Alamri Abdulwahab, Burzangi Abdulhadi S, Coats Paul, Watson David G
Department of Pharmacology, College of Pharmacy Sciences, University of Hail, 55476 Hail, Saudi Arabia.
Strathclyde Institute of Pharmacy and Biomedical Science, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK.
Metabolites. 2018 Nov 30;8(4):87. doi: 10.3390/metabo8040087.
Pulmonary arterial hypertension (PAH) is a multi-factorial disease characterized by the hyperproliferation of pulmonary artery smooth muscle cells (PASMCs). Excessive reactive oxygen species (ROS) formation resulted in alterations of the structure and function of pulmonary arterial walls, leading to right ventricular failure and death. Diabetes mellitus has not yet been implicated in pulmonary hypertension. However, recently, variable studies have shown that diabetes is correlated with pulmonary hypertension pathobiology, which could participate in the modification of pulmonary artery muscles. The metabolomic changes in PASMCs were studied in response to 25 mM of D-glucose (high glucose, or HG) in order to establish a diabetic-like condition in an in vitro setting, and compared to five mM of D-glucose (normal glucose, or LG). The effect of co-culturing these cells with an ideal blood serum concentration of cholecalciferol-D3 and tocopherol was also examined. The current study aimed to examine the role of hyperglycemia in pulmonary arterial hypertension by the quantification and detection of the metabolomic alteration of smooth muscle cells in high-glucose conditions. Untargeted metabolomics was carried out using hydrophilic interaction liquid chromatography and high-resolution mass spectrometry. Cell proliferation was assessed by cell viability and the [³H] thymidine incorporation assay, and the redox state within the cells was examined by measuring reactive oxygen species (ROS) generation. The results demonstrated that PASMCs in high glucose (HG) grew, proliferated faster, and generated higher levels of superoxide anion (O₂·) and hydrogen peroxide (H₂O₂). The metabolomics of cells cultured in HG showed that the carbohydrate pathway, especially that of the upper glycolytic pathway metabolites, was influenced by the activation of the oxidation pathway: the pentose phosphate pathway (PPP). The amount of amino acids such as aspartate and glutathione reduced via HG, while glutathione disulfide, N6-Acetyl-L-lysine, glutamate, and 5-aminopentanoate increased. Lipids either as fatty acids or glycerophospholipids were downregulated in most of the metabolites, with the exception of docosatetraenoic acid and PG (16:0/16:1(9Z)). Purine and pyrimidine were influenced by hyperglycaemia following PPP oxidation. The results in addition showed that cells exposed to 25 mM of glucose were oxidatively stressed comparing to those cultured in five mM of glucose. Cholecalciferol (D3, or vitamin D) and tocopherol (vitamin E) were shown to restore the redox status of many metabolic pathways.
肺动脉高压（PAH）是一种多因素疾病，其特征为肺动脉平滑肌细胞（PASMCs）过度增殖。过量活性氧（ROS）的形成导致肺动脉壁结构和功能改变，进而引发右心室衰竭和死亡。糖尿病与肺动脉高压尚无关联。然而，最近多项研究表明，糖尿病与肺动脉高压病理生物学相关，可能参与肺动脉平滑肌的改变。为了在体外建立类似糖尿病的状态，研究了PASMCs在25 mM D-葡萄糖（高糖，HG）作用下的代谢组学变化，并与5 mM D-葡萄糖（正常葡萄糖，LG）进行比较。还检测了将这些细胞与理想血清浓度的胆钙化醇-D3和生育酚共培养的效果。本研究旨在通过定量和检测高糖条件下平滑肌细胞代谢组学改变，来探究高血糖在肺动脉高压中的作用。采用亲水相互作用液相色谱和高分辨率质谱进行非靶向代谢组学分析。通过细胞活力和[³H]胸苷掺入试验评估细胞增殖，并通过测量活性氧（ROS）生成来检测细胞内的氧化还原状态。结果表明，高糖（HG）环境中的PASMCs生长、增殖更快，超氧阴离子（O₂·）和过氧化氢（H₂O₂）生成水平更高。在HG中培养的细胞代谢组学显示，碳水化合物途径，尤其是糖酵解上游途径代谢物，受到氧化途径（磷酸戊糖途径，PPP）激活的影响：天冬氨酸和谷胱甘肽等氨基酸的量通过HG减少，而谷胱甘肽二硫化物、N6-乙酰-L-赖氨酸、谷氨酸和5-氨基戊酸增加。除二十二碳四烯酸和PG（16:0/16:1(9Z)）外，大多数代谢物中的脂肪酸或甘油磷脂形式的脂质下调。嘌呤和嘧啶在PPP氧化后受到高血糖影响。结果还表明，与在5 mM葡萄糖中培养的细胞相比，暴露于25 mM葡萄糖的细胞受到氧化应激。胆钙化醇（D3，或维生素D）和生育酚（维生素E）显示可恢复许多代谢途径的氧化还原状态。
