Study On The Immunological Activity Of Ligusticum Chuanxiong Polysaccharide And The Preparation Process Of Its Effervescent Tablets

Polysaccharide is a recognized natural immunomodulator,widely distributed in animals,plants and microorganisms in nature.Ligusticum chuanxiong has a high content of polysaccharides,but they are often discarded,causing a lot of waste.The purpose of this study is to study the neglected parts of Ligusticum chuanxiong polysaccharides,to screen the best extraction process,to explore its immune activity,and to develop a clinically suitable dosage form.In order to expand the medicinal range of Ligusticum chuanxiong,increase the comprehensive utilization rate of Ligusticum chuanxiong,and reduce the waste of medicinal materials.The specific research methods and results are as follows:1.Preparation process of crude polysaccharide of Ligusticum chuanxiongTaking the yield of crude polysaccharides of Ligusticum chuanxiong as the index,water extraction and alcohol precipitation method were used to prepare Ligusticum chuanxiong polysaccharides.The extraction technology and alcohol precipitation technology of Ligusticum chuanxiong polysaccharides were investigated,and the optimal preparation technology was determined as follows: After 24 mesh sieve,Ligusticum chuanxiong powder was refluxed and extracted twice at 90℃,each time for 2 hours.The supernatant was combined and concentrated to 1.5g/m L(original medicinal materials).95% ethanol was added until the final alcohol concentration reached 80%.2.Quality analysis of Ligusticum chuanxiong polysaccharideThe protein in the crude polysaccharide of Ligusticum chuanxiong was removed by trichloroacetic acid-n-butanol method to obtain Ligusticum chuanxiong polysaccharide.The analysis of Ligusticum chuanxiong polysaccharide showed that it was a yellowbrown powder,slightly soluble in water,with protein content of 0.86%,moisture content of 2.97%,polysaccharide content of 80.15% in terms of glucose,and the results of infrared spectroscopy showed that it was an α-glycosidic bonded pyranocyclic polysaccharide compound,and the results of HPLC showed that Ligusticum chuanxiong polysaccharide contained mannose,galacturonic acid,glucuronide,galactose and arabinose in the molar ratio of 0.022:0.435:1:0.145:0.125,and the GPC results showed that the molecular weight distribution was mainly concentrated in the range of 45000.0 ～220000.0 g/mol.3.Study on immune activity of Ligusticum chuanxiong polysaccharideKM mice were used as experimental animals and levamisole hydrochloride(50 mg/kg·bw)was used as positive drug.An immunosuppression model was established in mice by intraperitoneal injection of cyclophosphamide(50 mg/kg·bw),and low(50 mg/kg·bw),medium(100 mg/kg·bw)and high(200 mg/kg·bw)doses of Ligusticum chuanxiong polysaccharide were given by gavage.The immune organ index was measured to observe the immune organ damage,the phagocytic ability of macrophages was determined by carbon particle contouring assay,the antibody production level was reacted by serum hemolysin assay,and the serum levels of IL-2,IL-6,IL-10 and IFN-γ were detected by Elisa kit to evaluate the immune activity of Ligusticum chuanxiong polysaccharide.The experimental results showed that Ligusticum chuanxiong polysaccharide improved all of the above immune indexes.The results of 16 S macrogenome sequencing assay of mouse intestinal flora showed that Ligusticum chuanxiong polysaccharide could significantly improve the dysregulation of intestinal flora and increase the diversity of intestinal flora in immunosuppressed mice.The 16 S functional gene prediction analysis was performed to predict the gene function of the samples,and the correlation between the flora and the immune indexes with the flora was explored.4.Study on the Preparation Technology of Ligusticum Chuanxiong Polysaccharide Effervescent TabletsTaking the cumulative dissolution as the index,the best preparation process of Ligusticum chuanxiong polysaccharide solid dispersion was determined as follows: the solid dispersion was prepared by melting method,PEG6000 was used as carrier material,and the drug-load ratio was 1:5.The solid dispersion of Ligusticum chuanxiong polysaccharide was characterized to judge whether the preparation was successful or not.Then Ligusticum chuanxiong polysaccharide solid dispersion effervescent tablets were prepared by using Ligusticum chuanxiong polysaccharide solid dispersion as intermediate.The best preparation process of effervescent tablets was determined as follows: using acid-base separation granulation method,using mannitol as filler,sodium bicarbonate as alkali source,citric acid as acid source,acid-base mass ratio as 2:1,disintegrating agent50%,Ligusticum chuanxiong polysaccharide solid dispersion 30%,anhydrous ethanol as non-aqueous binder,3% sodium chloride as lubricant,2% aspartame as sweetener.The quality inspection and preliminary stability test of Ligusticum chuanxiong polysaccharide effervescent tablets were in accordance with the regulations.