摘要
目的 :研究缺氧对体外培养的大鼠肺动脉平滑肌细胞诱导型血红素氧合酶 (HO - 1)基因表达及内源性一氧化碳 (CO)产生的影响 ,探讨HO -CO系统在缺氧性肺动脉高压中的作用。方法 :原代培养大鼠肺内动脉的平滑肌细胞 (PASMC)并传代 ,选用第 3- 5代PASMC用于实验。在培养瓶内通缺氧气体 (95 %N2 ,5 %CO2 ) 12、2 4h和48h后 ,用逆转录 -聚合酶链式反应 (RT -PCR)检测PASMC中HO - 1mRNA水平 ,用酶联免疫检测仪测定培养基中碳氧血红蛋白 (COHb)含量 ,放免法测定培养基中环磷酸鸟苷 (cGMP)含量。结果 :缺氧 12、2 4和 48hHO - 1mRNA分别较正常对照组高 2 7%、5 7%和 2 7 1% (P <0 0 1) ,培养基中COHb的含量分别较正常对照组高 13 8%、31 0 %和93 1% (P <0 0 1) ,PASMC中cGMP的含量分别为正常对照组的 2 7、4 0和 6 8倍 (P <0 0 1或P <0 0 5 )。结论 :缺氧诱导PASMC中的HO - 1基因表达上调 ,并使内源性CO产生增多 ,对PASMC内cGMP浓度有调节作用。
AIM: To investigate the effect of hypoxia on the expression of the inducible heme oxygenase (HO-1) gene and the production of endogenous carbon monoxide in pulmonary artery smooth muscle cells(PASMCs) METHODS: Primary culture of rat intra-pulmonary artery smooth muscle cells and the PASMCs were passed every 3 days, the 3-5 passages were used After exposure to hypoxic condition (95%N_2, 5%CO_2)12, 24 and 48 h, the level of HO-1 mRNA was examined by reverse transcriptase polymerase chain reaction (RT-PCR), the volume of COHb in the media were measured spectrophotometrically, the cGMP concentration of cell extracts was determined by radioimmunoassay RESULTS: After exposure to hypoxic condition for 12, 24, 48 h, the HO-1 mRNA increased by 2 7%,5 7% and 27 1%, respectively( P< 0 01); the HbCO in the media increased by 13 8%,31 0% and 93 1% ( P< 0 01); the cGMP concentration were 2 7, 4 0 and 6 8-folds as that of the control group ( P< 0 01 or P< 0 05) CONCLUSION: The expression of HO-1 gene in PASMCs is upregulated by hypoxia, the production of endogenous carbon monoxide is elevated as well
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2002年第1期86-88,共3页
Chinese Journal of Pathophysiology
