EFFECTS OF HYDROGEN PEROXIDE ON PRODUCING INTRACELLULAR ROS OF WB-344 RAT LIVER EPITHELIAL CELL
H2O2致WB-F344细胞内活性氧的产生及机理

To study the role and mechanism of hydrogen peroxide(H 2 O 2 ) on intracellular reactive oxygen species(ROS) generation of WB-F344 rat liver epithelial cell, DHR123 was used as fluorescence probe to detect the intracellular ROS generation and observed on laser scanning confocal microscope. The results showed that: (1)The generation of intracellular ROS of the cell stimulated by H 2 O 2 increased slowly and gradually. (2)The intracellular ROS production stimulated by H 2 O 2 decreased when pretreated with ROS scavenger NAC. (3)There were no production of intracellular ROS pretreated with 2-Aminop-urine, a broader spectrum protein kinase inhibitor, or pretreated with Tyrphostin 25, a specific inhibitor of tyrosine protein kinase, or with Bisindolylmaleimide I, a PKC inhibitor. (4)The intracellular ROS could still be generated but the level decreased slightly by incubated in D-Hanks solution containing EG-TA, the chelate of extracellular Ca 2+ . (5)There were no generation of intracellular ROS pretreated with NAC and incubated WB cell in D-Hanks solution containing EGTA. These results suggested that the pathway of intracellular ROS generation stimulated by H 2 O 2 may be related to PKC, TPK and Ca 2+ signal transduction.