Regulating Mechanism Of UPA And UPAR On Proliferation Of Bovine Cumulus-granulosa Cells Cultured In Vitro

The lab preliminary study shows that uPA combined with cytomembrane exterior uPAR could promote celled proliferation.For understand the specific mechanism which is about uPA and uPAR adjust culture in vitro bovine cumulus granulosa cell proliferation,this study systematically discusses uPA,uPAR,cAMP,phosphorylation ERK and AKT probable interaction mechanism in the process of bovine cumulus granulosa cell proliferation compounded cell culture,cell proliferation,Western bot,ELISA and some other technologies.The research results show that: firstly 0.5 ng/mL uPA react in 2 min,5 min,10 min,20 min,30 min,5 min treatment group's ERK and AKT phosphorylation is the highest,which overtop control group obviously(P< 0.1).When 0.5 ng/mL uPA react with 1?g/mL Amiloride for 5 min,the united added group(uPA+Amiloride)phosphorylation ERK and AKT expression quantity reduce obviously(P<0.01)compared with uPA team.It declares that uPA combined with Amiloride can restrain uPA's auxo-action for phosphorylation ERK and AKT of bovine cumulus granulosa cell.Secondly,after add 0.5 ng/mL uPA and 10?L/mL uPAR 5 min,phosphorylation ERK and AKT of united added group(uPA+uPAR antibody)reduce obviously(P<0.01)compared with uPA group.It illustrates that uPA combined with receptor uPAR could activate ERK and AKT signal.Thirdly,after add uPA and Amiloride or uPA and uPAR 5 min,added uPA group can improve cAMP's level(P<0.01)compared with control group.However,the united added group(uPA+Amiloride or uPA+uPAR antibody)reduce cAMP's level obviously compared with uPA group(P<0.01).It shows that Amiloride can offset the impact which uPA promotes bovine cumulus granulosa cell of c AMP's level and it is related to uPAR.Fourthly,after add 0.5 ?M Rp-cAMP and uPA 5 min,untied added group(uPA+ Rp-cAMP)phosphorylation ERK and AKT's expression quantity reduce obviously(P<0.01)compared with uPA group.It explains that uPA activate backward position ERK and AKT phosphorylation through turning up cAMP level and stimulating PKA signal channel.Fifthly,after unite add Rp-cAMP and uPA 48 h,compared with uPA group,Bovine cumulus granules' cell number of the united added group(uPA+Rp-cAMP)reduce obviously(P<0.01).It shows that Rp-cAMP could offset uPA's entrainment for bovine cumulus granulosa cell proliferation.In conclusion,the study reveals that uPA and uPAR is the signaling mechanism of adjusting bovine cumulus granulosa cell proliferation.uPA combined with receptor uPAR in Bovine cumulus granules cytomembrane improve cAMP level,activate PKA passageway and phosphorylate backward position ERK and AKT.Finally it promotes Bovine cumulus granules' cell proliferation which cultivated in vitro.The results of this study are of great significance for understanding the regulation mechanism of bovine follicular development.