SphK1/S1P Mediates PDGF-Induced Pulmonary Arterial Smooth Muscle Cell Proliferation via miR-21/BMPRII/Id1 Signaling Pathway
SphK1/S1P Mediates PDGF-Induced Pulmonary Arterial Smooth Muscle Cell Proliferation via miR-21/BMPRII/Id1 Signaling Pathway
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DOI：
10.1159/000495243
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Li, Manxiang
中科院分区：
文献类型：
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作者：
Li, Fangwei;Wang, Jian;Li, Manxiang
背景/目标：鞘氨醇激酶1（SphK 1）/1-磷酸鞘氨醇（S1 P）介导血小板源性生长因子（PDGF）诱导的肺动脉平滑肌细胞（PASMC）增殖的潜在分子机制尚不清楚，本研究旨在解决这一问题。研究方法：采用小干扰RNA（siRNA）和microRNA抑制剂转染，阻断SphK 1、骨形态发生蛋白受体II（BMPRII）和microRNA-21（miR-21）的表达。免疫印迹法检测SphK 1、BMPRII和DNA结合抑制因子1（Id 1）的基因表达水平，qRT-PCR法检测miR-21的表达水平，ELISA法检测S1 P的产生。BrdU掺入法检测PASMC增殖。结果如下：结果表明，PDGF可促进PASMCs增殖，上调miR-21的表达，下调BMPRII和Id 1的表达，增加SphK 1蛋白的表达和S1 P的生成。用siRNA预沉默SphK 1逆转PDGF诱导的S1 P产生、miR-21上调、BMPRII和Id 1下调以及PASMC增殖。miR-21的预抑制也阻断了PDGF引起的BMPRII和Id 1下调以及PASMC增殖。BMPRII的敲低下调了PASMCs中Id 1的表达。我们进一步发现，抑制PI 3 K/Akt和ERK信号通路，特别是ERK级联，可抑制PDGF诱导的上述变化。结论：本研究提示SphK 1/S1 P通路通过miR-21/BMPRII/Id 1轴在PDGF诱导的PASMC增殖中发挥重要作用，靶向SphK 1/S1 P轴可能成为防治肺动脉高压（PAH）的新策略。
Background/Aims: The underlying molecular mechanisms involved in sphingosine kinase 1 (SphK1)/sphingosine 1-phosphate (S1P) mediation of platelet-derived growth factor (PDGF)-induced pulmonary arterial smooth muscle cell (PASMC) proliferation are still unclear, and the present study aims to address this issue. Methods: Small interfering RNA (siRNA) and microRNA inhibitor transfection was performed to block the expression of SphK1, bone morphogenetic protein receptor II (BMPRII) and microRNA-21 (miR-21). Gene expression levels of SphK1, BMPRII and inhibitor of DNA binding 1 (Id1) were detected by immunoblotting, miR-21 expression level was examined with qRT-PCR, and S1P production was measured by ELISA. Additionally, PASMC proliferation was determined by BrdU incorporation assay. Results: Our results indicated that PDGF increased the expression of SphK1 protein and S1P production, up-regulated miR-21 expression, reduced BMPRII and Id1 expression, and promoted PASMCs proliferation. Pre-silencing of SphK1 with siRNA reversed PDGF-induced S1P production, miR-21 up-regulation, BMPRII and Id1 down-regulation, as well as PASMC proliferation. Pre-inhibition of miR-21 also blocked BMPRII and Id1 down-regulation as well as PASMC proliferation caused by PDGF. Knockdown of BMPRII down-regulated Id1 expression in PASMCs. We further found that inhibition of PI3K/Akt and ERK signaling pathways, particularly ERK cascade, suppressed PDGF-induced above changes. Conclusion: Our study indicates that SphK1/S1P pathway plays an important role in PDGF-induced PASMC proliferation via miR-21/BMPRII/Id1 axis and targeting against SphK1/S1P axis might be a novel strategy in the prevention and treatment of pulmonary arterial hypertension (PAH).