title: A New Recombinant PACAP-Derived Peptide Efficiently Promotes Corneal Wound Repairing and Lacrimal Secretion.
abstract: OBJECTIVE
  A new recombinant pituitary adenylate cyclase-activating polypeptide (PACAP)-derived peptide, MPAPO, which has higher stability and PAC1-specific potency, was generated. The actions of MPAPO on corneal wound repairing and lacrimal secretion were examined.
  
  
  METHODS
  MPAPO was prepared and identified by gene recombination, high-performance liquid chromatography (HPLC), and electrospray ionization mass spectrometry (ESI-MS). Stability assay was performed by HPLC-ESI-MS. PAC1-specific binding and potency assays were performed using PAC1-CHO cells. C57BL/6 mice and Japanese white rabbits were respectively used to analyze the effects of MPAPO on corneal wound repairing and lacrimal fluid secretion. Tetrazolium-based colorimetric assay (MTT), immunofluorescence, gene microarrays, and Western blot assay were performed to measure the effects of MPAPO on corneal epithelial cell proliferation, synapse growth, and gene differential expression of trigeminal ganglion cells.
  
  
  RESULTS
  As compared with the wild PACAP, the in vitro stability and PAC1-specific potency of MPAPO with four mutations (M17L, L27K and M, K, respectively, added to the N- and C-terminus) were increased approximately 31- and 2-fold, respectively. MPAPO can significantly promote the proliferation of mouse corneal epithelium cells and the synapse growth of trigeminal ganglion cells. In experimental animals, MPAPO performed a complete corneal epithelial wound closure in 30 hours and significantly inhibited corneal neovascularization, and the effects were obviously stronger than for wild PACAP and recombinant bovine (rb)-bFGF (an anti-corneal wound drug). Furthermore, MPAPO can increase the lacrimal secretion, which may efficiently improve dry eye.
  
  
  CONCLUSIONS
  MPAPO may represent a promising external therapeutic peptide for corneal wound repairing or dry eye.
authors.affiliation: Institute of Biomedicine and Department of Cellular Biology National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou, Guangdong Province, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Institute of Biomedicine and Department of Cellular Biology National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou, Guangdong Province, China.
authors.forename: Shaojun
authors.initials: S
authors.lastname: Zhao
authors.affiliation: Institute of Biomedicine and Department of Cellular Biology National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou, Guangdong Province, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Biomedicine and Department of Cellular Biology National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou, Guangdong Province, China.
authors.forename: Shutao
authors.initials: S
authors.lastname: Shen
authors.affiliation: School of Medicine of Jinan University, Guangzhou, Guangdong Province, China.
authors.forename: Min
authors.initials: M
authors.lastname: Ma
authors.affiliation: Institute of Biomedicine and Department of Cellular Biology National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou, Guangdong Province, China.
authors.forename: Wenna
authors.initials: W
authors.lastname: Xu
authors.affiliation: Institute of Biomedicine and Department of Cellular Biology National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou, Guangdong Province, China.
authors.forename: An
authors.initials: A
authors.lastname: Hong
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D065306:Corneal Injuries / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D005260:Female; D007765:Lacrimal Apparatus / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide / Q000494:pharmacology*; D011817:Rabbits; D011994:Recombinant Proteins; D013666:Tears / Q000378:metabolism*; D014945:Wound Healing / Q000187:drug effects*
chemical_list: C495402:Adcyap1 protein, mouse; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D011994:Recombinant Proteins