| Objective Sanguisorba officinalis is a traditional chinese medicine which can cool blood hemostasis and detoxification boils.At the same time,its white cells-lifting effect has been sured with the wide application of Diyushengbaipian.And it can be effective for a small dose,equate with 60mg medicinal materials.But Diyushengbaipian is a single flavor powder,and we found many problems in the course of clilic application,such as contradictions of does-response,material basis and mechanism were unknown.Besides,as a traditional medicine,it caused the slow and poor effect.To solve those problems,our project expand a systematic study in lifting the white cells.At the same time,it can provide a theoretical basis for a new drug can rise the white blood cells enduringly and qucikly and protecte the bone marrow.Mehods First,we destermine the effective parts by gavaging mice which has been leukopenia caused by cyclophamide the treated sanguisorba extract;Then we deal the effective parts by the different concentration of ethanol,and screen effective components for animal reseach by determinated the WBS;Next,we separated and preparated the compounds from effective components by polyamide, silicagel, sephadex LH-20 and ODS C18 colum chromatography,and dotermine the compounds structure through physical,chemical constants and spectrum analysis technology combined with literature;Finally,we confirm the effectives of the compounds by the M-NFS-60 cell lines with the methods of MTT and CCK-8.Results Through screening step by step.we have found the effective parts increased the white cells that included ethyl aeetate and water area.and after eluted the effectivion parts,we sure that the 60% and 95% methanol eluate from ethyl acetate parts are effective components;and nine compounds were preparated,separated and identified from the 60%methanol eluate which has the best activity.The nine components are all triterpene composition,including 3?,19?-dihydroxyurs-12-en-28-oic acid 28-?-D-glucopyranoside(component I); 2?,3?,19a-trihydroxyurs-12-en-28- oic acid28-?-D-galactopyranosyl ester(eomponent ?); 3?-[(a-L-Arabinopyranosyl)oxy]-urs-12,18-dien-28-oic acid 28-?-D-g lucopyranosyl ester(component ?); 3?,19?-hydroxyolean-12-en-28-oic acid 28-?-D-glucopyrano-side(component ?); kaji-ichigosideFl(component V); ziyuglycoside-?(component VI); 3?-[(?-L-Arabinopyraaosyl)oxy]-23-hydr oxyurs-12,19(29)-dien-28-oic acid28-?-D-glucopyranosyl ester(component ?); 3?-[(a-L-Arabinopyranosyl)oxy]-urs-11, 13(18)-dien-28-oic acid 28-P-D-glueopyranosyl ester(component VIII); ziyu glycoside ?(component IX).Through the cell experiment,we found that the compounds of VI and ? can rise the white blood cells significantly.Conclusion In our project,the nine compounds preparated combined system solvent method with the active tracking are all saponins.It shows that saponins can increase the numbers of peripheral white blood cells.Meanwhile,we have define the material basis in rising the white blood cells of sanguisorba.By analysis the structure,we concluded that the active groups include carboxyl groups of 17,glyeoside of 3,hydroxyl groups of 19;The saponins of Sanguisorba is mainly applied to the middle part in the bone marrow,and rise the white blood cells by promoting expression of MGMT and protein,promoting the proliferation and differentiation of granulocyte.Based on the previous research discovers,we laid the foundation and provided a research basis on sompatibility Sanguisorba tannins and saponins. |
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