The Function Change Of Human Umbilical Vein Endothelial Cells Affected By Recombinant Adeno-associated Virus-mediated Kallikrein Gene

Objective To evaluate the variance of quantity of human tissue kallikrein (HTK) synthesized by HUVEC cells which are infected with constructed rAAV-HTK and the functional changes of HUVEC cells.To discuss the feasibility of treating hypertension and ischemic heart disease with rAAV-HTK.Methods 1.HUVEC cells were infected by constructed rAAV-HTK. 2. Measure the quantity of HTK in the HUVEC and the medium by ELISA. 3. Measure the variance of mRNA quantity of eNOS, caspase-3, ET-1, VEGF , ETR-B,Bradykinin B₁ receptor and Bradykinin B₂ receptor in the HUVEC cells before and after infection by semiquantitative RT-PCR.Results 1.The quantity of HTK in the HUVEC cells infected with rAAV-HTK is three times than the control. 2. HTK can not be detected in the medium of the HUVEC cells. 3. The mRNA quantity of eNOS in the infected HUVEC cells is higher than the control,but the mRNA quantity of caspase-3 is lower, and there is no obvious difference with VEGF,ET-1,ETR-B,Bradykinin B1 receptor and Bradykinin B2 receptor.Conclusions More HTK are synthesized by the HUVEC cells infected with the rAAV-HTK. The rAAV-HTK infected HUVEC cells synthesize more HTK, increase the the quantity of eNOS mRNA and NO, decrease the quantity of caspase-3 mRNA, which implies the HTK can enhance endothelial cells dysfunction. These beneficial effects show thepotential implication for the gene therapy in treatingcardiovascular deseases with HTK gene caused by blood vesselendothelium dysfunctuon,such as hypertesion et al.