Wang Xiao-Xiao, Jin Rui, Li Xiao-He, Yang Qiang, Teng Xiao, Liu Fang-Fang, Wu Nan, Rao Hui-Ying, Liu Feng
Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing International Cooperation Base for Science and Technology on NAFLD Diagnosis, Beijing, China.
Hangzhou Choutu Technology Co., Ltd., Hangzhou, China.
Front Med (Lausanne). 2023 Jun 2;10:1172058. doi: 10.3389/fmed.2023.1172058. eCollection 2023.
Non-alcoholic fatty liver disease (NAFLD) is a global commonly occurring liver disease. However, its exact pathogenesis is not fully understood. The purpose of this study was to quantitatively evaluate the progression of steatosis and fibrosis by examining their distribution, morphology, and co-localization in NAFLD animal models.
Six mouse NAFLD groups were established: (1) western diet (WD) group; (2) WD with fructose in drinking water (WDF) group; (3) WDF + carbon tetrachloride (CCl4) group, WDF plus intraperitoneal injection of CCl4; (4) high-fat diet (HFD) group, (5) HFD with fructose (HFDF) group; and (6) HFDF + CCl4 group, HFDF plus intraperitoneal injection of CCl4. Liver tissue specimens from NAFLD model mice were collected at different time points. All the tissues were serially sectioned for histological staining and second-harmonic generation (SHG)/two-photon excitation fluorescence imaging (TPEF) imaging. The progression of steatosis and fibrosis was analyzed using SHG/TPEF quantitative parameters with respect to the non-alcoholic steatohepatitis Clinical Research Network scoring system.
qSteatosis showed a good correlation with steatosis grade (: 0.823-0.953, < 0.05) and demonstrated high performance (area under the curve [AUC]: 0.617-1) in six mouse models. Based on their high correlation with histological scoring, qFibrosis containing four shared parameters (#LongStrPS, #ThinStrPS, #ThinStrPSAgg, and #LongStrPSDis) were selected to create a linear model that could accurately identify differences among fibrosis stages (AUC: 0.725-1). qFibrosis co-localized with macrosteatosis generally correlated better with histological scoring and had a higher AUC in six animal models (AUC: 0.846-1).
Quantitative assessment using SHG/TPEF technology can be used to monitor different types of steatosis and fibrosis progression in NAFLD models. The collagen co-localized with macrosteatosis could better differentiate fibrosis progression and might aid in developing a more reliable and translatable fibrosis evaluation tool for animal models of NAFLD.
非酒精性脂肪性肝病（NAFLD）是一种全球常见的肝脏疾病。然而，其确切发病机制尚未完全明确。本研究的目的是通过检查NAFLD动物模型中脂肪变性和纤维化的分布、形态及共定位情况，对其进展进行定量评估。
建立六个小鼠NAFLD组：（1）西式饮食（WD）组；（2）饮用水中添加果糖的WD（WDF）组；（3）WDF + 四氯化碳（CCl4）组，即WDF组加腹腔注射CCl4；（4）高脂饮食（HFD）组；（5）含果糖的HFD（HFDF）组；（6）HFDF + CCl4组，即HFDF组加腹腔注射CCl4。在不同时间点收集NAFLD模型小鼠的肝组织标本。所有组织均进行连续切片用于组织学染色和二次谐波产生（SHG）/双光子激发荧光成像（TPEF）成像。根据非酒精性脂肪性肝炎临床研究网络评分系统，使用SHG/TPEF定量参数分析脂肪变性和纤维化的进展情况。
qSteatosis与脂肪变性分级显示出良好的相关性（：0.823 - 0.953，< 0.05），并且在六个小鼠模型中表现出高性能（曲线下面积[AUC]：0.617 - 1）。基于与组织学评分的高度相关性，选择包含四个共享参数（#LongStrPS、#ThinStrPS、#ThinStrPSAgg和#LongStrPSDis）的qFibrosis来创建一个能够准确识别纤维化阶段差异的线性模型（AUC：0.725 - 1）。与大泡性脂肪变性共定位的qFibrosis通常与组织学评分的相关性更好，并且在六个动物模型中的AUC更高（AUC：0.846 - 1）。
使用SHG/TPEF技术进行定量评估可用于监测NAFLD模型中不同类型的脂肪变性和纤维化进展。与大泡性脂肪变性共定位的胶原蛋白能够更好地区分纤维化进展，可能有助于为NAFLD动物模型开发更可靠且可转化的纤维化评估工具。
