清热中药对大鼠肺及其微血管内皮细胞磷酸二酯酶活性的影响

The Effect Heat-clearing Herbs to the Activity of Phosphodiesterase in Rat Lung and Pulmonary Microvascular Endothelial Cells

采用组织贴块法培养分离大鼠肺微血管内皮细胞（PMVEC），以高效液相色谱（HPLC）检测环核苷酸（cAMP）在酶反应前后磷酸二酯酶（PDE）的变化计算PDE活性，并分析药物对其影响，为研究PMVEC特异性PDE抑制剂和清热药作用机理积累资料。结果显示：分离培养经CD31鉴定的PMVEC，其酶量在5-20μL范围内与PDE活性存在良好的线性关系（r=0.9929）；17味入肺经清热药均对肺组织cAMP-PDE酶活性有不同程度的抑制作用，其中黄芩、黄药子、青蒿抑制率较高，对肺组织酶分别为：75.37%，72%，61.26%，对PMVEC分别为：66.1%，63.54%，59.39%，结果提示以cAMP-PDE活性为指标，有望筛选出其特异性PDE抑制剂，并揭示入肺经清热药的作用机制。

Rat pulmonary microvascular endothelial cells （PMVEC） were dissociated and cultured with tissue-sticking method; The activity of phosphodiesterase（PDE） was calculated by the content changes of cyclic nucleotides before and after the PDE reaction examined by high performance liquid chromatography （HPLC） to analyze the effects of heat-clearing herbs to cAMP-PDE activity. The results showed that： The PMVEC identified by cell immunostainin with CD31 were dissociated and cultured; cAMP-PDE in the extent of 5-20μL had a good linear correlation with its activity （r=0. 9929） ; The activity of cAMP-PDE in rat lung and PMVEC were inhibited differently by all the seventeen heat-clearing drugs, in which the inhibition to lung of Huangqin, Huangyaozi, Qinghao were 75.37%, 72%, 61.26%, to PMVEC were 66.1%, 63.54%,59.39%; The data offered above could be used to choose out the inhibition of PDE through regarding cAMP-PDE as a kind of indicatrix and reveal the mechanism of heat-clearing drugs going to the lung meridian.