Puerarin Through The Regulation Of Reactive Oxygen Species On Hypoxia Induced Proliferation Of Pasmcs

Objective:The paper to discuss the effect of Pue on the proliferation of hypoxia-induced pulmonary artery smooth muscle cells (PASMCs),and discuss the mechanism whether through the regulation of reactive oxygen to implementation.Methods:The original generation develop PASMCs in rats,uesd2-5generations in experiments.Using MTT to determate under different concentration gradient of Pue affect PASMCs proliferation activity.Western blot detection Pue PASMCs of rat in the tumor necrosis factor(TNF alpha)expression.Using flow cytometry instrument determination of Pue PASMCs proliferation cycle impact.Western blot detection Pue PASMCs of rat in the cell cycle protein A(CyclinA),the influence of the cell proliferating nuclear antigen(PCNA)expression.Use Pue DCFH-DA probe observation on oxygen produced by reactive oxygen species(ROS)content and the influence of Western blot detection hypoxia-inducible factor (H1F-1alpha) expression.Study the molecular mechanism of Pue influence PASMCs proliferation.According to the determined by MTT results statistical Pue drug concentrations of250mumol·L-1.ROS blocker of Rotenone(Rotenone)concentration of2.5mumol L-1,drug effect time of24h.Westernblot method is used to detect whether the ROS to participate in the Pue inhibit hypoxia induced PASMCs proliferation in the process,were determined the expression of PCNA.CyclinA and alpha HIF-1.Experiments are grouped into:oxygen.oxygen.oxygen+often Pue,anoxic+Pue+Rotenone, anoxic+Rotenone.Results:the determined by MTT results showed that hypoxia can increase the cell p roliferation activity,induction of PASMCs proliferation, and from250mumol·L-1when c oncentration of Pue have significant differences,often Pue oxygen condition of PASMCs had no effect on cell proliferation.Often oxygen and hypoxia on the expression of TN F alpha of Pue is no effect.Flow cytometry instrument test results confirmed that the P ue cells of oxygen condition often influence,and to hypoxia induced proliferation of cell s have the effect of the inhibition of the proliferation.Pue can reverse the CyclinA indu ced by hypoxia,PCNA and HIF-1alpha expression.Pue can make a significant reduction in the anoxic conditions of ROS.Rotenone and Pue have similar blocking effect on R OS.And all can suppress the PCNA,CyclinA induced by hypoxia.HIF-1alpha expression, with separate dosing Pue hypoxia or anoxia alone to join Rotenone contrast.anoxic+Pue+Rotenone group of PCNA,CyclinA,HIF-l alpha express cut more prominent.Conclusion:Pue can suppress the PASMCs proliferation induced by hypoxia.this effe ct may inhibit hypoxia under the condition of ROS generation to suppress PASMCs pr oliferation.