摘要
目的观察油酸肺损伤早期肺组织一氧化氮合成酶（ＮＯＳ）活性变化、诱导型一氧化氮合成酶（ｉＮＯＳ）基因表达变化及其细胞定位。方法３Ｈ－胍氨酸测定法测定ＮＯＳ活性；应用３２Ｐ标记核酸探针狭线杂交技术和地高辛标记核酸探针原位杂交技术检测油酸肺损伤大鼠肺组织。结果正常肺组织ＮＯＳ活性很低且无ｉＮＯＳｍＲＮＡ表达；急性肺损伤后ｉＮＯＳ活性升高且与ｉＮＯＳｍＲＮＡ表达相一致；硝基左旋精氨酸（ＬＮＮＡ）可抑制ｉＮＯＳ活性并下调ｉＮＯＳｍＲＮＡ表达；伤后表达ｉＮＯＳｍＲＮＡ的阳性细胞可能为肺泡巨噬细胞、粒细胞、支气管上皮细胞、肺泡上皮细胞、平滑肌细胞、内皮细胞。结论急性肺损伤后肺组织ｉＮＯＳｍＲＮＡ的表达是一个突然启动的主动合成过程；ＬＮＮＡ抑制ＮＯ产生可能与其下调ｉＮＯＳｍＲＮＡ表达有关；肺组织表达ｉＮＯＳｍＲＮＡ阳性细胞的多样性可能与ＮＯ损伤肺组织的复杂性有关。
Aim To investigate the biological activity and gene expression of nitric oxide synzyme (NOS) and its cell location in lung tissue after acute lung injury. Methods 3H L argine was used to measure the NOS activity by dot blot hybridization labled with 32 P cDNA probe and in situ hybridization labled with digoxin cDNA probe was used to detect the gene expression. Results It was found that although there was low level activity of NOS and no iNOS mRNA transcription in the lung tissue under normal condition, postinjury induced inflammatory mediators could initiate iNOS mRNA transcription in the pulmonary tissue, it reached the peak value at 2 hours after injury then went down steadily, and the change of iNOS activity was similar to that of iNOS mRNA transcription. N nitro L arginine (LNNA) could inhibit the activity of NOS and down regulate the transcription of iNOS mRNA. The cells expressing iNOS mRNA were alveolar macrophages, granuocytes, bronchial and alveolar epithelial cells, smooth muscle cells and endothelial cells respectively. Conclusion The transcription of iNOS mRNA in lung tissue after pulmonary injury was characterized to be a sudden started and actively compound course and the inhibition of NO production by LNNA might be related to its downregulation of iNOS mRNA transcription. So many kinds of cells which express iNOS gene reflect the complicated mechanism of acute lung injury.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
1998年第6期374-376,共3页
Chinese Journal of Trauma
关键词
肺损伤
一氧化氮合成酶
原位杂交
基因表达
Acute lung injury Nitric oxide synzyme Dot blot hybridization In situ hybridization
