Abstract
To explore the relationship between senescent pulmonary microvascular endothelial cells (PMVECs) induced by D-galactose (D-gal) and apoptosis, PMVECs were cultured with DMEM containing D-gal (10 g/L) and identified by the following methods. We found that about 90% senescent cells were β-galactosidase positive cells, and 90% cells entered an irreversible G1 growth arrest. The cells in S and G2/M phases nearly disappeared. These results indicated that PMVECs ageing model induced by D-gal was established successfully. Compared with the control, PMVECs induced by D-gal showed that chromatin condensation and apoptotic bodies in nucleus were detected by fluorescence microscopy and transmission electron microscopy. Apoptotic cells in the early and middle phases increased significantly (39.8 ±2.8)% vs. (14.0 ±3.7)%. Typical Sub-G1 peak in the late apoptosis phase appeared in the terminal stage of PMVECs ageing. It is concluded that D-gal induces cultured PMVECs replicative senescence. Senescent PMVECs induced by D-gal were sensitive to apoptosis. Apoptosis is the regular feature of senescent PMVECs.
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