Objective:To construct PI3-K/Akt shRNA expression vectors. Methods:Eight 19~21 bp reverse repeated motifs targeting of PI3-K/Akt gene were synthesized and cloned into eukaryotic expression plasmid pGenesil-1. After being screened and sequencing confirmed,the recombinant plasmids were transfected into rat glomerular mesangial cells(GMCs),then the levels of P-Akt and T-Akt protein in rat GMCs were measured using Western blot to select the optimal shRNA. Results:It was verified by partial nucleotide sequencing and restriction endonuclease digestion that the constructed eukaryotic vector expressing shRNA of PI3-K/Akt were correct. Western blotting results showed that the optimal shRNA which could effectively silence the target genes were shPik3c3-2 and shAkt1-4 respectively. Conclusion:The eukaryotic vectors expressing shRNA of PI3-K/Akt were constructed successfully.