The Effect Of Pseudolaric Acid B On Proliferation Of Human Keloids Fibroblasts And The Expression Of PI3K/AKT?COX-2?caspase-3 In Vitro

Objective To investigate the effect of pseudolaric acid B on proliferation of Human keloid fibroblasts and the expression of PI3K/AKT?COX-2?caspase-3 in vitro before and after intervention of the keloid fibroblasts and explore its m echanism.Methods(1)Fibroblasts identification: Preparation the climbing cells film, giving the HE staining and using anti-vimentin antibodies for cell identification by t he streptavidin-perosidase(SP) immunohistochemical staining techniques.(2)With different concentrations of pseudolaric acid B effect on proliferation of Human keloid fibroblasts cultured in vitro, and adopted to evaluate cell survival by C CK-8 assay.(3)The expression of PI3K/AKT?COX-2?caspase-3 before and aft er intervention were detected by RT-PCR and Western blot.(4)Then use statist ical software for data processing and statistical analysis.Results(1)It shows the positive expression of vimentin,and cytokeratin staining was negative by the SP immunohistochemical staining techniques. The positive expression of vimentin in the cell cytoplasm, its colour is brown, showing the long axis of the same cell bundle or mesh node.(2)Pseudolaric acid B affect thekeloid fibroblasts in different concentrations(0.5 umol/L, 1 umol/L, 2 umol/L,4umol/L) and different time(24h,48 h,72h), we detected the absorbance decrea sed with the concerntrations and time increased.(3)RT-PCR and western blot re sults showed that gene expression and protein expression of PI3K?AKT of th e keloid fibroblasts and fibroblasts from normal skin were all decreased,the CO X-2 of keloid fibroblasts were decreased,but gene expression and protein expr ession of caspase-3 was increased in 2 umol/L Pseudolaric acid B of Human Keloids Fibroblasts and fibroblasts from normal skin. There were significant diffe rences between these groups(P<0.05).Conclusions(1)Pseudolaric acid B significantly inhibited the proliferation o f Human Keloids Fibroblasts cell in vitro.(2)Pseudolaric acid B can inhibite the expression of PI3K?AKT and COX-2 and increase the expression of ca spase-3 in Human Keloids Fibroblasts.(3)The COX-2?PI3K/AKT?Caspase-3 si gnaling pathways may be the mechanism of Pseudolaric acid B.