Zhang Hong-Liang, Liu Zhi-Hong, Luo Qin, Wang Yong, Zhao Zhi-Hui
State Key Laboratory of Cardiovascular Disease, Center for Pulmonary Vascular Diseases, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100037, People's R China.
Int J Clin Exp Med. 2015 Feb 15;8(2):1834-43. eCollection 2015.
Recent researches have shown that dysfunctional intracellular vesicular trafficking exists in pulmonary arterial hypertension (PAH). However, the expression of proteins involved in intracellular vesicular trafficking in pulmonary vasculature in PAH remains unclear.
To elucidate possible roles of proteins involved in intracellular vesicular trafficking in the development of PAH in rats treated with monocrotaline, changes in the expression of N-ethyl-maleimide-sensitive factor (NSF), α-soluble NSF attachment protein (α-SNAP) and synaptosome-associated membrane protein (SNAP) 23 were examined together with expression of caveolin-1 (cav-1), endogenous nitric oxide synthase (eNOS), type 2 bone morphogenetic receptor (BMPR2) and cellular apoptosis.
The mRNA expression was investigated by real time-PCR and protein expression by immunoblot method in rat lung. Caspase-3 was used as an indicator of cellular apoptosis and examined by immunoblot method.
During the development of PAH, mRNA and protein expression of NSF, α-SNAP and SNAP23 all significantly increased before pulmonary arterial pressure started to increase, then all significantly decreased when PAH established. The expression of eNOS and BMPR2 changed similarly, while the mRNA and protein of cav-1 both downregulated after monocrotaline treatment. Caspase-3 was also increased after exposure to monocrotaline.
Since the expression of NSF, α-SNAP and SNAP23 changed greatly during the onset of PAH and accompanied with abnormal expression of eNOS, BMPR2 and cav-1 and with enhanced cellular apoptosis, NSF, α-SNAP and SNAP23 appear to be associated with the development of PAH in rats treated with monocrotaline.
近期研究表明，肺动脉高压（PAH）中存在细胞内囊泡运输功能障碍。然而，PAH患者肺血管中参与细胞内囊泡运输的蛋白质表达仍不清楚。
为阐明参与细胞内囊泡运输的蛋白质在使用野百合碱处理的大鼠PAH发生发展中的可能作用，检测N - 乙基 - 马来酰亚胺敏感因子（NSF）、α - 可溶性NSF附着蛋白（α - SNAP）和突触体相关膜蛋白（SNAP）23的表达变化，同时检测小窝蛋白 - 1（cav - 1）、内源性一氧化氮合酶（eNOS）、2型骨形态发生蛋白受体（BMPR2）的表达及细胞凋亡情况。
采用实时定量PCR法检测大鼠肺组织中mRNA表达，免疫印迹法检测蛋白质表达。以半胱天冬酶 - 3作为细胞凋亡指标，采用免疫印迹法进行检测。
在PAH发展过程中，NSF、α - SNAP和SNAP23的mRNA和蛋白质表达在肺动脉压开始升高前均显著增加，而在PAH形成时均显著降低。eNOS和BMPR2的表达变化相似，而野百合碱处理后cav - 1的mRNA和蛋白质均下调。暴露于野百合碱后半胱天冬酶 - 3也增加。
由于NSF、α - SNAP和SNAP23的表达在PAH发病过程中变化显著，并伴有eNOS、BMPR2和cav - 1的异常表达以及细胞凋亡增加，NSF、α - SNAP和SNAP23似乎与用野百合碱处理的大鼠PAH的发生发展有关。
