Generation of reactive oxygen species and induction of apoptosis of HL60 cells by ingredients of traditional herbal medicine, Sho-saiko-to.

The prooxidant and apoptosis-inducing effects of Sho-saiko-to, a traditional Sino-Japanese herbal medicine and its active ingredients were analyzed. Among the components of Sho-saiko-to, wogon, the extract of Scutellaria and licorice root extract induced apoptosis of HL60 cells and increased the intracellular levels of reactive oxygen species. Lower concentrations (5 to 20 muM) of baicalein, the principal flavonoid in the Scutellaria root extract, showed induction of cell apoptosis and elevated the intracellular reactive oxygen species. However, the increase in the concentrations of baicalein rather inhibited the induction of apoptosis and the elevated levels of reactive oxygen species in cells. Induction of baicalein-mediated apoptosis was inhibited by addition of Tempol, the scavenger of reactive oxygen species. Glycyrrhetinic acid, an ingredient of licorice root extract, also induced apoptosis followed by increase in the intracellular reactive oxygen species. The effect of Sho-saiko-to on cell differentiation can be explained by the action of two ingredients, baicalein and glycyrrhetinic acid, which cause apoptosis and increase in reactive oxygen species in cells.

Regulatory role of polyamine in the acid phosphatase from potato tubers.

Effects of polyamine and metal ions on the new type of acid phosphatase purified from potato (Solanum tuberosum L. Irish Cobbler) tubers were analyzed. The enzyme belongs to nonspecific acid phosphatase family (EC 3.1.3.2), which hydrolyzes various phosphorylated substrates. The enzyme hydrolyzed inorganic pyrophosphate as a preferred substrate, and exhibited the hyperbolic kinetics with respect to the substrate, inorganic pyrophosphate in the absence of metal cations. Polyamine activated the enzyme effectively by lowering the K(m) value without appreciable changes in the maximal velocity. The most effective polyamines as activators were spermine and spermidine. Mg(2+) ion increased the K(m) value without affecting the maximal velocity of the enzyme, but Ca(2+) ion decreased both the K(m) and V(max) values. Increasing concentrations of spermine also decreased the K(m) value irrespective of Mg(2+) ion included, but gave a constant K(m) and V(max) values in the absence and presence of Ca(2+) ion. Action of spermine and metal ions can be explained by the complex formation with the substrate pyrophosphate. The acid phosphatase from potato can utilize the pyrophosphate-spermine or pyrophosphate-Ca(2+) complex as the preferred substrates. However, the enzyme can use the pyrophosphate-Mg complex with a weak affinity for the active site. Polyamine activates acid phosphatase in the absence and presence of metal cations, and activation by polyamine of the enzyme may contribute to the stimulation of starch biosynthesis and the control of glycolysis/gluconeogenesis by regulating PPi levels in growing potato tubers.