野百合碱对培养的肺动脉内皮细胞NO含量、eNOS蛋白表达及肺动脉平滑肌细胞收缩性的影响

目的研究野百合碱(monocrotaline pyrrole,MCTP)对培养的牛肺动脉内皮细胞(calf pulmonary artery endothelial cells,CPAEs)一氧化氮(NO)含量、内皮型一氧化氮合酶(eNOS)蛋白表达及肺动脉平滑肌细胞收缩的影响。方法采用荧光共聚焦激光显微镜检测培养的牛肺动脉内皮细胞的NO含量;Western blot检测eNOS蛋白表达;胶原凝胶实验分析肺动脉平滑肌细胞收缩性。结果经野百合碱处理的牛肺动脉内皮细胞NO含量和eNOS蛋白表达较对照组明显减少、肺动脉平滑肌细胞收缩性增强。结论MCTP可抑制eNOS蛋白表达,肺动脉内皮细胞NO产生受到抑制,使肺动脉平滑肌细胞收缩性增强,这些改变与肺动脉高压的形成可能存在一定关系。

Effects of MCTP on production of nitric oxide and expression of eNOS protein of cultured pulmonary artery endothelial cells and contractility of cultured pulmonary artery smooth muscle cells

Aim To investigate the effects of monocrotaline pyrrole on production of nitric oxide and expression of eNOS protein of cultured pulmonary artery endothelial cells and on contractility of cultured pulmonary artery smooth muscle cells.Methods DAF-2 fluorescence technique was used to determine NO level,Western blot analysis was performed to determine the level of eNOS protein,and collagen gel contraction system was adopted to analyze muscle contractility.Results NO production induced by ACh and expression of eNOS protein were obviously inhibited by monocrotaline pyrrole compared with those of control group and gel contraction area in MCTP-treated cells induced by Thapsigargin obviously decreased.Conclusions monocrotaline pyrrole could inhibit the level of the ACh-induced production of NO and expression of eNOS protein,and enhance the contractility of pulmonary artery smooth muscle cells,which may be one of the possible mechanisms of MCTP-induced pulmonary artery hypertension.