[A synthetic peptide selected by bioinformatics inhibits mouse corneal neovascularization].
OBJECTIVE
To identify the active anti-angiogenic region in the amino acid sequence of human apolipoprotein (a) [apo (a)] kringle V (KV), and to evaluate the role of this synthetic peptide on VEGF-induced angiogenesis of mouse cornea in vivo.


METHODS
The characterization of the structure and biological activity of the amino acid sequence of apo (a) KV was analyzed using the bioinformatic methods which included sequence alignment, analysis of antigenicity, surface accessibility and hydrophilicity, and then a peptides was selected. The peptide was synthesized with a high efficiency solid-phase method. Corneal neovascularization was induced with a pellet containing 160 ng vascular endothelial growth factor (VEGF) in a mouse corneal micropocket model. 40 C57BL/6 mice (40 eyes) were divided randomly into 4 groups (10 eyes per group). Four kinds of pellets were made containing 160 ng VEGF plus the dose range of 0.0, 0.5, 1.0 and 1.5 microg synthetic peptide for control group, group A, group B and group C, respectively. Neovascularization was observed biomicroscopically on day 7 after the operation, and the corneas were then examined histologically.


RESULTS
The result of bioinformatic analysis showed that the peptide contained a majority of conservative residues and possessed fine properties of antigenicity, surface accessibility and hydrophilicity. The synthetic peptide at the doses of 1.0 microg and 1.5 microg showed significant inhibition of mouse corneal neovascularization induced by VEGF in the parameters of vessel length, clock hours and area compared with the control group on day 7 after the operation (P < 0.01). There was no difference in the two doses (1.0 microg and 1.5 microg peptide) in the inhibition of the neovascularization. The dose of 0.5 microg peptide did not show any significant inhibition of the neovascularization compared with the control group (P > 0.05).


CONCLUSIONS
The peptide, selected from the amino acid sequence of apo (a) KV by bioinformatics, appears to inhibit VEGF-induced angiogenesis in a mouse corneal micropocket assay in vivo, therefore, the study suggest that this amino acid sequence may locate at the active anti-angiogenic region of apo (a) KV.
Zhao
Hui
H
Department of Ophthalmology, the First People's Hospital, Jiaotong University, Shanghai 200080, China.
Yi
Zheng-fang
ZF
Hu
Hong-hui
HH
Xu
Xun
X
D000595:Amino Acid Sequence; D000818:Animals; D001054:Apolipoproteins A / Q000235:genetics* / Q000302:isolation & purification / Q000627:therapeutic use; D019295:Computational Biology; D016510:Corneal Neovascularization / Q000188:drug therapy; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010455:Peptides / Q000235:genetics* / Q000302:isolation & purification / Q000627:therapeutic use; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D001054:Apolipoproteins A; D010455:Peptides; D042461:Vascular Endothelial Growth Factor A