Sakaguchi S, Furusawa S, Yokota K, Sasaki K, Takayanagi Y
First Department of Hygienic Chemistry, Tohoku College of Pharmacy, Sendai, Japan.
Biol Pharm Bull. 1995 Apr;18(4):621-3. doi: 10.1248/bpb.18.621.
The present study investigated whether or not Sho-saiko-to (crude powder extract, TJ-9) can suppress nitric oxide (NO) generation by endotoxin-activated J774A.1 cells in order to study the preventive mechanism of Sho-saiko-to against endotoxemia. In this experiment, we estimated the NO2- in the murine macrophage cell line J774A.1 using the Griess method. Our results clearly demonstrated that J774A.1 cells stimulated with endotoxin (0.01-10 micrograms/ml) can effectively produce NO, and the production was dependent on the dose of endotoxin. On the other hand, we investigated the suppressive effect of TJ-9 (10-100 micrograms/ml) on NO generation by endotoxin (0.1 microgram/ml)-activated J774A.1 cells. The NO level when the cells were incubated with endotoxin and TJ-9 (10-20 micrograms/ml) was slightly lower than that in cells treated with endotoxin alone. In contrast, treatment with TJ-9 (50-100 micrograms/ml) significantly inhibited endotoxin-activated NO generation in J774A.1 cells, whereas the treatment with TJ-9 (10-100 micrograms/ml) alone was ineffective in inducing NO formation and in inhibiting cell viability in the J774A.1 cells. These findings suggest that a Kampo presciption of Sho-saiko-to shows a suppressive effect on NO generation in macrophages stimulated with endotoxin, and that it may be useful in improving endotoxin-shock symptoms.
本研究旨在探讨小柴胡汤（粗粉提取物，TJ - 9）是否能够抑制内毒素激活的J774A.1细胞产生一氧化氮（NO），以此研究小柴胡汤预防内毒素血症的机制。在本实验中，我们采用格里斯方法测定小鼠巨噬细胞系J774A.1中的NO₂⁻。我们的结果清楚地表明，用内毒素（0.01 - 10微克/毫升）刺激的J774A.1细胞能够有效地产生NO，且其产生量依赖于内毒素的剂量。另一方面，我们研究了TJ - 9（10 - 100微克/毫升）对经内毒素（0.1微克/毫升）激活的J774A.1细胞产生NO的抑制作用。当细胞与内毒素和TJ - 9（10 - 20微克/毫升）一起孵育时，NO水平略低于仅用内毒素处理的细胞。相反，用TJ - 9（50 - 100微克/毫升）处理可显著抑制J774A.1细胞中内毒素激活的NO生成，而单独用TJ - 9（10 - 100微克/毫升）处理在诱导J774A.1细胞中NO形成及抑制细胞活力方面无效。这些发现表明，汉方制剂小柴胡汤对内毒素刺激的巨噬细胞中NO的生成具有抑制作用，并且可能有助于改善内毒素休克症状。
