Development of an in vitro cell culture model of hepatic steatosis using hepatocyte-derived reporter cells

摘要: Fatty liver disease is a problem of growing clinical importance due to its association with the increasingly prevalent conditions obesity and diabetes. While steatosis represents reversible state excess intrahepatic lipid, it also associated increased susceptibility oxidative cytokine stresses progression irreversible hepatic injury characterized by steatohepatitis, cirrhosis, malignancy. Currently, molecular mechanisms underlying this dynamic remain poorly understood, particularly at level transcriptional regulation. We recently constructed library stable monoclonal green fluorescent protein (GFP) reporter cells that enable regulation be studied dynamically in living cells. Here, we adapt create model will allow investigation dynamics development response subsequent “second hit” stresses. The recapitulates many cellular features human disease, including fatty acid uptake, intracellular triglyceride accumulation, reactive oxygen species decreased mitochondrial membrane potential, apoptotic stresses, proliferation. Finally, demonstrate utility for studying regulation, compared nuclear factor κB (NFκB), heat shock element (HSE), glucocorticoid (GRE) their classical inducers under lean found lipid accumulation was dose-dependent impairment NFκB HSE but not GRE activation. Thus, steatotic represent an efficient responses have potential provide important insights into disease.