【摘要】 目的:研究低氧内皮细胞条件培养液(HEC-CM)和DDPH对肺动脉平滑肌细胞(PASMC)骨桥蛋白(OPN)基因表达和α平滑肌肌动蛋白(α-SM-actin)的影响,进而探讨DDPH对增殖后PASMC的逆转效应。方法:利用HECCM建立猪PASMC的增殖模型;采用细胞培养、改良的四甲基偶氮唑盐比色法(MTT)、免疫细胞化学染色测定α-SM-actin、逆转录-聚合酶链反应(RT-PCR)检测OPN基因表达等方法观察HECCM的促PASMC增殖作用,以及DDPH对增殖后PASMCα-SM-actin和OPN表达的影响。结果:(1)HECCM促PASMC增殖后,PASMC的α-SM-actin含量降低而其OPN基因表达上调;(2)HECCM促PASMC增殖后,加入DDPH可使PASMC的α-SM-actin含量回升,OPN表达下调。结论:DDPH对HECCM促PASMC增殖有逆转效应,可使增殖后PASMC由合成表型逆转为正常收缩表型。更多还原

【Abstract】 AIM:To study the effects of hypoxial endothelia cell conditional medium(HECCM) and DDPH on the expression of osteopontin(OPN) gene and α-SM-actin in pulmonary arterial muscle cell(PASMC),and the reverse effect of DDPH on proliferating PASMC induced by HECCM.METHODS:Hypoxial endothelia cell model was established in PASMC of swine.MTT assay and Immunocytochemistry was used to test cell proliferation,the expression of α-SM-actin.Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of OPN gene.RESULTS:(1)HECCM decrease the level of α-SM-actin and up-regulate the expression of OPN gene in PASMC;(2)DDPH increase the level of α-SM-actin and down-regulate the expression of OPN gene in PASMC.CONCLUSION:DDPH could reverse the phenotype transformation of PASMC exposed to HECCM,and transform PASMC from synthetic phenotype to normal contractile phenotype.更多还原