Role of Reactive Oxygen Species on Sodium Butyrate Induced Human Hepatocyte Differentiation

Reactive oxygen species (ROS) such as hydrogen peroxide, superoxide anion and hydroky radicals are produced in various physiologic and pathologic conditions and involved in many cellular processes as proliferation, differentiation and apoptosis. Studies investigating the role of ROS in various cellular behaviors especially in proliferation and apoptosis have been widely conducted in many cell types but the role of ROS in nontransformed human hepatocyte differentiation has not been investigated yet. thus we were going to elucidate the roleof ROS on human hepatocyte differentiaiton using sodium butyrate (SB) induced hepatocyte differentiation model of our own establishment.

Intracellular ROS and apoptotic cell death were monitored by flowcytometry using peroxide sensitive probe (Dicholorofluorscein diacetate) and Annexin V/Propidium iodide, respectively. Urea nitrogen in culture media was measured by colorimetric methods. Ornithine transcarbamylase(OTC) and albumin trasncription was evaluated by RT-PCR.

Intracellular ROS production was increased by SB. SB induced urea production was significantly decreased with antioxidant treatment (p<0.05) and SB induced OTC and albumin transcription were also attenuated with antioxidant treatment. SB induced increase in apoptosiswas significantly inhibited by antioxidant treatment (p<0.05).

ROS produced during the process of sodium butyrate induced human hepatocyte differentiation auguments hepatoctye differentiation and apoptosis.