Activated protein C (APC) is serine protease hemostasis, independent of its anticoagulant activity, exhibits anti-inflammatory and anti-apoptotic properties that determine the possibility of the protective effects of APC in different diseases, including sepsis and chronic wound healing. APC, binding of endothelial protein C receptor (EPCR) and specifically cleaving PAR1 receptor and releasing peptide agonist PAR1 stabilizes not only endothelial cells, but also many others, including epidermal keratinocytes of the skin. We develop the hypothesis that the cytoprotective effect of APC on the cells, involved in wound healing, seem to imitate peptide - analogous of PAR1 "tethered ligand" that activate PAR1. In our work, we synthesized a peptide (AP9) - analogue of PAR1 tethered ligand, released by APC, and firstly showed that peptide AP9 (0.1-10 мM), like to APC (0.01-100 nM), stimulates the proliferative activity of human primary keratinocytes. Using a model of the formation of epithelial wounds in vitro we found that peptide AP9, as well as protease APC, accelerates wound healing. Using specific antibodies to the receptor PAR1 and EPCR was studied the receptor mechanism of AP9 action in wound healing compared with the action of APС. The necessity of both receptors - PAR1 and EPСR, for proliferative activity of agonists was revealed. Identified in our work imitation by peptide AP9 - PAR1 ligand, APC acts on keratinocytes suggests the possibility of using a peptide AP9 to stimulate tissue repair.
Aktivirovannyĭ protein S (APS) – serinovaia proteaza gemostaza, nezavisimo ot antikoaguliantnoĭ aktivnosti proiavliaet protivovospalitel'nye i antiapoptoticheskie svoĭstva, kotorye obuslavlivaiut vozmozhnost' protektornogo deĭstviia APS pri raznykh zabolevaniiakh, vkliuchaia sepsis i zazhivlenie khronicheskikh ran. Predpolozhiv, chto tsitoprotektornoe deĭstvie APS na kletki, uchastvuiushchie v zazhivlenii ran, mozhet byt', po-vidimomu, imitirovano peptidami – analogami “priviazannykh ligandov”, kotorye aktiviruiut proteazami aktiviruemyĭ retseptor 1 tipa (PAR1), my sintezirovali peptid (AP9) – analog priviazannogo liganda peptida agonista PAR1, osvobozhdaemogo APS. V rabote vpervye pokazano, chto peptidAP9 (0,1- 10 mkM), podobno proteaze APS (0,01-100 nM), stimuliruet proliferativnuiu aktivnost' pervichnykh keratinotsitov cheloveka. V modeli rany épitelial'nogo plasta peptid AP9, takzhe kak proteaza APS, uskoriaet zakrytie rany. S pomoshch'iu spetsificheskikh antitel byl izuchen retseptornyĭ mekhanizm deĭstviia AP9 pri zazhivlenii rany v sravnenii s deĭstviem APS. Pokazana neobkhodimost' retseptora PAR1 i éndotelial'nogo retseptora proteina S (ÉPSR) dlia proliferativnoĭ aktivnosti agonistov. Vyiavlennaia v nasheĭ rabote imitatsiia peptidom 9 - ligandom PAR1, deĭstviia APS na keratinotsity ukazyvaet na vozmozhnost' ispol'zovaniia peptida AP9 dlia stimuliatsii reparatsii tkaneĭ.
Keywords: keratinocytes; peptide agonist; protease-activated receptor; protein C; wound healing.