摘要
目的 观察FTY720对低氧性大鼠肺动脉平滑肌细胞（PASMCs）炎症通路和细胞凋亡的影响.方法 取成年雄性SD大鼠15只,用Ⅰ型胶原酶消化法进行PASMCs原代培养,将第4～6代生长良好的细胞随机分组：①常氧组（N组）;②低氧组（H组）;③P38MAPK抑制剂（SB203580）组（H＋SB组）;④～⑤：FTY720干预组（2500ng/ml、5000ng/ml）组（H＋FTY720组）,孵育24h.采用Western blot法检测总P38MAPK和总NF-κB P65以及p-P38MAPK和p-NF-κBP65的相对表达量.TUNEL法观察PASMCs的细胞凋亡情况.结果 与N组比较,H组细胞凋亡率明显降低（P＜0.05）,总P38MAPK和总NF-κB P65无明显改变（P＞0.05）,p-P38MAPK和p-NF-κB P65的表达显著升高（P＜0.01）.与H组比较,H＋FTY720组与H＋SB组的细胞凋亡率显著升高（P ＜0.05,P＜0.01）,总P38MAPK和总NF-κB P65无明显改变（P＞0.05）,p-P38MAPK和p-NF-κBP65的表达显著降低（P ＜0.05,P＜0.01）,且细胞凋亡率与FTY720浓度呈正相关（P＜0.01）.结论 在低氧条件下,PASMCs可能通过P38MAPK/NF-κB P65通路促进细胞增殖.FTY720可降低p-P38MAPK和p-NF-κBP65的表达抑制低氧性PASMCs炎症反应,抑制细胞增殖并促进细胞凋亡,缓解肺动脉高压进展.
Objective To investigate the impact of FTY720 on the apoptosis and the expression of P38MAPK and NF-κB P65 of rat hypoxie pulmonary artery smooth muscle cells （PASMCs）. Methods Pulmonary artery smooth muscle cells were isolated from adult male SD rats and primarily cultured. The subcultured cells were harvested and divided into control group, hypoxia group, hypoxia group treated with FTY720 （5000ng/ml） and SB203580 （5p, mol/L） and hypoxia group treated with FTY720 （2500ng/ml,5000ng/ml）. The protein expression of P38MAPK , p - P38MAPK ,NF-κB P65 ,p - NF-κB were detected by Western blot. TUNEL was applied to de- tect the apoptosis of PASMCs. Results The apoptosis rate of PASMCs was markedly reduced in H group compared with N group（ P 〈 0. 05）. The expression of p- P38MAPK and p- NF-κB P65 were significantly increased and had no difference in the total P38MAPK and NF-κB P65 in H group. Compared with H group, the apoptosis rate of all treated groups was up regulated. The expression of p - P3gMAPK and p - NF-κB P65 were significantly decreased （ P 〈 0. 05 ,P 〈 0.01 ） in the manner of dose - dependent and had no differ- ence in the total P38MAPK and NF-κB P65 （ P 〉 0.05 ）. Conclusion FTY720 inhibits inflammatory response and proliferation of hy- poxic PASMCs by down regulation of the expression of p - P38MAPK and p - NF-κB and promotes apoptosis.
出处
《医学研究杂志》
2015年第6期56-59,共4页
Journal of Medical Research
基金
浙江省科技计划项目(2012C37092)
作者简介
林丽娜，电子邮箱：wzlinlina@163．com
