CAJ | 학술논문

用MTT测定、琼脂糖凝胶电泳和流式细胞术等方法, 观察了一氧化氮供体硝普钠(sodium nitroprusside, SNP)对体外培养的肺成纤维细胞增殖和凋亡以及Bcl-2、 Bax和p53蛋白含量的影响.结果发现: MTT吸光度、细胞数和增殖指数(proliferation index, PI)均较对照增加; 凋亡细胞数也增加, 但尚不足以出现明显的"梯形”凋亡电泳条带; 同时, 细胞内Bcl-2蛋白下调和Bax蛋白上调; 而细胞内p53蛋白含量无明显变化.结果表明, 外源性NO有增强肺成纤维细胞增殖和凋亡的双重作用, 但以促细胞增殖为主; 此作用的分子机制与Bcl-2蛋白下调和Bax蛋白上调有关.
용MTT측정、경지당응효전영화류식세포술등방법, 관찰료일양화담공체초보납(sodium nitroprusside, SNP)대체외배양적폐성섬유세포증식화조망이급Bcl-2、 Bax화p53단백함량적영향.결과발현: MTT흡광도、세포수화증식지수(proliferation index, PI)균교대조증가; 조망세포수야증가, 단상불족이출현명현적"제형”조망전영조대; 동시, 세포내Bcl-2단백하조화Bax단백상조; 이세포내p53단백함량무명현변화.결과표명, 외원성NO유증강폐성섬유세포증식화조망적쌍중작용, 단이촉세포증식위주; 차작용적분자궤제여Bcl-2단백하조화Bax단백상조유관.
The effects of nitric oxide (NO) donor sodium nitroprusside (SNP) on the proliferation and apoptosis and on Bcl-2, Bax and p53 proteins of pulmonary fibroblasts were investigated by using MTT cleavage assay, agarose gel electrophoresis and flow cytometric analysis. The results showed increases in the optical density (550 nm) of MTT cleavage assay, the number of cells and the proliferation index (PI), in comparison with the control. The number of apoptotic cells was also increased, though the percentage of apoptotic cells was too low to reveal oligonucleosomal fragmentation of characteristic ladder pattern, which is associated with apoptosis. In the meantime, the level of Bcl-2 decreased and that of Bax increased, while the p53 remained unchanged. These results suggest that exogenous NO has a dual effect on proliferation and apoptosis; and the action of NO on pulmonary fibroblasts is mainly proliferative. Down-regulation of Bcl-2 and up-regulation of Bax are implicated in the molecular mechanisms of this action.