P268 The role of growth and differentiation factor 15 in smooth muscle cell proliferation in pulmonary hypertension
P268 The role of growth and differentiation factor 15 in smooth muscle cell proliferation in pulmonary hypertension
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P268 生长分化因子15在肺动脉高压平滑肌细胞增殖中的作用
DOI：
10.1136/thoraxjnl-2015-207770.404
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发表时间：
2015
期刊：
影响因子：
10
通讯作者：
Garfield B
中科院分区：
文献类型：
--
作者：
Garfield B
生长分化因子15（GDF-15）是肺动脉高压（PH）的预后指标。其对内皮细胞的影响已被记录，但其作用机制和在PH发展中的作用尚未得到充分研究。我们的目的是定义的作用和GDF-15的作用机制，在PH.MethodsRats的发展与moncrotaline（MCT）或车辆控制和安乐死后，进行心血管监测4周后。通过qPCR测量肺中GDF-15 mRNA的表达。通过ELISA分析血清和肺中的总GDF-15蛋白水平。用免疫组化法分析GDF-15在肺组织中的分布。GDF-15信号在人肺动脉平滑肌细胞（HPASMCs）进行了分析，使用western blot，和它的作用进行了测量，用cyquant assay.ResultsGDF-15 mRNA和蛋白水平的增加，在肺匀浆中的MCT大鼠相比，对照组（p < 0.05）。免疫组化显示GDF-15定位于内皮细胞，在这些动物的PASMC中的程度较低。MCT给药大鼠血清中的GDF-15水平高于溶剂对照组（771 ± 345 vs. 411 ± 305，p < 0.05）。在MCT治疗组中，血清GDF-15与RV/LV+S重量相关（Pearson r = 0.66，p < 0.05）。免疫组化还显示MCT大鼠PASMC中磷酸化TGF β激活激酶1（TAK 1）增加。在HPASMCs中，GDF-15（1 ng/ml）处理导致72 h时增殖相对于基线增加（图1）。GDF-15也能够诱导磷酸化的TAK 1在HPASMCs.ConclusionsGDF-15是过度表达在肺血管的MCT大鼠，模仿人类疾病。GDF-15与这些动物的右心室肥大程度相关。GDF-15下游信号分子磷酸化TAK-1在MCT大鼠血管中的水平增加。体外GDF-15处理引起HPASMC增殖和TAK-1活化。需要对该途径进行进一步研究以确定其与人类疾病的相关性。
IntroductionGrowth and differentiation factor 15 (GDF-15) is a prognostic marker in pulmonary hypertension (PH). Its effects on endothelial cells have been documented, but its mechanism of action and role in the development of PH have not yet been fully investigated. We aimed to define the role and mechanism of action of GDF-15 in the development of PH.MethodsRats were treated with moncrotaline (MCT) or vehicle control and euthanized after undergoing cardiovascular monitoring 4 weeks later. The expression of GDF-15 mRNA in the lung was measured by qPCR. Total GDF-15 protein levels in serum and lung were analysed by ELISA. The distribution of GDF-15 in the lung was analysed by immunohistochemistry. GDF-15 signalling in human pulmonary artery smooth muscle cells (HPASMCs) was analysed using western blot, and its role on HPASMC proliferation was measured using a cyquant assay.ResultsGDF-15 mRNA and protein levels were raised in the lung homogenates of the MCT rat compared to controls (p < 0.05). Immunohistochemistry revealed GDF-15 was localised in the endothelial cells and to a lesser extent in the PASMCs of these animal. GDF-15 levels in the serum of the MCT treated rats was higher than that in those treated with vehicle control (771 ± 345 vs. 411 ± 305, p < 0.05). Serum GDF-15 was correlated with RV/LV+S weight in the MCT treated group (Pearson r = 0.66, p < 0.05). Immunohistochemistry also revealed an increase of phospho-TGFβ activated kinase 1 (TAK1) in PASMCs of the MCT rat. In HPASMCs GDF-15 (1 ng/ml) treatment resulted in an increase in proliferation over baseline at 72 h (Figure 1). GDF-15 was also able to induce phosphorylation of TAK1 in HPASMCs.ConclusionsGDF-15 is over-expressed in the lung vasculature of MCT rats, mimicking human disease. GDF-15 was associated with the degree of right ventricular hypertrophy in these animals. GDF-15 downstream signalling molecule phosphorylated TAK-1 is present in increased levels in the vasculature of the MCT rat.In vitroGDF-15 treatment caused proliferation of HPASMCs and activation of TAK-1. Further investigation of this pathway is required to determine its relevance to human disease.