Institute of Cell Biophysics RAS, Pushchino, Russia.
Moscow Institute of Physics and Technology (State University), Dolgoprudny, Russia.
Mol Biol Rep. 2019 Jun;46(3):3101-3112. doi: 10.1007/s11033-019-04765-z. Epub 2019 Apr 12.
Chronic overuse of common pharmaceuticals, e.g. acetaminophen (paracetamol), often leads to the development of acute liver failure (ALF). This study aimed to elucidate the effect of cultured mesenchymal stem cells (MSCs) proteome on the onset of liver damage and regeneration dynamics in animals with ALF induced by acetaminophen, to test the liver protective efficacy of MSCs proteome depending on the oxygen tension in cell culture, and to blueprint protein components responsible for the effect. Protein compositions prepared from MSCs cultured in mild hypoxic (5% and 10% O and normal (21% O) conditions were used to treat ALF induced in mice by injection of acetaminophen. To test the effect of reduced oxygen tension in cell culture on resulting MSCs proteome content we applied a combination of high performance liquid chromatography and mass-spectrometry (LC-MS/MS) for the identification of proteins in lysates of MSCs cultured at different O levels. The treatment of acetaminophen-administered animals with proteins released from cultured MSCs resulted in the inhibition of inflammatory reactions in damaged liver; the area of hepatocyte necrosis being reduced in the first 24 h. Compositions obtained from MSCs cultured at lower O level were shown to be more potent than a composition prepared from normoxic cells. A comparative characterization of protein pattern and identification of individual components done by a cytokine assay and proteomics analysis of protein compositions revealed that even moderate hypoxia produces discrete changes in the expression of various subsets of proteins responsible for intracellular respiration and cell signaling. The application of proteins prepared from MSCs grown in vitro at reduced oxygen tension significantly accelerates healing process in damaged liver tissue. The proteomics data obtained for different preparations offer new information about the potential candidates in the MSCs protein repertoire sensitive to oxygen tension in culture medium, which can be involved in the generalized mechanisms the cells use to respond to acute liver failure.
慢性过度使用常见药物，例如对乙酰氨基酚（扑热息痛），通常会导致急性肝衰竭（ALF）的发生。本研究旨在阐明培养间充质干细胞（MSCs）蛋白质组对乙酰氨基酚诱导的 ALF 动物肝损伤和再生动力学的影响，测试 MSCs 蛋白质组在细胞培养氧张力下的肝保护作用，并描绘负责该作用的蛋白质成分。从在轻度低氧（5%和 10%O 和正常（21%O）条件下培养的 MSCs 中制备的蛋白质组合物用于治疗通过注射对乙酰氨基酚诱导的小鼠 ALF。为了测试细胞培养中降低氧张力对所得 MSCs 蛋白质组含量的影响，我们应用高效液相色谱和质谱联用（LC-MS/MS）来鉴定在不同 O 水平下培养的 MSCs 中的蛋白质。用从培养的 MSCs 释放的蛋白质治疗接受对乙酰氨基酚治疗的动物，可抑制受损肝脏中的炎症反应；在最初 24 小时内减少肝细胞坏死的面积。从在较低 O 水平下培养的 MSCs 获得的组合物比从正常细胞培养的组合物更有效。通过细胞因子测定和蛋白质组学分析对蛋白质组合物进行的蛋白质图谱比较表征和个别成分的鉴定表明，即使是中度缺氧也会导致负责细胞内呼吸和细胞信号转导的各种蛋白质亚群的表达发生离散变化。在降低氧张力下体外培养的 MSCs 制备的蛋白质的应用可显著加速受损肝组织的愈合过程。从不同制剂获得的蛋白质组学数据为不同制剂提供了有关对培养基中氧张力敏感的 MSCs 蛋白质组中潜在候选物的新信息，这些候选物可能参与细胞用于应对急性肝衰竭的一般机制。
