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. 2023 Feb 2;12(3):479. doi: 10.3390/cells12030479.
To reduce the risk of post-hepatectomy liver failure in patients with hepatic pathologies, it is necessary to develop an approach to express the intraoperative assessment of the liver's regenerative potential. Traditional clinical methods do not enable the prediction of the function of the liver remnant. Modern label-free bioimaging, using multiphoton microscopy in combination with second harmonic generation (SHG) and fluorescence lifetime imaging microscopy (FLIM), can both expand the possibilities for diagnosing liver pathologies and for assessing the regenerative potential of the liver. Using multiphoton and SHG microscopy, we assessed the structural state of liver tissue at different stages of induced steatosis and fibrosis before and after 70% partial hepatectomy in rats. Using FLIM, we also performed a detailed analysis of the metabolic state of the hepatocytes. We were able to determine criteria that can reveal a lack of regenerative potential in violated liver, such as the presence of zones with reduced NAD(P)H autofluorescence signals. Furthermore, for a liver with pathology, there was an absence of the jump in the fluorescence lifetime contributions of the bound form of NADH and NADPH the 3rd day after hepatectomy that is characteristic of normal liver regeneration. Such results are associated with decreased intensity of oxidative phosphorylation and of biosynthetic processes in pathological liver, which is the reason for the impaired liver recovery. This modern approach offers an effective tool that can be successfully translated into the clinic for express, intraoperative assessment of the regenerative potential of the pathological liver of a patient.
为了降低肝疾病患者肝切除术后肝功能衰竭的风险，有必要开发一种方法来表达术中对肝脏再生潜能的评估。传统的临床方法无法预测肝剩余功能。现代无标记的生物成像技术，结合多光子显微镜、二次谐波产生（SHG）和荧光寿命成像显微镜（FLIM），既可以扩展诊断肝疾病的可能性，也可以评估肝的再生潜能。我们使用多光子和 SHG 显微镜，在大鼠 70%部分肝切除术前和术后评估了诱导性脂肪变性和纤维化的不同阶段肝组织的结构状态。我们还使用 FLIM 对肝细胞的代谢状态进行了详细分析。我们能够确定可以揭示受损肝脏再生潜能不足的标准，例如 NAD(P)H 自发荧光信号减少的区域。此外，对于有病理学的肝脏，在肝切除术后第 3 天，结合态 NADH 和 NADPH 的荧光寿命贡献没有出现跃升，这是正常肝再生的特征。这些结果与病理肝脏中氧化磷酸化和生物合成过程的强度降低有关，这是肝恢复受损的原因。这种现代方法提供了一种有效的工具，可以成功地转化为临床，用于快速评估患者病理性肝脏的再生潜能。
