Stable isotope metabolomics of pulmonary artery smooth muscle and endothelial cells in pulmonary hypertension and with TGF-beta treatment.,Scientific Reports

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Stable isotope metabolomics of pulmonary artery smooth muscle and endothelial cells in pulmonary hypertension and with TGF-beta treatment.
Scientific Reports ( IF 3.8 ) Pub Date : 2020-01-15 , DOI: 10.1038/s41598-019-57200-5
Daniel Hernandez-Saavedra ₁ , Linda Sanders ₁ , Scott Freeman ₁ , Julie A Reisz ₂ , Michael H Lee ₁ , Claudia Mickael ₁ , Rahul Kumar _{1,

3} , Biruk Kassa _{1,

3} , Sue Gu ₁ , Angelo D' Alessandro ₂ , Kurt R Stenmark ₄ , Rubin M Tuder ₁ , Brian B Graham _{1,

3}

Affiliation

Altered metabolism in pulmonary artery smooth muscle cells (PASMCs) and endothelial cells (PAECs) contributes to the pathology of pulmonary hypertension (PH), but changes in substrate uptake and how substrates are utilized have not been fully characterized. We hypothesized stable isotope metabolomics would identify increased glucose, glutamine and fatty acid uptake and utilization in human PASMCs and PAECs from PH versus control specimens, and that TGF-β treatment would phenocopy these metabolic changes. We used 13C-labeled glucose, glutamine or a long-chain fatty acid mixture added to cell culture media, and mass spectrometry-based metabolomics to detect and quantify 13C-labeled metabolites. We found PH PASMCs had increased glucose uptake and utilization by glycolysis and the pentose shunt, but no changes in glutamine or fatty acid uptake or utilization. Diseased PAECs had increased proximate glycolysis pathway intermediates, less pentose shunt flux, increased anaplerosis from glutamine, and decreased fatty acid β-oxidation. TGF-β treatment increased glycolysis in PASMCs, but did not recapitulate the PAEC disease phenotype. In TGF-β-treated PASMCs, glucose, glutamine and fatty acids all contributed carbons to the TCA cycle. In conclusion, PASMCs and PAECs collected from PH subjects have significant changes in metabolite uptake and utilization, partially recapitulated by TGF-β treatment.

中文翻译：

肺动脉高压和TGF-β治疗的肺动脉平滑肌和内皮细胞的稳定同位素代谢组学。

肺动脉平滑肌细胞（PASMCs）和内皮细胞（PAECs）代谢的改变有助于肺动脉高压（PH）的病理，但是底物摄取的变化以及底物的利用方法尚未完全阐明。我们假设稳定的同位素代谢组学将确定人PASMC和PAEC中PH和对照标本中葡萄糖，谷氨酰胺和脂肪酸的摄取和利用增加，并且TGF-β处理将表型化这些代谢变化。我们使用添加到细胞培养基中的13C标记的葡萄糖，谷氨酰胺或长链脂肪酸混合物，以及基于质谱的代谢组学来检测和定量13C标记的代谢物。我们发现PH PASMCs通过糖酵解和戊糖分流增加了葡萄糖的吸收和利用，但谷氨酰胺或脂肪酸的摄取或利用没有变化。患病的PAECs的糖酵解途径中间产物增加，戊糖分流减少，谷氨酰胺引起的动脉粥样硬化增加，脂肪酸β-氧化减少。TGF-β处理增加了PASMCs的糖酵解，但没有概括PAEC疾病的表型。在经过TGF-β处理的PASMC中，葡萄糖，谷氨酰胺和脂肪酸都为TCA循环贡献了碳。总之，从PH受试者收集的PASMC和PAEC在代谢物的吸收和利用方面有显着变化，部分通过TGF-β处理得以概括。但没有概括PAEC疾病的表型。在经过TGF-β处理的PASMC中，葡萄糖，谷氨酰胺和脂肪酸都为TCA循环贡献了碳。总之，从PH受试者收集的PASMC和PAEC在代谢物的吸收和利用方面有显着变化，部分通过TGF-β处理得以概括。但没有概括PAEC疾病的表型。在经过TGF-β处理的PASMC中，葡萄糖，谷氨酰胺和脂肪酸都为TCA循环贡献了碳。总之，从PH受试者收集的PASMCs和PAECs在代谢物吸收和利用方面有显着变化，部分通过TGF-β处理得以概括。

更新日期：2020-01-15

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