Effects of cylin-dependent kinase 5 expression inhibition on apoptosis of mouse podocyte / 重庆医学

ABSTRACT

Objective To investigate the expression of Cdk5 in high glucose-induced mouse podocytes and the effects of inhibi-ting Cdk5 expression on podocyte apoptosis .Methods (1)Mouse podocytes cultured in vitro were divided into normal glucose group ,mannitol group ,high glucose group and the high glucose group was further divided in to 0 ,6 ,12 ,24 ,48 h gorup depend on the induce time .Changes of Cdk5 expression in each group were detected by Western blotting .(2)The expression of Cdk5 in podo-cyte was inhibited by Cdk5 miRNA plasmid .The podocytes were divided into 4 groupsNG group(normal glucose) ,HG group(high glucose) ,HG + S group(high glucose + scrambled plasmid) and HG + C group(high glucose + Cdk5 miRNA plasmid) .The podo-cyte apoptosis rates were detected by flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) , and the expressions of Bcl-2 ,Bax and cleaved caspased-3 were detected by Western blotting respectively after 48 h .Results Com-pared with normal glucose group ,the expression of Cdk5 was significantly increased in podocytes of high glucose group (P< 0 .05) . The expression of Cdk5 in podocytes was inhibited by Cdk5 miRNA plasmid .Compared with HG and HG + S group ,the podocyte apoptosis rate of HG + C group was significantly decreased(P< 0 .05) .The expression of cleaved caspase-3 protein and Bax /Bcl-2 ratio were also decreased (P< 0 .05) .Conclusion High glucose could increased the expression of Cdk5 in podocytes .The downregu-lation of Cdk5 expression could decrease the podocyte apoptosis induced by high glucose .