CAJ | 학술논문

目的检测大鼠肺微血管内皮细胞(PMVECs)内磷酸二酯酶(PDE)同功酶mRNA表达及基础酶活性,明确PMVECs存在的PDE亚型及体外活性. 方法采用植块法分离培养PMVECs,RT-PCR法测定PDE mRNA表达,以高效液相色谱(PHLC)检测环核苷酸在酶反应前后的含量变化,计算PDE活性. 结果检测结果显示,PMVECs中含有PDE1A、1C、2A、3A、3B、4A、4B、4C、4D、5A、7A、7B、8A、8B、9A、10A、11A共17种PDE mRNA,而PDE1B mRNA 未见表达,酶量在5～20μl范围内与PDE活性存在良好的线性关系. 结论 PMVECs中存在17种PDE基因表达,并有较高的基础酶活性.
목적 검측대서폐미혈관내피세포(PMVECs)내린산이지매(PDE)동공매mRNA표체급기출매활성,명학PMVECs존재적PDE아형급체외활성. 방법 채용식괴법분리배양PMVECs,RT-PCR법측정PDE mRNA표체,이고효액상색보(PHLC)검측배핵감산재매반응전후적함량변화,계산PDE활성. 결과 검측결과 현시,PMVECs중함유PDE1A、1C、2A、3A、3B、4A、4B、4C、4D、5A、7A、7B、8A、8B、9A、10A、11A공17충PDE mRNA,이PDE1B mRNA 미견표체,매량재5～20μl범위내여PDE활성존재량호적선성관계. 결론 PMVECs중존재17충PDE기인표체,병유교고적기출매활성.
Objective To study the main subtypes messenger ribonucleic acid(mRNA) and the basal enzyme activity of phosphodiesterase (PDE) in rat pulmonary microvascular endothelial cells (PMVECs) through the examination mRNA expression and activity of PDE in vitro. The data were offered to reveal the relationship between PDE distributions, activity change and to accumulate data for the possibility of drug regulation of its functional alteration. Methods The cells were cultured with tissue-sticking method;the gene expression of PDEs was detected by reverse transcript polymerase chain reaction (RT-PCR), and the activity of PDEs was calculated by cyclic nucleotides content change examined with high performance liquid chromatogram (HPLC) before and after the PDE reaction( n ＝3). Results The PMVECs identified by cell immunofluorescence with polyclonal antibody of CD31 were dissociated and cultured, mRNAs of PDE1A, 1C, 2A,3A, 3B, 4A, 4D, 5A, 7A, 7B, 8A, 8B, 9A, 10A,11A were expressed in PMVECs, but there was no mRNA of PDE1B expressed in PMVECs. cAMP/cGMP-PDE in the extent of 5-20μl had a good linear correlation with its activity. Conclusion There are 17 kinds of PDE gene expression existing in PMVECs which contain of the basic enzyme with a higher activity.