The Study On The Induction Of Apoptosis By Egb In Cultured Fetuses Vascular Smooth Muscle Cell

Objective Investigate the induction effect of Egb on apoptosis of VSMCs cultured from the thoracic aortas of the aborted fetus in vitro.Methods Culture VSMCs from the thoracic aortas of the aborted fetus by Collagenase IV digested combined with tissue anchored. Add uptitratal concentrations of Egb(Oμg/ml, 200 μg/ml, 400 μg/ml, 800 μg/ml) to the culture medium of cells which grew well at 5-8 passages. At the different end time point (6h, 12h, 24h) , We observed apoptosis phenomena and detected the ratio of early phase apoptosis cells labled with Annexin- V combined PI by flow cytometry. We also detected Fas and Bcl-2 which were correlated protein with apotosis by flow cytometry.Results Observed under inverted phase contrast microscope, compared with the control group,cells in Egb group shrinked, turned smaller round and budded around the cell member. Stain with Gimsa and observed with electron microscope may found the apoptosis phenomenon such as apoptotic body and so on.Detected with FCM: We found along with the increase of concentration and exposed time,the apoptosis rate was increased. At 800μg/ml Egb concentration and at 24h end point,the apoptosis rate rised to 34.4%.Statistical analysis showed along with the increased of Egb concentration and exposed time, the apoptosis rate was increased. And it showed obviously statistical significance. We also found that along with the increase of Egb concentration and the prolong of exposed time, the expression of Fas was increased and the expression of Bcl-2 was descended.(p=0.00)Conclusions Egb may induce apoptosis of VSMC . Along with the increased of Egb concentration and exposed time, the apoptosis rate was increased. With the gain of apoptosis rate, the expression of Fas increased and expression of Bcl-2 descended.