Small interfering RNA targeting PGC-1α inhibits VEGF expression and tube formation in human retinal vascular endothelial cells

• Published in: International journal of ophthalmology 2015-10, Vol.8 (5), p.877-883
• Main Authors: Jiang, Jian, Zhang, Lu, Xia, Xiao-Bo
• Format: Article
• Language: English
• Subjects: Basic Research; peroxisome proliferator-activated receptor-γ coactivator-1α; retinal vascular endothelial cell; small interfering RNA; tube formation; vascular endothelial growth factor
• ISSN: 2222-3959; 2227-4898
• DOI: 10.3980/j.issn.2222-3959.2015.05.05

AIM: To determine whether small interfering RNA(si RNA)of PGC-1α could inhibit vascular endothelial growth factor(VEGF) expression and tube formation in human retinal vascular endothelial cells(h RVECs).·METHODS: h RVECs transfected with peroxisome proliferator-activated receptor-γ coactivator-1α(PGC-1α)si RNA were incubated for 24 h and then placed into a normoxic(20%, O2) or hypoxic(1%, O2) environment for another 16 h. PGC-1α m RNA and protein levels were detected by real-time PCR and Western blot. VEGF m RNA and protein levels were detected by real-time PCR and ELISA. Cell proliferation was evaluated by Brd U incorporation assay. Forty-eight hours after si RNA transfection, h RVECs were planted into Matrigel-coated plates and cultured under normoxic(20%, O2) or hypoxic(1%, O2) conditions for another 48 h. The tube formation of h RVECs was observed under an optical microscope and quantified by counting the number of branch points and calculating the total tube length.·RESULTS: PGC-1α m RNA and protein levels were significantly reduced by PGC-1α si RNA, and VEGF m RNA and protein levels also decreased significantly. The percentage of Brd U-labeled cells in si PGC-1α groups were significantly decreased compared with control si RNA groups under normoxia and hypoxia in cell proliferation assay. In the tube formation assay, PGC-1αsi RNA treated cells formed significantly fewer tubes.·CONCLUSION: Blocking PGC-1α expression can inhibit VEGF expression in h RVECs and inhibit their ability to form tubes under both normoxic and hypoxic conditions.