Preparation Process Of Sanghuang Polysaccharide Granules And Its Anti-inflammatory Mechanism On Ulcerative Colitis In C57BL/6 Mice

Objective: UC is a chronic continuous diffuse inflammatory disease whose pathogenesis is still unclear.In recent years,the incidence in China has continued to increase.It mainly affects the colonic mucosa,and the lesions mostly occur in the distal colon.UC is a long-lasting and prone to recurrent disease,which is a global disease.Sanghuang polysaccharide,as a non-toxic chemical component,has a very prominent anti-inflammatory effect,and it will be prepared into a health food drug,which is beneficial to long-term use by patients with ulcerative colitis to relieve symptoms.The purpose of this project is to verify the therapeutic and protective effect of Sanghuang polysaccharides on ulcerative colitis;to study the preparation process of Sanghuang polysaccharides granules,and to lay the foundation for its dosage form in the treatment of ulcerative colitis.Method: Sanghuang polysaccharide was extracted by hot water extraction,and the polysaccharide extraction method was optimized by single factor and response surface method.After a series of purification,the refined Sanghuang polysaccharide was obtained.In order to expand the application of Sanghuang polysaccharide,research The preparation process of Sanghuang polysaccharide granules was studied,and the preparation method of granules was preliminarily determined.Whether Sanghuang polysaccharide has protective effect on ulcerative colitis was verified by in vitro cell experiment and mouse experiment.Cell experiment: RAW264.7 cells were selected,LPS was used as the drug to induce inflammation,and a blank group,a model group,and a high-medium-low-dose administration group were set up.Mice experiment: Select C57L/4B mice,20—22g each,and divide them into blank group,positive drug group,model group,high-low dose administration group,10 mice in each group,randomly assigned,DSS is used as an inflammation-inducing drug,3% m/v drinking water was administered for seven days,and water was freely available.From the first day,the model group and the blank group were given distilled water,and the positive drug group and the high and low dose administration groups were given corresponding volumes of drugs.Results: The results of single factor experiment and response surface method optimization showed that the extraction temperature of Sanghuang polysaccharide was 75℃,the extraction time was 2h,and the ratio of solid to liquid was 1:16g/ml.The yield is the highest under the conditions;the results of the granule preparation process show that soluble starch and lactose are selected as auxiliary materials,80% ethanol is used as a wetting agent,the ratio of mixed auxiliary materials is starch: lactose(3:1),and the ratio of raw materials and auxiliary materials is 1: 2.Use this condition as the preliminary preparation process of Sanghuang polysaccharide granules;the results of cell experiments show that the concentration of Sanghuang polysaccharide is 7.8125-31.25ug/ml,the concentration of LPS is 500ng/ml,and the Elisa kit detects the inflammatory factor IL-6,TNF-α,the concentration of the administration group was significantly lower than that of the model group,and the expression of IL-6,IL-1β,TNF-α m RNA was detected by pcr,and the m RNA scalar amount of the administration group was significantly lower than that of the model group;mouse experiments showed,Sanghuang polysaccharide dosage was 50mg/kg,100mg/kg,3%DSS drinking water was used to build the model,Elisa kit was used to detect inflammatory factors IL-6,IL-1β,TNF-α,the concentration of the administration group was significantly lower than that of the model group,pcr detected the m RNA expression of IL-6,IL-1β,TNF-α,and the m RNA scalar quantity of the drug administration group was significantly lower than that of the model group;the results of Western blot experiments showed that Sanghuang polysaccharides were activated in both cells and mice.NF-κB,AKT,and MAPK pathways regulate related proteins.Conclusion: Through the optimization of the extraction process of Sanghuang polysaccharides,it is determined that the extraction temperature is 75°C,the extraction time is 2h,and the ratio of solid to liquid is 1:16mg/ml.1.Insufficient extraction in a short period of time,too little water will cause the polysaccharides to fail to dissolve completely;in the process investigation of granules,due to the properties of Sanghuang polysaccharides,too much polysaccharides will make the granules viscous and cannot be produced in the ratio of raw and auxiliary materials.Granules,better formability of mixed excipients;in the pharmacological activity experiment,the anti-inflammatory effect of Sanghuang was studied,and the results of in vivo and in vitro experiments showed that it had certain anti-inflammatory activity and activated NF-κB,AKT,MAPK pathway regulates related proteins,and preliminarily determines the anti-inflammatory mechanism of Sanghuang polysaccharide.