PMID- 34926194 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20231108 IS - 2222-3959 (Print) IS - 2227-4898 (Electronic) IS - 2222-3959 (Linking) VI - 14 IP - 12 DP - 2021 TI - Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose. PG - 1820-1827 LID - 10.18240/ijo.2021.12.03 [doi] AB - AIM: To determine the effect of exosomes derived from human umbilical cord blood mesenchymal stem cells (hUCMSCs) on the expression of vascular endothelial growth factor A (VEGF-A) in human retinal vascular endothelial cells (HRECs). METHODS: Exosomes were isolated from hUCMSCs using cryogenic ultracentrifugation and characterized by transmission electron microscopy, Western blotting and nanoparticle tracking analysis. HRECs were randomly divided into a normal control group (group A), a high glucose model group (group B), a high glucose group with 25 microg/mL (group C), 50 microg/mL (group D), and 100 microg/mL exosomes (group E). Twenty-four hours after coculture, the cell proliferation rate was detected using flow cytometry, and the VEGF-A level was detected using immunofluorescence. After coculture 8, 16, and 24h, the expression levels of VEGF-A in each group were detected using PCR and Western blots. RESULTS: The characteristic morphology (membrane structured vesicles) and size (diameter between 50 and 200 nm) were observed under transmission electron microscopy. The average diameter of 122.7 nm was discovered by nanoparticle tracking analysis (NTA). The exosomal markers CD9, CD63, and HSP70 were strongly detected. The proliferation rate of the cells in group B increased after 24h of coculture. Immunofluorescence analyses revealed that the upregulation of VEGF-A expression in HRECs stimulated by high glucose could be downregulated by cocultured hUCMSC-derived exosomes (F=39.03, P<0.01). The upregulation of VEGF-A protein (group C: F=7.96; group D: F=17.29; group E: F=11.89; 8h: F=9.45; 16h: F=12.86; 24h: F=42.28, P<0.05) and mRNA (group C: F=4.137; group D: F=13.64; group E: F=22.19; 8h: F=7.253; 16h: F=16.98; 24h: F=22.62, P<0.05) in HRECs stimulated by high glucose was downregulated by cocultured hUCMSC-derived exosomes (P<0.05). CONCLUSION: hUCMSC-derived exosomes downregulate VEGF-A expression in HRECs stimulated by high glucose in time and concentration dependent manner. CI - International Journal of Ophthalmology Press. FAU - He, Guang-Hui AU - He GH AD - Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300070, China. AD - Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, China. AD - Ophthalmic Center of Xinjiang Production and Construction Corps Hospital, Urumqi 830002, Xinjiang Uygur Autonomous Region, China. FAU - Ma, Ying-Xue AU - Ma YX AD - Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300070, China. AD - Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, China. FAU - Dong, Meng AU - Dong M AD - Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300070, China. AD - Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, China. FAU - Chen, Song AU - Chen S AD - Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300070, China. AD - Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, China. FAU - Wang, Yu-Chuan AU - Wang YC AD - Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300070, China. AD - Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, China. FAU - Gao, Xiang AU - Gao X AD - Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, China. AD - Medical College of NanKai University, Tianjin 300000, China. FAU - Wu, Bin AU - Wu B AD - Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300070, China. AD - Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, China. FAU - Wang, Jian AU - Wang J AD - Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300070, China. AD - Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, China. FAU - Wang, Jun-Hua AU - Wang JH AD - Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300070, China. AD - Tianjin Eye Hospital, Tianjin Eye Institute, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, China. LA - eng PT - Journal Article DEP - 20211218 PL - China TA - Int J Ophthalmol JT - International journal of ophthalmology JID - 101553860 PMC - PMC8640780 OTO - NOTNLM OT - coculture OT - exosomes OT - mesenchymal stem cells OT - retinal vascular endothelial cells OT - vascular endothelial growth factor A EDAT- 2021/12/21 06:00 MHDA- 2021/12/21 06:01 PMCR- 2021/12/18 CRDT- 2021/12/20 06:16 PHST- 2021/08/13 00:00 [received] PHST- 2021/10/22 00:00 [accepted] PHST- 2021/12/20 06:16 [entrez] PHST- 2021/12/21 06:00 [pubmed] PHST- 2021/12/21 06:01 [medline] PHST- 2021/12/18 00:00 [pmc-release] AID - ijo-14-12-1820 [pii] AID - 10.18240/ijo.2021.12.03 [doi] PST - epublish SO - Int J Ophthalmol. 2021 Dec 18;14(12):1820-1827. doi: 10.18240/ijo.2021.12.03. eCollection 2021.