Near-infrared fluorescence imaging tool with large Stokes shift for sensitively detecting carboxylesterase 2 and monitoring its expression in non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) now affects more than one quarter of the global population and becomes a heavy public health burden. However, the underlying mechanism for the pathogenesis of NAFLD is still not clear. Carboxylesterase 2 (CES2), highly abundant in the liver and intestine, plays an important role in endogenous lipid metabolism and lipolysis. So far, the literatures for the role of CES2 in the development of NAFLD are still limited. In this study, we designed and synthesized a near-infrared fluorescent probe (HP-LZ-CES2) which can be specifically recognized and hydrolyzed by CES2, releasing a benzoate residue and a fluorophore (HP-LZ) with good fluorescence signal. With this probe, CES2 levels can be quantitatively measured in vitro and qualitatively visualized in living cells and mice. The probe has the advantages of large Stokes shift, high detection sensitivity and good selectivity. Further, the CES2 expression levels were visually investigated in both high-fat cells as the in vitro model for NAFLD and high-fat diet fed mouse as the in vivo model for NAFLD. The cell imaging experiments indicated a reduction of fluorescence signal in high-fat hepatic cells. The in vivo experiments showed an obvious reduction of fluorescence in the liver of NAFLD mouse model, which is consistent with the hepatic cell experiments. In contrast, an enhancement of fluorescence was observed in the intestine of NAFLD mouse model. As a result, the NAFLD mouse model can be visually distinguished from the normal chow mouse by vision. Therefore, the proposed probe can be an auxiliary tool for the diagnosis of NAFLD and a visual tool for understanding CES2's role in the development of NAFLD.