Experimental Study On The Effect Of Polysaccharide Of Radix Glehniae On Immune Function Of Mice And RAW 264.7 Cells

Objective:Radix Glehniae,as a dual-purpose medicine,has rich active ingredients and obvious pharmacological effects,so it has a good prospect of development and utilization.There are many complex components in Radix Glehniae,among which polysaccharides account for the largest proportion,and they are the main components to enhance and regulate immune function.In this paper,the polysaccharide of Radix Glehniae was used as the research object to explore its influence on the immune function,so as to give new vitality to the further development and utilization of health food with immune function of polysaccharide of Radix Glehniae,and to provide theoretical basis for the application of authentic medicinal materials in Changbai Mountain.Method:In vitro cell experiments:①In this experiment,the polysaccharide was extracted by ultrasonic extraction,hot water extraction and cellulase extraction,and the protein was removed by Sevage.The phenol-sulfuric acid method was used to measure the total sugar content of the polysaccharide of Radix Glehniae,and its precision and repeatability were investigated.②By setting the concentrations of 37.5,75,150 and 300 μg/mL,the polysaccharide was co cultured with mouse macrophages(RAW 264.7),using ELISA method to detect the co-cultured cell supernatant changes in the content of immune factors interleukin-6(IL-6)and tumor necrosis factor(TNF-α);MTT colorimetry was used to determine the effect of polysaccharide of Radix Glehniae on the proliferation of macrophages(RAW 264.7);Griess was used method to determine the change of nitric oxide(NO)in the cell supernatant.③The polysaccharide of Radix Glehniae was induced by CON A to induce mouse spleen cells,the content of IL-2 and IFN-γ in spleen cells of mice stimulated was studied.In vivo animal experiments:BALB/C mice were divided into low(200 mg/mL),medium(400 mg/mL),and high(800 mg/mL)polysaccharide groups and blank control groups.The polysaccharide group was fed by gavage according to the proportion of body weight,and the blank control group was fed with normal saline according to the proportion of body weight for 28 days.Detect the weight change,thymus and spleen index of mice;LDH method was used to detect the effect of co culture of mouse spleen cells and target cells YAC-1 on NK natural killer cell activity;CCK-8 method was used to detect the proliferation of T and B lymphocytes in the spleen of mice;ELISA was used to detect changes in cytokines in mouse serum;Flow cytometry was used to detect the distribution of lymphoid subgroups in mouse spleen cells.Result:In vitro cell experiments:①The extraction rate of polysaccharide from Radix Glehniae was as follows:ultrasonic extraction method(14.89%)<hot water extraction method(23.93%)<cellulase extraction method(24.22%),polysaccharide content:cellulase extraction method(58.05%)<ultrasonic extraction method(73.25%)<hot water extraction method(75.48%).Therefore,hot water extraction is the best method.The precision R of the extraction methods are lower than 5%,and the repeatability R are all below 6%.The results show that the hot water extraction method is accurate and reliable for the extraction of polysaccharides from Radix Glehniae.② In the range of 37.5-300 μg/mL,the content of IL-6、TNF-α and the secretion level of NO were significantly increased compared with the blank control group.The polysaccharide from Radix Glehniae promoted the content of IL-2 and IFN-γ secreted by spleen cells induced by CON A,and the concentration increased in a dose-dependent manner.In vivo animal experiments:The polysaccharide of Radix Glehniae group has no effect on the body weight of mice,and has a significant increase in thymus and spleen indexes of mice,with statistical differences.The polysaccharide group significantly increased the NK cell activity and the proliferation of T and B lymphocytes with statistical differences;the polysaccharide of Radix Glehniae group could increase the cytokines IL-2,IL-6,TNF-α and IFN-y in mouse serum within a certain range.The content of serotonin was statistically significant,which has statistical significance;The results of lymphocyte subsets distribution experiment showed that the polysaccharide group could significantly increase the percentage of CD3+,CD3+CD4+,and CD3-CD19+cells,reduce the percentage of CD3+CD8+,and increase the ratio of CD4+/CD8+in the spleen of mice.Therefore,all results indicates that the polysaccharide of Radix Glehniae can regulate the distribution of lymphocyte subsets.Conclusion:The polysaccharide of Radix Glehniae can improve the immune function of the body by increasing the proliferation and differentiation of macrophages and lymphocytes.It can increase the activity of T,B cells,NK cells,etc.of mice by acting on lymphoid subgroups,and enhance the body’s Non-specific and specific immune functions.