MicroRNA-200c coordinates HNF1 homeobox B and apolipoprotein O functions to modulate lipid homeostasis in alcoholic fatty liver disease

miR-141/200c contributes to ethanol-mediated hepatic glycogen metabolism

“…In our previous study where we used the same ethanol feeding regimen, we found that miR-141 and miR-200c levels were significantly induced by chronic ethanol feeding [ 35 ]. The seed sequences of miR-141 and miR-200c in mice are identical to those in humans.…”

“…We used male mice in all the experiments. Eight- to ten-week-old mice were subjected to the ethanol feeding regimen described previously ( Supplementary Figure 1 ) [ 35 , 36 ]. Briefly, mice were fed the control Lieber–DeCarli liquid diet (Bio-Serv F1259SP) for 5 days to acclimatize to liquid tube feeding, followed by feeding the same diet continuously for a control group (Ctrl) or the Lieber–DeCarli liquid diet (Bio-Serv F1258SP) supplemented with ethanol (5% vol/vol) for an ethanol group (EtOH).…”

“…Hepatocytes were isolated from eight- to ten-week-old mice by two-step collagenase perfusion, as described previously [ 35 ], with minor modifications. In brief, mice were fasted overnight and anaesthetized by intraperitoneal (IP) injection of ketamine and xylazine.…”

“…Liquid chromatography coupled to mass spectrometry (LC/MS)-based metabolomics studies were performed for untargeted screening of hepatic metabolome. 50 mg of snap frozen liver samples from WT and KO were processed at the West Coast Metabolomics Center, University of California, Davis, as described previously [ 35 ]. Bioinformatic data analysis was performed using MetaboAnalyst 3.0/5.0.…”

“…Studies have shown that miR-200 members are involved in cancer development and progression [ 29 ], female fertility [ 30 ], tooth development [ 31 ], and type 2-diabetes associated pancreatic beta cell survival [ 32 ]. Recently, our studies have demonstrated that the miR-141 and -200c cluster (hereafter referred to as miR-141/200c) plays critical roles in nonalcoholic steatohepatitis [ 33 ], cholestatic liver injury [ 34 ], and alcoholic fatty liver disease [ 35 ]. However, whether miR-141/200c influences glucose homeostasis during chronic ethanol feeding remains largely elusive.…”