Nanoformulations Of Ganoderma Lucidum Polysaccharide Enhanced The Antitumor Effect Of Doxorubicin And Reduced The Side Effects

Objective: To investigate the immunomodulatory activity of nanoformulations of Ganoderma lucidum polysaccharide(GLP)and their antitumor effects with.doxorubicin Methods:1.Purification of Ganoderma lucidum polysaccharide: After deproteinization and depigmentation of Ganoderma lucidum crude polysaccharide,the polysaccharide is separated and purified by anion exchange column and gel column to collect uniform molecular weight and negative polysaccharides.2.Nanometerization of Ganoderma polysaccharide: Use The chemotherapeutic drug doxorubicin(Dox)and Ganoderma lucidum polysaccharides are used to prepare polysaccharide-doxorubicin nanoparticles(Dox@GLP)in one step.3.Various characteristics of nanoparticles: hydrated particle size(DLS)and dispersion degree(PDI)of nanoparticles,Zeta potential,transmission electron microscope and fluorescence emission spectra.4.To study the toxicity of nanoparticles to fine 4T1-luc,NIH/3T3 and DC cells,study the uptake of nanoparticles by 4T1-luc,and observe the incorporation of nanoparticles,To study the effect of nanoparticles on the expression of histocompatibility complex II(MHCII)and costimulatory molecules(CD40,CD86)on the surface of DC.5.To study the distribution and metabolism of nanoparticles in the body.6.To establish subcutaneous mammary glands in mice Carcinoma(4T1)model,to evaluate the anti-tumor effect of Dox@GLP in vivo,and to investigate the immune activation effect of Dox@GLP on tumor-bearing mice.Results:The prepared Ganoderma lucidum polysaccharide nanoparticles have suitable hydrated particle size,uniform particle size,and stability.Transmission electron microscopy,dynamic light scattering,and ultraviolet absorption spectroscopy fluorescence spectroscopy verify the success of the preparation of the nanoparticles,and also prove that the outer layer of the nanoparticles is hydrophilic.The inner layer of Ganoderma lucidum polysaccharide is hydrophobic doxorubicin.In vitro Ganoderma lucidum polysaccharide nanoparticles can inhibit the proliferation of 4T1-luc,and its inhibitory effect on primary DCs is reduced compared with that.Ganoderma lucidum polysaccharide nanoparticles can significantly up-regulate the expression of CD40,CD86 and MHCII in mouse bone marrow-derived DCs,mature the DCs and improve the ability of DCs to present antigens.The distribution in the body confirmed that the Ganoderma lucidum polysaccharide nanoparticles are enriched in the tumor,and the nanoparticles are mainly metabolized by the liver and kidney.Ganoderma lucidum polysaccharide nanoparticles can also stimulate DCs in tumor-bearing mice,increase the ratio of CD8+T in lymph nodes and tumor immune microenvironment,and enhance the immune memory ability of mice.Conclusion:The results of the study show that Ganoderma lucidum polysaccharide nanoparticles(Dox@GLP)can directly kill tumor cells,and at the same time,it can achieve specific cellular immunity by activating DC and produce anti-tumor effects.Doxorubicin can cause the immunogenic death of tumor cells,expose tumor-associated antigens,and recruit DC cells to present antigens.The polysaccharide nanoparticles can stimulate DC maturation and improve its antigen presentation ability,thereby enhancing the anti-tumor immune effect caused by chemotherapy.This research provides new research ideas for the application of modern dosage forms of traditional Chinese medicine polysaccharides in the treatment of tumors,and is conducive to the modern application of Chinese medicine.