The Research On Preparation Technolony And Evalution Of Antitumor Activity From The Orange Peel Extraction

This paper uses aituri of Hainan as material to obtain extract from aituri peel with NaOH solution.In order to obtain maximum extraction rate,the optimal extraction conditions should be explored.By single factor and orthogonal test,we get the optimum extraction condition:the ratio of sample to solution is 1:30,the concentration of NaOH is 0.6 mol/L,extracting time is 10 h,extracting temperature is 60?.Under the condition,the extraction yield reaches to 13.89%.According to FT-IR and UV spectra,the ingredient of aituri peel can be analysis preliminarily.Through a series of qualitative and quantitative experimental,the aituri peel extraction contains are polysaccharide 7.5%?including uronic acid 3.94%?,protein 11.45%,polyphenols 4.93%,12.83%sulfuric acid group and hesperidin were 8.72%,11.35%flavonoids,yellow pigment 9.83%,pectin 2.71%,ash content 0.6%.In guarantee the minimum lavage quantity to reach highest inhibitory rate,experiment we acquire the optimal lavage amount is 50 mg/kg · d by animals experiments.During the lavage,the mice have good mental state and no abnormal reaction happen.The luster fur and normal movement of mice illustrate non-toxic of the extraction.In order to explore its anti-tumor effect,animal and cell experiment cried out in vivo and vitro,the experiment in vitro simulated human body environment as far as possible.The result shows the extraction can not directly affect tumor cells but by protecting the immune organs in vivo then improve the level of the immune implement antitumor effect indirectly.In vivo experiment,we look at the aspects of humoral immunity and cellular immunity to study the anti-tumor effect,and through the test in mice blood and immune organ index to explore its antitumor mechanism.This paper focuses on the improvement of immunity by protecting immune organs.Also,we explore the mechanism of the extraction against tumor,which provides wider reference for the tumor treatment in the future.This experiment use H₂₂ liver cancer cells to establish mice model.Through the body experiments,we explored the antitumor effect of aituri peel extraction with the dosage of 50 mg/kg·d.In vivo experiments,it shows that the cure rate of the extraction in tumor-burdened mice reaches 60%,which indicate theextraction can effectively inhibit the growth of solid tumor.By measuring the immune organ index of mice and observing the paraffin section of the organization,we found the extraction is good to protect the immune organs.By detecting the spleen lymphocytes in mice under ConA and LPS induced proliferation,we found that the extraction can strengthen the proliferation activity of mice spleen T lymphocytes and B lymphocytes.From the test of NK cell killing ability and the level of phagocytosis of macrophage cell,we come to the conclusion that medicine group can significantly increase the killing activity of NK cells,activate the mice macrophage activity,improve their ability of phagocytosis and enhance the immune function of mice.Lymphocyte subsets results showed that the amount of T lymphocyte increase obviously by gavaged with extraction.Among those tests,gavage group have a high quantity of T lymphocytes with increasing number of CD4+T cells.The detection results of cytokines in mice serum IL-2?IFN-??TNF-? and Elisa experiments also prove that the extraction can regulate immune factors,and improve the ability of the body's immune function.MTT in vitro experiments show that different concentrations of extraction can not effectively play its role in inhibiting tumor,which suggesting that the inhibition effect of aituri peel extraction on tumor suppression is indirectly by improving immunity.