กลไลปกป้องของฟิแลนด์ทินต่อความเป็นพิษของแอลกอฮอล์ที่ต่อเซลล์เพาะเลี้ยงเฮพ จี 2 และเซลล์เพาะเลี้ยงร่วมเฮพจี 2 และ ทีเฮซ พี 1

Abstract

Alcohol has been considered to be a major risk factor related to liver injury. Phyllanthin is a hepatoprotective agent that has been reported to reduce hepatotoxicity initiated by various compounds. Thus, our study aimed to evaluate the effects and possible mechanisms of phyllanthin on alcohol induced liver injury in HepG2 cells as well as with a co-culture of HepG2 cells with THP-1 differentiated macrophage cells. Treatment of cells with alcohol for 4 and 72 h resulted in a significant decrease inviability for both cell types, and we further proved that such toxicity was triggered by production of reactive oxygen species (ROS), changes to mitochondrial membrane potential (MMP) and apoptotic cell death. Alcohol (4 and 72 h) exposure significantly increased the intracellular ROS generation, decreased the MMP changes, increased the number of apoptotic and necrotic cells and also induced caspase-3/7 activity in a co-culture with THP-1 differentiated macrophage cells higher than HepG2 cells alone. Pretreatment of the HepG2 cells with phyllanthin for 24 hour prior to alcohol exposure for 4 h significantly decreased the intracellular production of ROS and also improved the change of the MMP as well as causing a decrease of the number of apoptotic cells and inhibited the caspase-3/7 activity. In the co-cultured cells, phyllanthin was also shown to attenuate the increase of ROS production, increased the MMP at the highest concentration of phyllanthin and also improved cell survival by inhibiting the caspase-3/7 activity. In contrast, pretreatment of the HepG2 cells with 10 μM phyllanthin for 24 hour prior to alcohol exposure for 72 h significantly decreased the intracellular production of ROS and also improved the change of the MMP as well as causing a decrease in the number of apoptotic cells and inhibited the caspase-3/7 activity but not affect to the co-cultured cells. Our results clearly indicated that treatment with phyllanthin should have a significant therapeutic effect on alcohol-related liver diseases.