Lentiviral-AQP1 vector successfully transfect the myocytes and aggravate the myocardiac edema after cardiopulmonary bypass / 中华胸心血管外科杂志

ABSTRACT

Objective Construct the lentiviral AQP1 vector and explore whether it can transfect the myocyte or not,then test the law of the AQP1 expression and the edema in the successfully transducted myocytes after cardiopulmonary bypass in sheeps.Methods Design cleavage primer according to ovine AQP1mRNA,clone it into expressing vector and transducated into the 293T cells with other packing vectors to produce the lentiviral AQP1 vector,then test the virus titer.36 adult healthy sheeps are randomly divided into blank or AQP1-lentiviral transfected group,blank or AQP1-lentiviral vector suspension was injected in the ventricle tissue of healthy adult sheeps during cardiopulmonary bypass and take specimen in different time points (2,6,12,24,48,72 h)after extracorporeal circulation,3 in each group.Realtime-PCR WesternBlot ELISA FACS immumofluorescent and Dry/Wet methods are emploied to detect the expression of AQP1 and the according degree of edema.Results lentiviral AQP1 vector was successfully construced and transducated into the myocytes.The tranducated groups have the same trend of AQP1 of expression and cardiac edema after cardiopulmonary bypass compared to the blank vector group,but the degree is heavier(P ＜ 0.05).Conclusion Lentivral AQP1 vector can successfully transfect the myoctyes,and the overexpressed myocardial tissue have the same trend of AQP1 expression and edema after cardiopulmonary bypass,but with a heaver degree.The expression of aquaporins was positively relevant to the edema.