蛇床子素抗肺动脉高压作用及机制研究

Pulmonary arterial hypertension (PAH) is a refractory disease. Improvement of disorder of the proliferation and apoptosis in pulmonary artery smooth muscle cells (PASMCs) are the key of anti-PAH. Reinforcement of NO/cGMP/PKG signaling and inhibition of the nuclear translocation of NF-κB and NFAT are important pathways for anti-PAH. Combined the literature with our pre-experimental research results, it is speculated that the osthole (Ost) may posses anti-PAH effect, and the mechanisms may be due to modulating the above key aspects . Hence,the current study is firstly designed to study the effect of Ost on PAH induced by MCT in rats to identify anti-PAH effect and explore the possible mechanisms. Then using PDGF-BB stimulated PASMCs in vitro, the change of NO/cGMP signaling, intracellular [Ca2+]i, the nuclear translocation of NF-κB (p65/p50), NFATc2 and mitochondrial membrane potential as well as the expression of IL-2, IL-6, TNFα, KV1.5 in the PASMCs are detected in order to further analyze the mechanisms of improvement disorder of proliferation/apoptosis of PASMCs for Ost in PAH. The significance of the study is to provide the basic pharmacology data for Ost on anti-PAH and to quest for effective drug.

肺动脉高压（PAH）为难治性疾病，纠正肺动脉平滑肌（PASMCs）的增殖/凋亡平衡紊乱是其治疗的关键，增强NO/cGMP/PKG信号和抑制NF-κB及NFAT核转位是抗PAH的重要策略。本课题组结合文献及预实验研究推测蛇床子素 (Ost) 可能通过上述环节产生抗PAH效应，但缺乏系统的深入研究。因此，本项目首先观察Ost对野百合碱（MCT）所致的大鼠PAH模型的干预作用，确证其抗PAH效应并分析其可能机制；然后研究Ost作用于PDGF刺激的原代培养的PASMCs体系，观察Ost对NO/cGMP/PKG信号、细胞内游离钙、NF-κB（p65/p50）及NFATc2核转位的影响，以及转录产物IL-2、IL-6、TNF-α与KV1.5等表达及线粒体膜电位的变化，深入分析Ost 改善PASMCs增殖/凋亡紊乱的调控机制。本项目旨在为Ost用于PAH的治疗提供基础药理学依据，探索有效抗PAH药物。

肺动脉高压（PAH）为难治性疾病，纠正肺动脉平滑肌（PASMCs）的增殖/凋亡平衡紊乱是其治疗的关键，抑制NF-κB炎症通路是抗PAH的重要策略。本课题组结合文献及预实验研究推测中药蛇床子的有效成分蛇床子素(Ost)可能具有抗PAH的作用，但缺乏系统的研究。故本研究采用野百合碱（MCT）诱导的PAH和右心室重构模型，观察Ost对PAH和右心室重构的干预作用，并探讨其可能的机制。同时采用PDGF-BB刺激的肺动脉平滑肌细胞进一步研究Ost抗PAH的作用机制。研究发现：（1）Ost具有抗MCT所致大鼠PAH的作用，其可能的机制有：① Ost通过抑制PASMCs增殖、促进其凋亡减轻PAH；② Ost通过增强Nrf-2/ARE途径降低氧化应激水平抗PAH；③ Ost通过抑制NF-κB炎症通路减轻大鼠炎症反应；④ Ost通过下调TGF-β1/Smad2通路抗肺动脉重构。（2）Ost具有抗MCT所致大鼠右心室重构的作用，其可能的机制有：① Ost通过调控Bax/Bcl-2等的表达抑制心肌细胞凋亡；② Ost通过抑制NF-κB通路减轻大鼠炎症反应；③ Ost通过上调PPARα和PPARγ抗右心室重构。该研究表明中药蛇床子的有效成分Ost具有抗PAH和右心室重构的作用，该发现为Ost用于抗PAH和右心室重构的治疗提供了基础药理学依据，也为本省道地中药材的开发提供了相应的理论基础。