Ki67和p53蛋白表达在外源性硫化氢抗内皮细胞老化过程中的作用

Role of Ki67 and p53 in the exogenous Hydrogen sulfide delaying the rat aortic endothelial cells aging

目的探索外源性硫化氢（H2S）抗内皮细胞老化过程中对Ki67和p53表达量的影响。方法将大鼠主动脉内皮细胞（RAECs）随机分为3组，即年轻组（原代RAECs传至第4代）、老年组（原代RAECs传至第12代）、硫氢化钠（NaHS，细胞内H2S的供体，即外源性H2S）组（RAECs从第4代开始给予NaHS10μmol／L处理至第12代）。用β-半乳糖苷酶（SAβ-gal）染色法鉴定细胞老化程度，DAPI荧光染色检测老化相关的异染色质位点（SAHF）的表达变化，Western blot方法检测Ki67及p53蛋白的表达量。结果给予NaHS处理后，细胞老化程度及SAHF的表达明显低于老年组（P〈0．01）；Ki67及p53蛋白在老年组RAECs中表达量明显低于年轻组（P〈0．01），而NariS处理后Ki67及p53表达量较老年组明显增加（P〈0．05）。结论外源性H2S（即NaHS）延缓内皮细胞老化过程中Ki67及p53基因的表达显著增加，提示外源性H2S延缓血管内皮细胞老化可能与提高Ki67、p53表达有关。

Objective To investigate the expression of Ki67 and p53 in the exogenous administration of hydrogen sulfide （H2S）delaying the rat aortic endothelial cells （RAECs） senescence. Methods Cultured RAECs at the fourth passage was taken as young group, and the twelfth passage was taken as old group. NariS group was incubated with sodium hydrosulphide （NariS, exogenous hydrogen sulfide, 10 μmol/L） from the fourth to twelfth passage. The cell senes- cence status was detected by the β -galactosidase （SAβ -gal） staining, DAPI fluorescent dye was used to determine senescence associated heterochromatin foci （SAHF） formation; Western blot technique was used to analysis the expression of Ki67 and p53 at the level of protein, respectively. Results In comparison with young group, the old group exhibited higher SA[3 - gal positive rate and SAHF formation rate （P 〈 0.01 ）. Meanwhile, NariS group decreased the levels （P 〈 0.05 ）. Compared with the old group, the NariS group increased the expression of Ki67 and p53 （ P 〈 0.05 ）. Conclu- sion The expression of Ki67 and p53 is increased in the process of exogenous hydrogen sulfide delaying endothelial cell senescence. These findings suggest that the effect related to the expression of Ki67 and p53.