Evaluation of Various Anti-oketsu Kampo Formulae—Tokakujokito,Keishibukuryogan and Tokishakuyakusan—for Microcirculation by Live Imaging / 日本東洋医学雑誌

Oketsu is a characteristic pathophysiology in Kampo and traditional East Asian medicine that includes mul­tiple aspects of hemodynamic disorder. Anti­-oketsu drugs or the Kampo formulation used for oketsu show sig­nificant clinical effects on various disorders; however, their underlying mechanisms still remain unclear. We aimed to clarify the characteristics of the pharmacological effects of anti-­oketsu drugs on the microcirculation using a microscopic live imaging technique. Three Kampo formulations, namely tokakujokito, keishibukuryo­gan, and tokishakuyakusan were orally administrated to C57BL/6 mice at a dose of 300 mg/kg diluted in dis­tilled water. Live imaging was performed on the subcutaneous vessels of the mice, including the arteries (di­ameter > 50 μm), arterioles (diameter 10-50 μm) and capillaries (diameter < 10 μm). Tokakujokito widely increased erythrocyte flow velocity and blood flow volume from arteries to capillaries within 60 min of ad­ministration. The effects of keishibukuryogan on the vasodilation of the arterioles were remarkable, and con­tinued up to 120 min after administration. The pharmacological target of tokishakuyakusan was the capillaries, increasing their erythrocyte velocity and blood flow volume;its effect was more slowly expressed than those of the other formulations. Our results clearly demonstrate the sequential and special effects of anti-­oketsu drugs on hemodynamics. These differences may provide pharmacological information on the clinical usage of traditional Kampo formulations.

Molecular mechanisms of DHEA and DHEAs on apoptosis and cell cycle arrest via Akt pathway in hepatoma cell lines / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the anti-proliferation effects of dehydroepiandrosterone (DHEAéand DHEA sulfate (DHEAs) on tumor cells.</p><p><b>METHODS</b>Human hepatoblastoma cells (HepG2) and colon adenocarcinoma cells (HT-29) were treated with DHEA and DHEAs of various concentrations. The cells were incubated for 8, 24, 48, and 72 hours, and the proliferation, apoptosis, cell cycle and the expression of phosphorylated Akt (Thr308 and Ser473) were analyzed using MTT assay, flow cytometry, and Western blotting at different time points. The influences of an inhibitor (LY294002) and an activator (hepatic growth factor; HGF) of PI3K on the effectiveness of DHEA were determined in HepG2 cells.</p><p><b>RESULTS</b>By increasing the concentrations of DHEA (1, 10, 50, 100, 200 micromol/L), the percentages of HepG2 and HT-29 survival cells treated with DHEA at 24 h were 92.7%+/-0.9%, 84.7%+/-1.2%, 62.4%+/-0.8%, 49.5%+/-0.8%, 50.7%+/-0.3% and 92.5%+/-0.4%, 89.5%+/-0.7%, 80.5%+/-1.1%, 67.5%+/-1.5%, 70.6%+/-0.6%, respectively. Proliferations of HepG2 and HT-29 cells were significantly inhibited after 24 hours of being incubated with 100 micromol/L DHEA treatment; the inhibition effect was stronger on HepG2 cells than on HT-29 cells. The effect of DHEAs on both cell lines on cell proliferation was weaker than that of the DHEA. In the cell cycle assay, DHEA treatment induced cell arrest in G0/G1 phase in both cell lines. Apoptosis of HepG2 cells was significantly triggered (18.6%+/-2.2%) by 100 micromol/L DHEA treatment for 24 hours, but not by DHEAs. In addition, 100 and 200 micromol/L DHEA treatments for 24 hours markedly inhibited phosphorylations of Akt (Thr308 and Ser473) in HepG2 cells, and these effects were enhanced by exposing them to LY294002 and stopped by exposing them to HGF. The anti-proliferative effects of DHEA on tumor cell lines were much stronger than those of DHEAs, and they were even stronger in HepG2 cells than in HT-29 cells.</p><p><b>CONCLUSION</b>Our results suggest that the induction of apoptosis through the inhibition of Akt signaling pathway is one of the anti-proliferative mechanisms of DHEA in certain tumors, but DHEA also promotes cell-cycle arrest.</p>