Abstract
Specnuezhenide (SPN), one of the main ingredients of Chinese medicine "Nü-zhen-zi", has anti-angiogenic and vision improvement effects. However, studies of its effect on retinal neovascularization are limited so far. In the present study, we established a vascular endothelial growth factor A (VEGFA) secretion model of human acute retinal pigment epithelial-19 (ARPE-19) cells by exposure of 150 μM CoCl₂ to the cells and determined the VEGFA concentrations, the mRNA expressions of VEGFA, hypoxia inducible factor-1α (HIF-1α) & prolyl hydroxylases 2 (PHD-2), and the protein expressions of HIF-1α and PHD-2 after treatment of 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1, 1.0 μg/mL) or SPN (0.2, 1.0 and 5.0 μg/mL). Furthermore, rat pups with retinopathy were treated with SPN (5.0 and 10.0 mg/kg) in an 80% oxygen atmosphere and the retinal avascular areas were assessed through visualization using infusion of ADPase and H&E stains. The results showed that SPN inhibited VEGFA secretion by ARPE-19 cells under hypoxia condition, down-regulated the mRNA expressions of VEGFA and PHD-2 slightly, and the protein expressions of VEGFA, HIF-1α and PHD-2 significantly in vitro. SPN also prevented hypoxia-induced retinal neovascularization in a rat model of oxygen-induced retinopathy in vivo. These results indicate that SPN ameliorates retinal neovascularization through inhibition of HIF-1α/VEGF signaling pathway. Therefore, SPN has the potential to be developed as an agent for the prevention and treatment of diabetic retinopathy.
Keywords:
angiogenesis; hypoxia-inducible factor-1; oxygen-induced retinopathy; specnuezhenide; vascular endothelial growth factor.
MeSH terms
-
Angiogenesis Inhibitors / isolation & purification
-
Angiogenesis Inhibitors / pharmacology*
-
Animals
-
Cell Hypoxia
-
Cell Line
-
Cobalt / pharmacology
-
Diabetic Retinopathy / drug therapy
-
Diabetic Retinopathy / genetics
-
Diabetic Retinopathy / metabolism
-
Diabetic Retinopathy / pathology
-
Disease Models, Animal
-
Epithelial Cells / cytology
-
Epithelial Cells / drug effects
-
Epithelial Cells / metabolism
-
Gene Expression Regulation
-
Glucosides / isolation & purification
-
Glucosides / pharmacology*
-
Humans
-
Hypoxia / complications
-
Hypoxia / drug therapy*
-
Hypoxia / genetics
-
Hypoxia / pathology
-
Hypoxia-Inducible Factor 1, alpha Subunit / antagonists & inhibitors*
-
Hypoxia-Inducible Factor 1, alpha Subunit / genetics
-
Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
-
Hypoxia-Inducible Factor-Proline Dioxygenases / genetics
-
Hypoxia-Inducible Factor-Proline Dioxygenases / metabolism
-
Indazoles / pharmacology
-
Ligustrum / chemistry*
-
Plant Extracts / chemistry
-
Pyrans / isolation & purification
-
Pyrans / pharmacology*
-
Rats
-
Rats, Sprague-Dawley
-
Retinal Neovascularization / drug therapy*
-
Retinal Neovascularization / etiology
-
Retinal Neovascularization / genetics
-
Retinal Neovascularization / pathology
-
Retinal Pigment Epithelium / cytology
-
Retinal Pigment Epithelium / drug effects
-
Retinal Pigment Epithelium / metabolism
-
Signal Transduction
-
Vascular Endothelial Growth Factor A / antagonists & inhibitors*
-
Vascular Endothelial Growth Factor A / genetics
-
Vascular Endothelial Growth Factor A / metabolism
Substances
-
Angiogenesis Inhibitors
-
Glucosides
-
HIF1A protein, human
-
Hypoxia-Inducible Factor 1, alpha Subunit
-
Indazoles
-
Plant Extracts
-
Pyrans
-
VEGFA protein, human
-
Vascular Endothelial Growth Factor A
-
3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole
-
nuezhenide
-
Cobalt
-
EGLN1 protein, human
-
Hypoxia-Inducible Factor-Proline Dioxygenases
-
cobaltous chloride