The Effect Of Grifola Frondosa Polysaccharides On The Pathological Changes Of The Small Intestine And The Expression Of TNF-? In Mice With Intestinal Infection Of Candida Albicans With Spleen Deficiency

Purpose:To explore the mechanism of traditional Chinese tonifying Qi therapy on the immune system,the model of Candida albicans infection in spleen deficiency mouse is established.The intervention effect of Grifola frondosa polysaccharide on the model mouse is discussed.Reveal the mechanisms of Grifola frondosa anti-infection effect,to provide a reliable experimental basis for the treatment of candidiasis albicans with Chinese herb that tonifying Qi and invigorating spleen.Material and methods:100 SPF Kunming mouse weighing 18-22g,half male and female.They were divided into normal control group(Group A),spleen deficiency model group(Group B),spleen deficiency+Candida albicans group(Group C),spleen deficiency+Candida albicans+Fluconazole group(Group D),and spleen deficiency+Candida albicans+Grifola frondosa polysaccharide group(Group E).Spleen deficiency model isestablished by swimming to the limit of strength and feeding cabbage and lard for two week.Then infected model is established by feeding Candida albicans with a concentration of 2×10~8CFU/mL.From the next day the mice in Group D and E are fed with drug once a day for one week.The mice in Group D are fed with fluconazole(concentration:26mg/kg),and in Group E are fed with grifola frondosa polysaccharide(concentration:10mg/kg).During this period,the general state change of mouse in each group was observed.After drug therapy,the mice were weighed and the feces was collected,the determination of the amount of bacteria in the manure was calculated,the jejunum tissues of each group were collected and made into pathological section,pathological changes of intestinal mucosa was observed by optical microscope.The expression of TNF-?in intestinal tissue were detected by RT-PCR.Results:1.The Changes of weight and general state in mouse.After two weeks in experiment,the spleen deficiency model has established.Except group A,the mouse in each group has shown typical symptoms of spleen deficiency such as weight loss,the ncrease of defecation,loose stool and so on.Comapred with group A and group B,the mouse in group C eat less and the weight were less than other groups,general state is worse than group A.After one week of drug therapy,there are no difference in weight between group D,E and A.The weight of group B and C was significantly less than groupA,group B and group C.The loss of weight in group C was more particularly significant.2.Determination of viable bacteria in fecesThe 7th of drug therapy,there was no significant statistical difference in fecal viable bacteria among groupA,group D and group E.The viable count of fecal bacteria in group B and C was significantly higher than group A.There was no significant difference in the count of fecal viable bacteria between group B and group D and E,the count of group C was more higher than the other groups.3.Morphological changes of small intestinal mucosa in each group.The intestinal mucosa of mice in group A are intact,intestinal villi arranged neatly,the thickness of muscular layer of mucosa is uniform and moderate.Compared to group A,pathological changes in group B are not obvious,occasionally the villi of the small intestine are sparse and irregularly arranged.The mouse in group C showed the loss of villi,the arrangement of intestinal glands and villi is irregular,inflammatory cell infiltration is seen in lamina propria and submucosa.After treatment,the pathological state of group D and E is gradually recovered,the arrangement of intestinal glands and villi is regular under optical microscope.Inflammatory cells are significantly less than group C.But it was significantly different compared to the group A.4.Detection of TNF-?expression in intestine tissue by RT-PCR.At the end of the experiment,the results of RT-PCR showd that the expression of TNF-?mRNA did not differ between group A and B.The highest expression level was found in group C among the other four groups.There was no statistical difference between D and E groups,and the expression of TNF-?mRNA in these group was significantly higher than in group A and B.Conclusions:1.Grifola frondosa polysaccharide can increase the weight of spleen deficiency mouse infected with Candida albicans,and improve their general conditions.2.Grifola frondosa polysaccharide can decrease fecal viable count of spleen deficiency mouse infected with Candida albicans.3.Grifola frondosa polysaccharide can improve the pathological state of intestinal mucosa in spleen deficiency mouse infected with Candida albicans.4.Grifola frondosa polysaccharide can decrease the expression of TNF-?mRNA of spleen deficiency mouse infected with Candida albicans.