Abstract
Purpose: To investigate the effect of bone morphogenetic protein-2 (BMP-2) on human retinal vascular endothelial cells (RECs) and human retinal pigment epithelial cells (RPE) cultured in high glucose (HG) in vitro, and the underlying mechanism.
 Methods: Cell counting kit-8 (CCK-8) was used to determine cell proliferation while Western blot was used to assay the expressions of extracellular matrix and angiogenesis-related factors, Expressions of cytokines and chemokines were assessed by quantitative real time polymerase chain reaction (qRTPCR) and enzyme-linked immunosorbent assay (ELISA). Changes in Smad, ERK, JNK and p38MAPK signal pathway were measured by transfection and interference.
 Results: The level of expression of BMP-2 in HG group was higher than that in normal glucose (NG) culture group. The expressions of angiogenesis-related factors i.e. vascular endothelial growth factor (VEGF) and intercellular cell adhesion molecule-1 (ICAM1), pro-inflammatory factors i.e. IL-6 and chemokine monocyte chemokine protein-1 (MCP1), increased significantly in HG group compared to NG and HG + BMP-2 groups. Phosphorylation of Smad1/5/8 and activation of ERK, JNK and p38MAPK signaling pathways were enhanced by BMP-2.
 Conclusion: These results suggest that BMP-2 promotes angiogenesis and enhances the expressions of inflammatory cytokines via Smad signaling pathway.
Highlights
Diabetic retinopathy (DR) is one of the predisposing factors to blindness among people in the age range of 26 - 75 years
The expression of ZO-1 in high glucose (HG) + bone morphogenetic protein-2 (BMP-2) group was lower than that in HG group. These results demonstrated that the migration ability in the HG + Bone morphogenetic protein (BMP)-2 group was lower than that in the pure HG group (p < 0.05)
The expressions of SMA α and MMP2 in HG group was markedly higher than that in normal glucose (NG) group, and the expression of HG + BMP-2 group was significantly higher than that in HG group (p < 0.05). These results indicate that the expression of ZO-1 was inhibited by BMP-2, while the expressions of SMA α and MMP2 were enhanced by BMP-2
Summary
Diabetic retinopathy (DR) is one of the predisposing factors to blindness among people in the age range of 26 - 75 years. Bone morphogenetic protein (BMP) contains a wide range of conserved growth factor groups, more than 30 members of which have been identified to date, and it is the largest member of the transforming growth factor beta (TGFβ) superfamily [4] Three members of this family i.e. BMP-2, BMP-4 and BMP-7 and their receptors (BMPR) play important roles in ocular biochemistry. In the present study, the effect BMP-2 on retinal pigment epithelium (RPE) and retinal vascular endothelial cells (RECs) cultured in HG, and the mechanisms involved, were investigated in vitro. This was with a view to providing new scientific information that might enhance the prevention and treatment of DR. Values of p < 0.05 were considered statistically significant
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