/L glucose treatment after transfection with miR-373-3p),HG+miR-373-3p+vector group(30 mmol/L glucose treatment after co-transfection with miR-373-3p and vector),and HG+miR-373-3p+vascular endothelial growth factor A(VEGFA)group(30 mmol/L glucose treatment after co-transfection with miR-373-3p and VEGFA).The expression levels of miR-373-3p,VEGFA mRNA and protein were analyzed by qRT-PCR and Western blotting.CCK-8,immunofluorescence,Transwell assay,angiogenesis assay,and Western blotting were used to evaluate HRMEC proliferation,migration and angiogenesis abilities.The relationship between miR-373-3p and VEGFA was determined by dual luciferase reporter assay.Results Compared with non-DR patients,DR patients exhibited significantly lower expression levels of miR-373-3p(P＜0.05)and higher expression levels of VEGFA mRNA(P＜0.05)in serum.Compared with control group,HG group showed decreased expression of miR-373-3p(P＜0.05),increased expressions of the mRNA and protein of VEGFA(P＜0.05),higher cell viability,proliferation rate,proliferating cell nuclear antigen(PCNA)and Cylin D1 protein,and numbers of migrating cells and angiogenesis ability(P＜0.05)in HRMECs.Compared with HG+miR-con group,HG+miR-373-3p group showed increased expression of miR-373-3p(P＜0.05),decreased expressions of VEGFA(P＜0.05),lower cell viability,proliferation rate,PCNA and Cylin D1 protein(P＜0.05),and lower numbers of migrating cells and angiogenesis ability(P＜0.05)in HRMECs.Compared with HG+miR-373-3p+vector group,HG+miR-373-3p+VEGFA group showed increased expression of VEGFA(P＜0.05),higher cell viability,proliferation rate,PCNA and Cylin D1 protein,and numbers of migrating cells and angiogenesis ability(P＜0.05)in HRMECs.The results of dual luciferase reporter assay showed decreased enzymatic activity of luciferase after cotransfection of miR-373-3p and VEGFA sequence(P＜0.05).Conclusion MiR-373-3p is lowly expressed in the serum of DR patients,and its potential mechanism may involve targeting VEGFA to inhibit HG-induced HRMEC dysfunction.
