Profiling of 120 types of herbal extracts and their effects on morphology in cultured neuronal or glial cell lines, followed by RNA extraction for a cDNA library: Consideration for use in studies based on Kampo theories

Abstract

Aim Kampo medicine is composed of a variety of crude drugs. Each crude drug has its own effects, and the interaction of all drugs produces the therapeutic effect of Kampo medicine. In order to examine the characteristics of Kampo medicine and its crude drugs, 120 types of herbal medicines were characterized based on their three‐dimensional high‐performance liquid chromatography (3D‐HPLC) profiles as well as their effects on cell morphology/proliferation when added to cultured cells and gene expression changes such as on BNIP‐3 mRNA levels as an example. Methods The cultured neural cell line N18‐TG‐2 and glial cell line C6Bu‐1 were used in experiments. After the acquisition of 3D‐HPLC data, each crude drug was added to the cell culture medium at concentrations of 10 and 100 μg/mL, images were taken of morphological changes, and RNA was extracted to prepare a complementary DNA (cDNA) library. Results All the results obtained from the treatments with 120 crude drugs are listed in the Supporting Information. Based on these data, as an example the expression of antidepressant‐related factor BNIP‐3, which we had previously found to increase in expression after treatment with Hochuekkito, was studied and the results obtained revealed that it was increased by Astragali Radix, Cimicifugae Rhizoma, Glycyrrhizae Radix and others. Conclusion The present results, which were collected from observations under unified conditions, include information that will promote herbal medicine research. Future studies to examine the effects of crude drug combinations are also discussed, suggesting that the information files and cDNA library presented here can be the basis for many future integrated studies such as new Wakan‐yaku prescription developments.

Effects of herbal extracts on mRNA expression of intrinsic antidepressant‐related candidate factor BNIP‐3 in cultured neuronal and glial cell lines: Relationship with the Kampo concept of Qi

“… Note : The data in this table were extracted from our previously reported findings [11]. The top shows the results from C6 and the bottom from N18.…”

“…We have previously reported that RNA was extracted from a cultured neuronal cell line N18TG‐1 (N18) and glial cell line C6Bu‐1 (C6) treated with 120 kinds of herbal extracts which were supplied by the Institute of Natural Medicines (INM), University of Toyama, Japan. Using these RNAs, a cDNA library was prepared [11]. Furthermore, using the cDNA library, we also examined all of the 120 crude drug extracts from the overall perspective to determine which herbal drugs are involved in BNIP‐3 mRNA changes [11].…”

“…The top 30 crude drugs in terms of the BNIP-3 expression in order of increasing or decreasing BNIP-3 mRNA expression. The data in this table were extracted from our previously reported findings [11]. The top shows the results from C6 and the bottom from N18.…”

“…Using these RNAs, a cDNA library was prepared [11]. Furthermore, using the cDNA library, we also examined all of the 120 crude drug extracts from the overall perspective to determine which herbal drugs are involved in BNIP-3 mRNA changes [11].…”

Examination of the molecular mechanism of the mutual inhibitory effect of each cytotoxicity caused by the combined extraction of Coptidis Rhizoma and Rhei Rhizoma: An example of providing evidence for ‘herbal pair’ theory

“…The authors created a cDNA library for the purpose of examining the gene expression effect of 120 kinds of crude drugs alone [2], and are proceeding with various experiments on the components' cytotoxicity, function, and gene expression. In the process, we observed a mysterious phenomenon related to CR and RR.…”