title: PAC1R agonist maxadilan enhances hADSC viability and neural differentiation potential.
abstract: Pituitary adenylate cyclase-activating polypeptide (PACAP) is a structurally endogenous peptide with many biological roles. However, little is known about its presence or effects in human adipose-derived stem cells (hADSCs). In this study, the expression of PACAP type I receptor (PAC1R) was first confirmed in hADSCs. Maxadilan, a specific agonist of PAC1R, could increase hADSC proliferation as determined by Cell Counting Kit-8 and cell cycle analysis and promote migration as shown in wound-healing assays. Maxadilan also showed anti-apoptotic activity in hADSCs against serum withdrawal-induced apoptosis based on Annexin V/propidium iodide analysis and mitochondrial membrane potential assays. The anti-apoptotic effects of maxadilan correlated with the down-regulation of Cleaved Caspase 3 and Caspase 9 as well as up-regulation of Bcl-2. The chemical neural differentiation potential could be enhanced by maxadilan as indicated through quantitative PCR, Western blot and cell morphology analysis. Moreover, cytokine neural redifferentiation of hADSCs treated with maxadilan acquired stronger neuron-like functions with higher voltage-dependent tetrodotoxin-sensitive sodium currents, higher outward potassium currents and partial electrical impulses as determined using whole-cell patch clamp recordings. Maxadilan up-regulated the Wnt/β-catenin signalling pathway associated with dimer-dependent activity of PAC1R, promoting cell viability that was inhibited by XAV939, and it also activated the protein kinase A (PKA) signalling pathway associated with ligand-dependent activity of PAC1R, enhancing cell viability and neural differentiation potential that was inhibited by H-89. In summary, these results demonstrated that PAC1R is present in hADSCs, and maxadilan could enhance hADSC viability and neural differentiation potential in neural differentiation medium.
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Guo
authors.affiliation: Department of Cell Biology, Jinan University, Guangzhou, China.
authors.forename: Rongjie
authors.initials: R
authors.lastname: Yu
authors.affiliation: GHM Institute of CNS Regeneration, Jinan University, Guangzhou, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Ophthalmology, The First Clinical Medical College of Jinan University, Guangzhou, China.
authors.forename: Ruiling
authors.initials: R
authors.lastname: Lian
authors.affiliation: GHM Institute of CNS Regeneration, Jinan University, Guangzhou, China.
authors.forename: Yankun
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Cell Biology, Jinan University, Guangzhou, China.
authors.forename: Zekai
authors.initials: Z
authors.lastname: Cui
authors.affiliation: Department of Ophthalmology, Affiliated Anhui Provincial Hospital of Anhui Medical University, Hefei, China.
authors.forename: Qingshan
authors.initials: Q
authors.lastname: Ji
authors.affiliation: Department of Mathematics, South China University of Technology, Guangzhou, China.
authors.forename: Junhe
authors.initials: J
authors.lastname: Chen
authors.affiliation: Eye Institute, Medical College of Jinan University, Guangzhou, China.
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Li
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Liu
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China.|Department of Ophthalmology, The First Clinical Medical College of Jinan University, Guangzhou, China.|Eye Institute, Medical College of Jinan University, Guangzhou, China.
authors.forename: Jiansu
authors.initials: J
authors.lastname: Chen
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D017304:Annexin A5 / Q000378:metabolism; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053453:Caspase 9 / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology; D006801:Humans; D019476:Insect Proteins / Q000494:pharmacology*; D007546:Isoquinolines / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018408:Patch-Clamp Techniques; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D051237:Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology; D013779:Tetrodotoxin / Q000494:pharmacology; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism
chemical_list: D017304:Annexin A5; C000595707:BCL2 protein, human; C495270:CTNNB1 protein, human; D006575:Heterocyclic Compounds, 3-Ring; D019476:Insect Proteins; D007546:Isoquinolines; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D019253:Proto-Oncogene Proteins c-bcl-2; D051237:Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I; D013449:Sulfonamides; D051153:Wnt Proteins; C544261:XAV939; D051176:beta Catenin; C069147:maxadilan protein, insect; D013779:Tetrodotoxin; D017868:Cyclic AMP-Dependent Protein Kinases; C505407:CASP3 protein, human; C506433:CASP9 protein, human; D053148:Caspase 3; D053453:Caspase 9; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide
keywords: apoptotic; hADSCs; maxadilan; neural differentiation; proliferation