Effects Of CyclinD、CDK4 And PRb Phosphorylation Status By Method Of Bu Shen Sheng Sui Combined With BMSCs Transpl Antion On D-Galactose Induced Aging Rats

Objective: This study focuses on the effect of BuShenShengSuiFang combined with Bone marrow mesenchymal stem cells(BMSCs) transplantion on the D-galactose induced aging rat on cyclinD 、 cyclin-dependent protein kinases4( CDK4)、 state of phosphorylation of the gene expression product of Retinoblastoma-pRb,to explore the role and mechanisms of Method of BuShenShengSui combined with Bone marrow mesenchymal stem cells transplantion for anti aging.Methods:(1)Preparing the subacute aging rat model by subcutaneous injecting the 10%D–galactose on nape,by contrasting the aging degree scores and determining the Superoxide Dismutase(SOD)activity and Malondialdehyde malonic dialdehyde Propanedial(MDA)content of rat serum to judge preparation success in aging rat model between with natural aging rats group, youth group of rats.(2)By density gradient centrifugation combined with adherence separation screening method in vitro to cultivate BMSCs.(3)Dividing the subacute aging rat model into four parts:Group Ⅰ( Model group)、 Group Ⅱ( BuShenShengSuiFang group)、 Group Ⅲ( BuShenShengSuiFang+BMSCs transplantion group)、 Group Ⅳ( BMSCs transplantion group).After made the appropriate intervention,to contrast the aging degree scores of each group,and determine the SOD activity and MDA content of rat serum,and detect the cyclinD、CDK4 and state of phosphorylation of pRb of rat hippocampal tissue by immunohistochemical method.Results:(1)In subacute aging rat model group,each aging degree score rose,the SOD activity decreased significantly,and the level of MDA increased also,there was no statistically significant difference compared with natural aging rats group(P>0.05), and there was statistically significant difference compared with youth group of rats(P<0.05),and the subacute aging model was successful prepared.(2)Through the morphological observation, primary cultured BMSCs is adherent growth with long fusiform shape or polygon in a single,and colony distribution.P3,P4,P5 generation of BMSCs growth curve approximate "S" shape.The positive rate of P3 BMSCs Surface markers CD71、CD44 was above 95%, Subculture cell conform to the requirements of the experiments.(3)After different intervention, there was statistically difference between the groups in the aging degree scores, the SOD activity, the MDA content and the positive rate of cyclinD、CDK4 and state of phosphorylation of pRb(P<0.05). Two comparison in the three groups,compared with the Group Ⅰ,the aging degree scores of Group Ⅱ、Group Ⅲ、Group were falling,the SOD activity Ⅳwere ascent,and the MDA content were descend,and there was statistically difference between the groups(P<0.05). Comparison between the Group Ⅱ、Group Ⅲand Group Ⅳ, there was statistically difference between the groups(P<0.05),and among them,the rise of SOD activity and decline of MDA content in Group were the most Ⅲobvious, the positive rate of cyclinD、CDK4 and state of phosphorylation of pRb were respective 90%、90%、80%,there was statistically significant difference between the groups(P<0.05).Conclusion:Method of BuShenShengSui combined with Bone marrow mesenchymal stem cells transplantion could effectively anti aging,which may play a role by conduction and control on cyclinD-CDK4-pRb pathway in cells.