It has been reported that the expression level of BNIP-3 mRNA increases by antidepressants and Hochu-ekki-To (HET), suggesting that BNIP-3 may be a candidate depression-related factor. Therefore, in order to examine the relationship between BNIP-3 and Qi, the expression levels of BNIP-3 mRNA were examined using 22 newly extracted crude drugs which were selected from the previous library results on 120 crude drugs in the neuronal cell line N18TG-2 and in the glial cell line C6Bu-1. Methods: Based on the results on 120 kinds of crude drugs in a cDNA library, 22 kinds were selected and newly extracted, and then cells were re-treated with them for a secondary assay by real-time polymerase chain reaction (PCR) methods. Results: Many of the herbal medicines contained in HET increased BNIP-3 mRNA expression overall. On the other hand, in the results with especially the neuronal cell line N18TG-2, the herbal medicines that make up Oren-gedoku-To (OGT) and San-o-shashin-To (SOST) strongly showed suppressive effects on the expression. Conclusion: Since HET clinically has the effect of raising mood, while an effect of OGT and SOST is to suppress the rise of mood, the result of this study may be of interest regarding the link between control of the direction of Qi and the expression of BNIP-3 mRNA.

Aim: Kampo medicines are effective depending on the combination of crude drugs. In addition, the basis is a combination of two crude drugs, which is defined as a 'herbal pair'. However, there are very few studies from this point of view. We found that Rhei Rhizoma (RR) may interact with Coptidis Rhizoma (CR). Methods: Component profiles were analyzed by three-dimensional high-performance liquid chromatography (3D-HPLC). Cell viability in N18TG-2 cells was determined by morphology observation and a '3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, yellow tetrazole (MTT)' assay. Results: When RR and CR were extracted together, the color of the extract was lighter than the colors of the individual extracts. In addition, precipitation occurred by simply combining the extracts of both in vitro. When each component profile was analyzed by 3D-HPLC, at least the amount of berberine, which is contained in a large amount in CR, was strongly decreased in combined extracts with RR. Although CR and RR each have a cytotoxic effect, when both were extracted together, their cytotoxicity almost disappeared. However, cytotoxicity remained in the supernatant when both extracts were combined in vitro, and also in the suspension of the resulting precipitate. Conclusion: From these results, when CR and RR are extracted together, their cytotoxicity is suppressed by some unknown mechanism. Whether this is due to the synthesis of new compounds by co-extraction or the detoxification of toxic components by interaction needs further investigation. This result suggests the importance of 'herbal pair' research.