To Rhizoma Imperatae Anti-liver Cancer Substances Group Screening And In Vitro And In Vivo Anti-tumor Effect

PurposeRhizoma Imperatae is one of the major components of the Chinese ComPound Prescription treating liver cancer. This object is the national science and technology major subject 2009ZX09103-354 for drug research of the treatment of liver cancer. To?investigate the effects and preliminary mechanism of Rhizoma Imperatae on treating liver cancer and screen effective substance groups,a series of efficacy trials in vivo and vitro had been operated thorough study of effective parts of Chinese Prescription, make it possible to lead the development of Chinese medicine theory from compatibility of medicines to the compatibility of chemical components.Methods1 An optimal extractive method of aqueous extract and polysaccharide in Rhizoma Imperatae was fixed by HPLC and UV. The effective material group in Rhizoma Imperatae is purificated by macroporous resin. And the effectiveness was investigated by MTT method, in which the vitro cell growth inhibition and IC50 were choosed as the content index.2 To compare the inhibitive effects of aqueous extract and ethanol extract on proliferation of human hepatoma cell line by MTT colorimetric assay in vitro.?The inhibitory effects of aqueo -us extract and ethanol extract on tumor growth were observed by the models of implanted hepatoma 22(H22) in mice.3 The antiproliferative activities of growth curve of aqueous extract against human hepatoma cells line SMMC-7721 and Hep G2 were evaluated by MTT colorimetric assay in vitro.?The inhibitory effects of different doses of aqueous extract on tumor growth were observed by the models of implanted hepatoma 22(H22) in mice.4 The antiproliferative activities of growth curve of polysaccharide in Rhizoma Imperatae against human hepatoma cells line SMMC-7721 and Hep G2 were evaluated by MTT colorimetric assay in vitro. The inhibitory effects of different doses of polysaccharide in Rhizoma Imperatae on tumor growth were observed by the models of implanted hepatoma22(H22) in mice. 5 The immune function is evaluated by immune pharmacological tests (immune organ weight and organ index, synergism and reducing toxicity, splenic T, B lymphocyte transformation test, leucocyte medium 2 and interferon-γin ELISA kit).6 To determine quality control standards of the water extract and polysaccharide in Rhizoma Imperatae by HPLC and UV.7 The antiproliferative activities of Rhizoma Imperatae medicated serum against human hepatoma cells were evaluated by serum pharmacological in vitro. Elements of Rhizoma Imperatae serum were observed by microwave digestion and Inductively Coupled Plasma Mass Spectrometry (ICP-MS), the data were analyzed with the method of cluster analysis.Result1 The IC50 of water extract and alcohol extract of Rhizoma Imperatae on SMMC-7721 cells were 35.25 mg.mL-1 and 48.46mg.mL-1 respectively. The inhibitory effects of different doses of aqueous extract for the models of implanted hepatoma 22(H22) in mice is 33.68%, alcohol extract showed no inhibition effects. The results showed that the anti-liver cancer effect of water extract was significantly stronger than ethanol extract.2 The water extract of Rhizoma Imperatae inhibited the growth of HepG2 and SMMC-7721 cells in a dose-and time-dependent manner at dose of 2.5~40 mg.mL-1. The inhibitory rates of water extract on the growth of H22 at three dose were 20.68%,20.92%,41.83% respectively.3 The polysaccharide in Rhizoma Imperatae inhibited the growth of HepG2 and SMMC-7721 cells in a dose-and time-dependent manner at dose of 25~400μg.mL-1. The inhibitory rates of water extract on the growth of H22 at three dose were 35.95%,37.91%,32.03% respectively.4 The polysaccharide in Rhizoma Imperatae significantly increased the spleen index, produced efficacy enhaning and toxicity reducing combined with cyclophosphamide, and promote splenic T and B lymphocyte cells proliferation, improve IL-2 and INF-7 activity of peripheral blood.5 The optimal extraction process of water extract was with 16 times amount of water, reflux for 3 times, 1h. The optimal extraction process of Rhizoma Imperatae polysaccharide was with 10 times amount of ad water, 100℃extract 3 times, 2.5 h. Filter out Macroporous resin and the elution solvent concentration: AB-8 macroporous resin 70% ethanol elution.6 The medicated serum of water extract and polysaccharide inhibited the proliferation of the Hep G2 and SMMC-7721 cells, compared to the blank group, the difference was significan (P <0.01).7 The data of water extract were analyzed with the method of cluster analysis, treated group and blank group were divided into a group by Ca,Cu, Zn,Se. The data of polysaccharide in Rhizoma Imperatae were analyzed with the method of cluster analysis, treated group and blank group were divided into a group by Na,Ca,Cu,Zn.Conclusion1 Water extract was the effective substance groups in Rhizoma Imperatae, the polysaccharide in Rhizoma Imperatae was one of the major components of effective substance groups.2 Efficacy trials in vivo and vitro proved that the aqueous extract of Rhizoma Imperatae possed an anti-hepatoma effect.3 Efficacy trials in vivo and vitro proved that the polysaccharide in Rhizoma Imperatae possed an anti-hepatoma effect.4 The immunomodulatory function of polysaccharide Rhizoma Imperatae was initially recognized.5 The anti-hepatoma effect of the water extract and polysaccharide in Rhizoma Imperatae was proved again by the serum pharmacology study.6 Ca,Cu,Zn may be related to the occurrence and development of cancer.