Study Of Auricularia Auricula Polysaccharides’ Anti-melanoma And Its Related Mechanism

Melanoma is a type of malignant tumor originated from melanocytic abnormalities.It is the most malignant tumor in skin tumors and is prone to metastasis.At present,the commonly used methods of treatment include chemotherapy,but chemotherapy drugs have certain toxic and side effects,which do not achieve good therapeutic effect.As a natural active substance,Auricularia auricula polysaccharide has been proved to have a wide range of physiological activity,including anti-tumor effect.In this thesis,we mainly explore the effect and possible mechanism of Auricularia auricula polysaccharides on the anti-tumor of melanoma by establishing the model of melanoma in vivo and the in vitro experiment targeting B16 cells,which will provide theoretical basis and train of thought for the clinical research of melanoma drugs.Objective To investigate the anti-melanoma effect of Auricularia auricula polysaccharide,as well as to prove the related mechanisms combined with cytological experiments.Methods Acute toxicity experimental model was established to observe the acute toxicity of Auricularia auricula polysaccharides;The melanoma model was established,and after Auricularia auricula polysaccharides administrated,the survival rate of mice was observed and the tumor size was measured,then the anti-tumor effect of Auricularia auricula polysaccharide was preliminarily determined.Next,the expression of apoptotic genes and cytokines in tumor tissues were detected by immunofluorescence technique.The activity of NK cells and CTL cells in spleen was measured and the expression of related factors of killer cells was detected by flow cytometry,which the possible anti-tumor mechanism of Auricularia auricula polysaccharides was detected.The cell proliferation and apoptosis rate were determined by MTT and Annexin V/ PI staining,which the direct anti-tumor effects of Auricularia auricula polysaccharides on melanoma were detected.Results 1.Auricularia auricula polysaccharide is basically non-toxic or weak toxicity.Intragastric administration of high dose of Auricularia auricula polysaccharide in a day,the mice all survived and the mental state and activities had no significant difference compared with pre-administration.At the same time,the food intake were increased,the growth of body weight in the dose-group of 200mg/kg.BW and 400mg/kg.BW was significantly decreased than that of nomal saline group(P<0.05),the content of RBC,HGB and PLT in mice were lightly changed by administration of Auricularia auricula polysaccharide.2.Auricularia auricula polysaccharide has antitumor effect on melanoma in vivo The melanoma model was established of,and after administration of Auricularia auricula polysaccharide,the survival time of mice increased significantly,the tumor size and tumor weight decreased,the expression of apoptotic genes and cytokines in tumor tissues was obsviouly increased.3.The immune cells of melanoma mice activated by Auricularia auricula polysaccharide After the melanoma mice were administrated of Auricularia auricula polysaccharide,the number of white blood cells was increased,the activity of NK cells and CTL was enhanced.Auricularia auricula polysaccharide promoted the secretion of perforin and granzyme B,which was produced by NK cells and CTL.The secretion of IFN-g by CD4+ T cells and CD8+ T cells was also increased.4.Effect of Auricularia auricula polysaccharide on proliferation and apoptosis of B16 cells B16 cells were treated with different concentrations of Auricularia polysaccharide,the inherent morphology of B16 cells was changed,the cell healing rate of sratches was decreased and the activity of B16 cells was in a dose-dependent manner.The activity of B16 cells had the best effect and the apoptosis rate were increased after treated with Auricularia auricula polysaccharide for 24 h,which IC50 was 21.53μg/ml.Conclusion Auricularia auricula polysaccharide has anti-tumor effect on melanoma,its mechanism may be directly related to the apoptosis of B16 cells induced by Auricularia auricula polysaccharides,or by activating immune cells,and to promote the secretion of perforin and Granzyme B of NK cells and CD8+T cells,perforin and Granzyme B could directly act on tumor cells.