The Research On Anti-tumor Activities Of Total Polysaccharides From Zanthoxylum Dissitum Pericarp

Zanthoxylum dissitum belongs to the family of Rutaceae. Its root, stem bark andleaf can be used as folk medicine used to treat stasis to relieve pain, detoxificationswelling, joint pains, injuries from falls, etc. in the civilian society of China. In order toimprove the exploiting and using of Zanthoxylum dissitum, the investigations on theanti-tumor activity of have been carried out in this paper.The chemical constituents of Zanthoxylum dissitum have been testedsystematically by means of pre-test. It found that it contains alkaloids, saponins,flavoniods and glycosides, cardiac glycosides, anthraquinones, saccharides andglycosides, volatile oils, grease, organic acids, amino acid, proteins; there is no phenolsor tannin, coumarin or terpenoid, steroids or terpenoids.And then the bioactivities of the polysaccharides, total alkaloids and total saponinhave been studied:（1） their antimicrobial activities in vitro were evaluated by filterpaper method. The results showed that the total saponin had no inhibitory effects to thefour bacteria （Klebsiella pneumonia, Streptococcus uberis, Staphylococcus aureus,Eschercihia Coli）, while the polysaccharides showed different inhibitory effects to thefour bacteria, and it showed the inhibitory effects to Staphylococcus aureus andEschercihia Coli when polysaccharides was at the concentration of10mg/mL. Thetotal alkaloids showed a little inhibitory effects to the four bacteria when it was at theconcentration of10mg/mL.（2） MTT colorimetric assay was adopted to test theiranti-tumor activities to human hepatoma cell lines Hep-G2. The results showed that thetotal saponin of Zanthoxylum dissitum exhibited no significant inhibitory effects onhuman liver tumor cells. The polysaccharides and total alkaloids had significantinhibitory effects on human liver tumor cells.To optimize the extraction of polysaccharides from Zanthoxylum dissitumPericarp by the use of response surface methodology, experiment factors and levelswere first selected on the ground of single factor test. Along with the central compositeexperimental design principles, the response surface methodology with three factorsand three levels was adopted in search of multiple quadratic linear regression analysis.Response surface and contour were finally chosen according to the extraction rate andthe response value, respectively. The optimum extraction parameters of thepolysaccharides from Zanthoxylum dissitum Pericarp were concluded as follows:extraction temperature was94℃, liquid-to-solid ratio was82︰1, extraction time was 146min, extracted twice. Under these optimized conditions, the extraction rate ofpolysaccharides is up to4.99%. Multivariate regression analysis showed that the modelof extraction temperature, liquid-to-solid, extraction time and extraction rate ofpolysaccharides could be forecasted by applying the model.In order to observe the anti-tumor activities of different dose of the totalpolysaccharides from Zanthoxylum dissitum pericarp in vitro and vivo, its anti-tumoractivity in vitro and in vivo was evaluated by MTT colorimetric assay and H22cells-bearing mice models. The results showed that the total polysaccharides fromZanthoxylum dissitum pericarp significantly inhibited proliferation of the humancervical carcinoma cell Hela and the human hepatoma cell lines SMMC-7721, Hep-G2at the concentration of62.5¹000μg·mL^-1, and the IC50value on these two tumor cellswere460.9μg·mL^-1,425.6μg·mL^-1and570.4μg·mL^-1, respectively. In vivo, high (1.5g·kg^-1), middle (0.75g·kg^-1) and low (0.375g·kg^-1) dose total polysaccharides fromZanthoxylum dissitum pericarp also significantly suppressed the tumor growth withtumor inhibition rate of41.55%,34.96%and33.71%respectively. The Slice throughthe pathological observation of tumor tissue also indicated that it has significantanti-tumor activity. Meanwhile, the results indicated the total polysaccharides fromZanthoxylum dissitum pericarp can display certain strengthen action to the immuneorgan of tumor-bearing and did not have other side effects to tissues of mice.