摘要
目的 :观察低浓度一氧化碳 (CO)和低氧处理肺动脉平滑肌细胞 (PASMC )后 ,血红素氧合酶 (HO)基因表达的状况。方法 :培养大鼠PASMC成功后并进行鉴定 ,应用免疫组织化学、逆转录聚合酶链式反应 (RT -PCR)和Westernblot的方法研究低浓度CO和低氧作用后PASMC的HOmRNA及HO蛋白质含量。结果 :①HO - 1抗体免疫组织化学染色低氧组细胞胞浆呈黄色 ,中等强度 ;CO处理组的细胞胞浆呈棕黄色 ,强阳性 ,比低氧组的颜色显著深。HO - 2抗体染色各组细胞胞浆均呈淡黄色 ,弱阳性。②低氧组细胞HO - 1mRNA的表达水平为 1.2 5± 0 .37,明显高于常氧组细胞 (0 .12± 0 .0 4 ) ;低浓度CO组细胞HO - 1mRNA的表达水平为 3.5 2± 0 .6 8显著高于低氧组。各组细胞HO - 2mRNA的水平无明显差异。③低氧 4 8h组细胞HO - 1的蛋白含量为 1.0 7± 0 .15 ,高于低氧 2 4h组细胞(0 .5 2± 0 .0 4 ) ;低浓度CO组细胞HO - 1的蛋白含量为 3.6 5± 0 .4 3,显著高于低氧组细胞 ,4 8h蛋白质含量为最高。结论 :低浓度CO和低氧处理PASMC ,可以促进HO - 1mRNA和蛋白质的表达 ,而HO - 1在细胞低氧损害中发挥调节作用 。
AIM: To study the expression level of heme oxygenase -1 and -2 in pulmonary arterial smooth muscle cell(PASMC) treated with carbon monoxide(CO). METHODS: (1) The expression of HO in cultured rat PASMC was determined by immunohistochemistry stain. (2) The levels of HO mRNA and protein were evaluated by reverse transcription- polymerase chain reaction(RT-PCR) and Western blot. RESULTS: (1) PASMC cytoplasm showed yellow (++-+++) in hypoxia group, the buffy color(++++) in CO group as cells were stained by HO-1 antibody. (2) The expression level of HO-1 mRNA (1.25±0.37) in hypoxia group was higher than that of normal group (0.12±0.04). It was much higher (3.52±0.68) in CO group, than that of hypoxia group. (3)The content of HO-1 protein (1.07±0.15) in hypoxia 48 h group was higher than that of hypoxia 24 h group(0.52±0.04). It was higher(3.65±0.43) in CO group than that of hypoxia group.It was highest in CO 48 h group. CONCLUSION: Low density of CO and hypoxia enhanced the expression of HO-1 mRNA and protein in PASMC when cell were treated in hypoxia or CO,indicating that HO-1 has regulatory effect on PASMC during hypoxia.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2002年第7期749-752,共4页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目 (No .39970 32 9)
