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本研究应用细胞培养、3H-TdR掺入、核酸分子杂交、免疫组织化学染色技术,探讨无氧(0%O2+95%N2+5%CO2)和/或低氧(2.5~3%O2+92%N2+5%CO2)对新生小牛肺动脉平滑肌细胞(PASM)增殖和5-羟色胺转载体基因表达的影响。结果表明:无氧24h可刺激PASM的DNA合成,3H-TdR的掺入增加(P<0.05),加入5-羟色胺能非常显著地促进无氧PASM增殖(P<0.001),而对常氧PASM无明显促增殖作用,无氧5-羟色胺组与单纯无氧组相比,3H-TdR掺入值增加也非常显著(P<0.01);免疫组织化学染色分析证实无氧组PASM5-羟色胺含量较常氧组明显减少(P<0.001),分子杂交显示无氧组和低氧组5-羟色胺转载体mRNA表达较常氧组显著增加(PM<O.01)。结果提示:缺氧时5-羟色胺促进PASM增殖,5-羟色胺转载体基因表达增加,可能是缺氧肺动脉高压形成的机制之一。
In this study, we investigated the effects of anaerobic (0% O2 + 95% N2 + 5% CO2) and / or hypoxia (2.5-3% O2 + 92% N2 + 5%) on cell proliferation, 3H-TdR incorporation, hybridization of nucleic acid molecules and immunohistochemical staining. CO2) on proliferation and serotonin gene expression of newborn calf pulmonary artery smooth muscle cells (PASM). The results showed that anaerobic 24 h stimulation of DNA synthesis of PASM resulted in an increase of 3H-TdR incorporation (P <0.05). Addition of 5-HT could significantly promote the proliferation of anaerobic PASM (P <0.001) There was no significant effect on the proliferation of normoxia PASM. The 3H-TdR incorporation was also significantly increased in the anaerobic 5-HT group compared with the anaerobic group (P <0.01). Immunohistochemical staining confirmed that there was no oxygen Compared with normoxic group, PASM serotonin decreased significantly (P <0.001). Molecular hybridization showed that mRNA expression of serotonin transcripts increased significantly in hypoxia group and hypoxia group compared with normoxia group (P <0.001). The results suggest that serotonin can promote the proliferation of PASM and increase the gene expression of serotonin in hypoxia, which may be one of the mechanisms of hypoxic pulmonary hypertension.