Study On Hematopoietic Constituents In SI-WU-TANG By Chemomics Method

It is time and labor consuming to research, by "pure chemical separation-activity screening" method, the chemical foundation of efficacy of TCM drug, because its chemical composition is very complex. TCM Chemomics method is for the revelation, comparison and identification of multiple components in mixture from TCM drug. In the present study, Chemomics method was explored to study the hematopoietic constituents of SI-WU-TANG, a famous TCM hematopoietics, with the hope to find out a new method for the chemical foundation research of TCM drugs.Firstly SI-WU-TANG decoction was divided into three crude extracts A, B and C, and white blood cell (WBC) level in peripheral hemogram of radiated mice could be raised by extract B and extract C. Then B and C were divided further into Bl, B2, B3 and Cl, C2, C3. Activity analysis and Chemomics analysis were both carried out for all these 6 extracts. C2 and C3 were found to have better acivity of raising peripheral WBC. Chemomics analysis showed that the major constituents of C2 were monosaccharide, disaccharide and paeoniflorin, and the major constituents of C3 were monosaccharide and disaccharide. Monosaccharide and disaccharide contents in Bl, B2, and B3 were small. Cl had very little saccharide but various constituents of low or medium polarity such as paeoniflorin and ferulic acid. Therefore the hematopoietic activity was hypothetically related with paeoniflorin and mono/disaccharide. Then test of pure paeoniflorin showed that it could promote the proliferation of hematopoietic progenitor cell in bone marrow of radiated mice. On the other hand, C2 were fractionated into C2-1 (saccharide fraction) and C2-2 (non-saccharide fraction), and C2-1 showed better activity of both raising peripheral WBC and promoting the proliferation of hematopoietic progenitor cell. Further qualitative and quantitative analysis of C2 showed that monosaccharides in C2 were fructose and glucose, anddisaccharide should be sucrose. Then analytical grade fructose, glucose and sucrose were mixed together according to their contents in C2, and the saccharide-mixture showed activity of promoting the proliferation of hematopoietic progenitor cell. Therefore mono/disaccharide should be one of the active constituents in SI-WU-TANG. Furthermore, contents of fructose, glucose and sucrose in entire SI-WU-TANG decoction were determined and mixture of pure saccharides was prepared according to the contents. Then saccharide-mixture, SI-WU-TANG decoction, paeoniflorin, combinations of saccharide-mixture with C1, with B and with paeoniflorin were tested together. The result showed that all tested objects had activity of raising peripheral WBC and promoting the proliferation of hematopoietic progenitor cell. Activity of paeoniflorin itself had no significant difference with SI-WU-TANG decoction. The combination of saccharide-mixture with paeoniflorin had the best activity among the three combinations, which was better than that of SI-WU-TANG in individual index. All these demonstrated that paeoniflorin and mono/disaccharide were all hematopoietic constituents in SI-WU-TANG.