CD73对体外心成纤维细胞和心肌细胞增殖与衰老的影响OACSTPCD

Objective:To investigate the effects of CD73 on the proliferation and senescence of cardiac fibroblasts(CFs)and cardiomyocytes(CMs).Methods:Rats CFs and CMs were cultured in vivo.The lentiviral vector with CD73 overexpression was constructed and transfected into CFs and CMs.Experimental groups were the CD73 overexpression group(OE group)and negative control group(NC group).The gene expression of CD73 in both groups was detected by RT-PCR assay.The cell senescence was detected by senescence-associated β-galactosidase staining.The cell proliferation was detected by MTT assay.The cell growth curve and doubling time were measured by a non-invasive cell function analyzer.The cell cycle was detected by flow cytometry.The RT-PCR method was used to detect the changes in the mRNA expression of proliferation-related signal molecules.Results:Seventy-two hours after the lentiviral transfection of CFs and CMs,the levels of CD73 mRNA in the OE group were significantly higher than those in the NC group.The number of senescent CFs and CMs cells in the NC group was higher than that in the OE group.The proliferative ability of CMs after transfection for 3-4 d was higher in the OE group than that in the NC group,while there was no significant difference between the OE group and the NC group in the proliferative ability of CFs.On the 3rd day after transfection,the cell cycle changes were as follows:There was no significant difference in the proportion of CFs cells in the G1 and S phases of both groups,while the proportion of CFs cells in the G2 phase was slightly higher in the OE group than that in the NC group.The growth curves of CFs and CMs both showed that the OE group had higher growth rates than those in the NC group.The doubling time of CMs in the OE group was shorter than that of the NC group,but the difference in CFs was not significant.On the 3rd day after transfection,the expression of PI3K and Akt mRNA were increased in CMs,while the expression of vascular endothelial growth factor(VEGF)mRNA was unaffected.Conclusion:CD73 has no significant effect on the proliferation of CFs but inhibits their senescence.CD73 promotes CMs proliferation and inhibits their senescence.CD73 can promote the high expression of PI3K and Akt in CMs,but it has no effect on VEGF expression.