Exploring the microscopic changes of lipid droplets and mitochondria in alcoholic liver disease via fluorescent probes with high polarity specificity

Alcoholic liver disease (ALD) has received extensive attention because of the increasing alcohol consumption globally as well as its high morbidity. It is reported that absorbed alcohol can cause lipid metabolism disorder and mitochondria dysfunction, so here in this work, we planned to study the microscopic changes of the two organelles, lipid droplets (LDs) and mitochondria in hepatocyte, under the stimulation of alcohol, hoping to present some meaningful information for the theranostics of ALD by the technique of fluorescence imaging. Guided by theoretical calculation, two fluorescent probes, named CBu and CBuT, were rationally designed. Although constructed by the same chromophore scaffold, they stained different organelles efficiently and emitted distinctively. CBu with high lipophilicity, ascribed to the two butyl groups, can selectively localize in LDs with green fluorescence, while CBuT bearing a triphenylphosphine unit can specifically target mitochondria due to electrostatic interactions with near-infrared (NIR) fluorescence. Both probes displayed remarkable selectivity and sensitivity to polarity, free from the environmental interferences including viscosity, pH and other bio-species. With these two probes, the accumulation of LDs and polarity decrease in mitochondria were clearly monitored at the green and red channels, respectively, in the ALD cell model. CBuT was further applied to image the mice with ALD in vivo. In short, we have confirmed the valuable organelles, LDs and mitochondria, for ALD study and provided two potent molecular tools to visualize their changes through fluorescence imaging, which would be favorable for the further development of theranostics for ALD.