Studies On The Quality Evaluation Of Four Ginseng Pills And The Antitumor Effect Of Renshen Guipi Granule

Ginseng(Panax ginseng C.A. Mey.) is a famous Chinese herbal medicine, it’s main medicinal ingredient is ginsenoside, especially rare ginsenosides have a stronger anti-tumor effect. The kinds and contents of ginsenoside are important standards of ginseng in the quality evaluation. Renshen Jianpi Pill, Renshen Guipi Pill, Renshen Zaizao Pill, Wuji Baifeng Pill are commonly used in the ginseng formulation, which have been used frequently in traditional Chinese medicine for thousands of years. But because of the lack of scientific detection methods and strict quality standards in traditional Chinese medicine preparation, there are still a low level of quality control, extensive pharmaceutical technology, unknown mechanism of action and other issues. Therefore, this study was to secondary evaluate quality and efficacy of ginseng preparations, and the development of new dosage form products. Firstly, a simple and accurate HPLC for determining the contents of 20 ginsenosides from ginseng and its preparation was established in the study. And the contents of 20 ginsenosides in 4 ginseng preparations were analyzed by HPLC; Secondly, the MTT assay in vitro for kinds of ginseng preparation of antitumor activity were screened; Third, the ginseng preparation was done according to the strongest anti-tumor activity, and then the compound granules capsules was made by optimizing optimum process; Finally, the compound granules capsules were studied for the purpose of the effect of anti-tumor in the human body, providing a theoretical basis for the secondary development of ginseng preparations and quality control. Experimental results were as follows:1. The method of high-performance liquid chromatography, which can determine 20 kinds of ginsenoside monomers(Rg1, Rh4, Rg2, Rg3, Rg5, Rf, Re, F1, F2, Rc, Rd, Rb1, Rb2, Rb3, Rh2, compound K, protopanaxatriol, 20(R)-Rh1, Rk3, protopanaxadiol) from ginseng and its preparation, was developed. The method was stable and accurate, which increased stronger anti-tumor activity of four kinds of rare ginsenosides and could be used as the detection standard of ginseng and its preparation with internal quality.2. The contents of ginsenosides of different manufacturers and the different ginseng preparations were different(P<0.05). Ginsenosides like Rg5、Rg3、Rk3、F2 in sun-dried ginseng were not found, while the content of Rh2 was increased. Instability effect of ginseng preparation was due to the quality of ingredients or processing problems. It’s important to establish the quality standard of ginseng preparations.3. There were differences of in vitro anti-tumor effects in different manufacturers of different formulations. Four kinds of ginseng preparations of MCF-7 had good antitumor activity, the best of which was Renshen Jianpi pill, and its IC50 concentration is 287.18 mu g/mL.4. The optimum reaction conditions were obtained by single factor experiments and response surface analysis, that is according to the proportion of Chinese pharmacopoeia, each herbs of Renshen Jianpi pill(with 60 mesh) were accurately weighed, added 10 times distilled water, soaked for 30 min, 100 ℃ heated and boiled for 90 min, decocted 2 times, filtrated. Concentrated the twice combined filtrate under reduced pressure to quantitatively. Then it was taken the ginseng into the filtrate, and the filtrate was fried with water bath by 100℃, dried it by 100℃, crushed and sieved into the Renshen Guipi powder, then made into Renshen Guipi particles.5. Renshen Guipi particles could inhibit tumor growth in mice tumor-burdened H22(P < 0.05), increase tumor mouse organ index, protect spleen organ, and promote tumor tissue mecrosis. It also could promote H22 tumor cell apoptosis(P < 0.05), improve immune cell activity and the rate of CD4+/CD8+. And it could increase the content of immunoglobulin IgG、IgE、IgM and TNF-α、IFN-γ、IL-2、IL-6、IL-12 in serum, reduce the content of TGF-β. Though 5-FU, which is positive drug, had a significant inhibitory effect(P < 0.05), it also reduced the immune organ index and spleen lymphocyte proliferation activity, with some side effects.