摘要
To explore the relationship between senescent pulmonary microvascular endothelial cells (PMVECs) in-duced by D-galactose (D-gal) and apoptosis, PMVECs were cultured with DMEM containing D-gal (10 g/L) and identi-fied by the following methods. We found that about 90% senescent cells were b-galactosidase positive cells, and 90% cells entered an irreversible G1 growth arrest. The cells in S and G2/M phases nearly disappeared. These results indi-cated that PMVECs ageing model induced by D-gal was es-tablished 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 in-duced by D-gal were sensitive to apoptosis. Apoptosis is the regular feature of senescent PMVECs.
To explore the relationship between senescent pulmonary microvascular endothelial cells (PMVECs) in-duced by D-galactose (D-gal) and apoptosis, PMVECs were cultured with DMEM containing D-gal (10 g/L) and identi-fied by the following methods. We found that about 90% senescent cells were b-galactosidase positive cells, and 90% cells entered an irreversible G1 growth arrest. The cells in S and G2/M phases nearly disappeared. These results indi-cated that PMVECs ageing model induced by D-gal was es-tablished 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 in-duced by D-gal were sensitive to apoptosis. Apoptosis is the regular feature of senescent PMVECs.
基金
supported by the National Major Basic Research Program in China(Grant No.G2000057004)
the National Natural Science Foundation of China(Grant No.30170400).
