The Study Of Exogenous Retinoic Acid On Myocardial Cell Apoptosis

Objective:To explore the influence of exogenous retinoic acid (RA) on apoptosis in lactation rats myocardial cell and cellular control factor paired box gene8(Pax-8)、Fas、Fas1、Casepase3expression level. Methods:1. Cultivate myocardial cells and defining the best time of intervening in rat cardiomyocytes:Under the aseptic conditions, Take out the heart from Balb/c milk rat, Using trysin digestion-differential and purification for myocardial cells and to cultivate myocardial cells.2.The determination of exogenous retinoic acid on myocardial cell apoptosis:Set blank control group,Myocardial cells were cultured in20%DMEM. In different concentrations depending on the retinoic acid (0.5,1.0,2,4umol/L) to intervene. After0、12、24、48hour, we measured the inhibition rate of the myocardial cells with MTT technique; cell apoptosis with flow cytometry.3.The determination of Cellular control factor:We measured the expression of Fas,Fas,Caspase-3, Pax-8protein by using Western blot technique; We measured the expression of Fas,Fas mRNA by using RT-PCR technique. Results:1. In72hour, it is in the middle of the exponential phase,in which cells’ growth was the most vigorously and is the optimal time interfering and observation of the effect of which.2. At higher concentration (2.0umol/L)RA could damage celular morphology and activities,increase apoptosis. At the same time, the higher the concentration, the more time, the inhibition efficiency is also more obvious.3. Compared with the control,2umol/L depending on the yellow acid after intervention with the role the extension of time,The expression of Fas、Fas1、Caspase-3protein are higher gradually (P<0.05); Instead of Pax-8expression then gradually reduce (P<0.05). The expression of Fas、Fas1mRNA are higher significantly (P<0.05).Conclusion:1.Using trypsin digestion-differential adhesion and5-bromo deoxyuridine inhibiting methods can successfully cultured neonatal rat cardiomyocyte; In72hour, It is the optimal time interfering and observation of the effect of which.2.At higher concentration RA could damage celular morphology and activities,increase apoptosis.3. Exogenous retinoic acid could inhibit proliferation and promote the apoptosis of myocardial cells by adjusting the transcription factors Fas、Fas1、 Caspase3、Pax-8expression.