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Carbon monoxide inhibits proliferation of pulmonary smooth muscle cells under hypoxia 被引量:2
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作者 甄国华 张珍祥 +1 位作者 薛峥 徐永健 《中华医学杂志:英文版》 SCIE CAS CSCD 2003年第12期 1804-1809,共6页
Objective To investigate the expression of inducible heme oxygenase (HO-1) gene in pulmonary artery smooth muscle cells (PASMCs) exposed to hypoxia, and the influence of carbon monoxide (CO) on the proliferation of PA... Objective To investigate the expression of inducible heme oxygenase (HO-1) gene in pulmonary artery smooth muscle cells (PASMCs) exposed to hypoxia, and the influence of carbon monoxide (CO) on the proliferation of PASMCs under hypoxic conditions.Methods Primary culture of rat PASMCs were passed every 3 days, and the 3-5 passages were used. After exposure to hypoxic conditions (95% N2, 5% CO2) 0, 12, 24 and 48 hours, the level of HO-1 mRNA was examined by reverse transcriptase polymerase chain reaction (RT-PCR). The volume of COHb in the medium was measured spectrophotometrically. The cyclic guanosine mono-phosphate (cGMP) concentration of cell extracts was determined by radioimmunoassay. PASMCs were divided into 5 groups, cultured under normoxia and hypoxia and treated with hemin, hemoglobin (Hb) and exogenous CO respectively. Then 3-(4, 5-cimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) colorimetric assay and immunocytochemical staining were used to study the energy metabolism and the expression of proliferating cell nuclear antigen (PCNA) in PASMCs. Flow cytometry was used to analyze the cell cycle of PASMCs.Results After exposure to hypoxic conditions for 12, 24, and 48 hours, the HO-1 mRNA increased by 2.7%, 5.7% and 27.1% respectively (P<0.01). The carboxy-hemoglobin (COHb) in the medium increased by 13.8%, 31.0% and 93.1% (P<0.01); the cGMP concentrations were 2.7, 4.0 and 6.8-fold compared with the control group (P<0.01 and P<0.05). In comparison with the control group, the value of MTT colorimetric assay, the immunocytochemical staining of PCNA and the percentages of PASMCs in S and G2M phases in the hypoxic group were significantly higher (P<0.01). After treatment with Hemin and CO, the results of the above analysis decreased significantly (P<0.01 and P<0.05), but increased significantly after treatment with Hb (P<0.01 and P<0.05).Conclusions The expression of HO-1 gene in PASMCs is upregulated by hypoxia and the production of endogenous CO is elevated as well. The endogenous CO suppresses 展开更多
关键词 肺动脉平滑肌细胞 细胞增殖 一氧化碳 组织缺氧 亚铁血红素氧酶 RT-PCR
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