Institute of Experimental Oncology and Biomedical Technologies, Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Sq., 603000 Nizhny Novgorod, Russia.
Department of Biophysics, N.I. Lobachevsky Nizhny Novgorod National Research State University, 23 Gagarina Ave., 603022 Nizhny Novgorod, Russia.
Cells. 2021 Oct 26;10(11):2894. doi: 10.3390/cells10112894.
The search for new criteria indicating acute or chronic pathological processes resulting from exposure to toxic agents, testing of drugs for potential hepatotoxicity, and fundamental study of the mechanisms of hepatotoxicity at a molecular level still represents a challenging issue that requires the selection of adequate research models and tools. Microfluidic chips (MFCs) offer a promising in vitro model for express analysis and are easy to implement. However, to obtain comprehensive information, more complex models are needed. A fundamentally new label-free approach for studying liver pathology is fluorescence-lifetime imaging microscopy (FLIM). We obtained FLIM data on both the free and bound forms of NAD(P)H, which is associated with different metabolic pathways. In clinical cases, liver pathology resulting from overdoses is most often as a result of acetaminophen (APAP) or alcohol (ethanol). Therefore, we have studied and compared the metabolic state of hepatocytes in various experimental models of APAP and ethanol hepatotoxicity. We have determined the potential diagnostic criteria including the pathologically altered metabolism of the hepatocytes in the early stages of toxic damage, including pronounced changes in the contribution from the bound form of NAD(P)H. In contrast to the MFCs, the changes in the metabolic state of hepatocytes in the ex vivo models are, to a greater extent, associated with compensatory processes. Thus, MFCs in combination with FLIM can be applied as an effective tool set for the express modeling and diagnosis of hepatotoxicity in clinics.
寻找新的标准来指示由于暴露于有毒物质而导致的急性或慢性病理过程，测试药物的潜在肝毒性，以及在分子水平上研究肝毒性的机制，这些仍然是一个具有挑战性的问题，需要选择合适的研究模型和工具。微流控芯片 (MFC) 为快速分析提供了有前途的体外模型，并且易于实现。然而，为了获得全面的信息，需要更复杂的模型。一种研究肝病理的全新无标记方法是荧光寿命成像显微镜 (FLIM)。我们获得了与不同代谢途径相关的游离和结合形式的 NAD(P)H 的 FLIM 数据。在临床病例中，过量引起的肝病理最常见的是由于对乙酰氨基酚 (APAP) 或酒精 (乙醇)。因此，我们研究并比较了各种 APAP 和乙醇肝毒性实验模型中肝细胞的代谢状态。我们确定了潜在的诊断标准，包括在有毒损伤的早期阶段肝细胞病理性改变的代谢，包括结合形式的 NAD(P)H 的贡献明显变化。与 MFC 相比，离体模型中肝细胞代谢状态的变化在更大程度上与代偿过程有关。因此，MFC 与 FLIM 结合可以作为临床中肝毒性快速建模和诊断的有效工具集。
