猪松弛素对人肺微血管内皮细胞产生一氧化氮的影响

Effect of porcine relaxin on NO production of human microvascular endothelial cells

目的观察猪松弛素(pRLX)对人肺微血管内皮细胞(HMVECs)产生一氧化氮(NO)的影响,探讨这种效应的可能机制。方法Western blotting检测不同浓度pRLX作用下HMVECs诱生型一氧化氮合成酶(iNOS)和原生型一氧化氮合成酶(cNOS)蛋白表达;Griess法检测不同浓度pRLX分别在不同的作用时间下对HMVECs NO产量的影响;Griess法检测选择性iNOS抑制剂氨基胍、非选择性NOS抑制剂L-单甲基精氨酸和NF-κB抑制剂吡咯烷二硫氨基甲酸对pRLX刺激HMVECs产生NO效应的影响。结果pRLX促进HMVECs iNOS蛋白表达,但对cNOS表达影响不显著;pRLX呈浓度依赖性促进HMVECs NO产量,但24 h至96 h范围内各pRLX作用时间组之间NO产量无显著差异;氨基胍、L-单甲基精氨酸和吡咯烷二硫氨基甲酸均能阻断pRLX促进HMVECs产生NO的效应。结论pRLX促进HMVECs iNOS表达和NO生成。

Objective To observe effect of porcine relaxin（pRLX） on NO production of human microvascular endothelial cells （HMVECs） and discuss its possible mechanism. Methods iNOS and cNOS expression of HMVECs with or without pRLX were detected using western blotting. NO production of HMVECs with pRLX at different concentration or different time were determined by method of Griess. NO production of pRLX of HMVECs plus Non-selective NOS inhibitor NG-monomethyl-L-arginine （L-NMMA）, selective iNOS inhibitor aminoguanidine （AG） or nuclear factors-κ B （NF-κ B） inhibitor pyrrolidine dithiocarbamate （PDTC） were also analysed. Results pRLX promoted iNOS protein expression of HMVECs, but not cNOS protein expression. NO production of HMVECs was promoted by pRLX on concentration-dependent pattern instead of time-dependent one. AG, L-NMMA and PDTC were showed to block the effect of pRLX on NO production of HMVECs. Conclusion pRLX promote iNOS expression and NO production of HMVECs.