Effect And Its Mechanisms Of Armillariella Oral Solution On 5-Fluorouracil-Induced Intestinal Mucosal Injury In Mice

Objective To investigate the effect of Armillariella Oral Solution on intestinal mucosal injury in mice after 5-FU(5-fluorouracil)chemotherapy and its related mechanism.Method A total of sixty C57BL/6 mice were randomly divided into six groups:control group,model group,Armillariella Oral Solution group at dose of 1 ml/kg,5ml/kg,10 ml/kg,positive control group(live combined bifidobacterium and lactobacillus tablets,CBL,450 mg/kg).All mice,except for control group,received5-FU(50 mg/kg,once daily,intraperitoneal injection)for 7 days.Drug treatment group received AOS,CBL(via oral gavage)once daily for continuous 14 days before 5-FU treatment for a week,control group received saline solution instead of 5-FU.During the course of chemotherapy,the mice were observed for bite and sup,event,pelage,body weight,and the mice were sacrificed 24 hours after the end of chemotherapy.Intestinal tissue was used to examine intestinal mucosal injury by hematoxylin-eosin(HE)staining;fluorescein isothiocyanate-dextran(FITC-Dextran)tracer method was used to detect intestinal mucosal permeability;enzyme-linked immunosorbent assay(ELISA)was used to detect endotoxin in plasma;bacterial plate count was used to detect intestinal bacterial translocation;immunohistochemistry(IHC)was used to examine the expression of intestinal tight junction protein zonula occluden 1(ZO-1),occluded protein(Occludin),intestinal crypt cell proliferation antigen(Ki-67),cysteinyl aspartate specific proteinase 3(Caspase-3),leucinerich repeat containing G protein-coupled receptor 5(Lgr5);tdt-mediated dUTP nick-end labeling(TUNEL)staining for detection of intestinal crypt cells apoptosis.Result 1.Compared with the control group,the HE staining of the model groupindicated that the intestinal mucosal injury of the mice was obvious,the villus atrophy was merged,the crypts were deepened or disappeared,and a mass of inflammatory cells infiltrated;the plasma concentration of FITC-Dextran,endotoxin and the intestinal bacterial translocation rate in the model group were remarkablely increased(P<0.05);immunohistochemistry demonstrated that the expression of tight junction protein ZO-1,Occludin,and the expression of Ki-67 and Lgr5 in the model group were significantly decreased(P<0.01),while the expression of Caspase-3 was increased(P<0.01);TUNEL staining showed that the apoptotic cells in the intestinal crypt of the model group were increased(P<0.01);2.Compared with the model group,the AOS medium and high dose mice intestinal mucosal damage was alleviated,villus and crypt were partially restored,and a small amount of inflammatory cells infiltrated;the plasma levels of FITC-Dextran,endotoxin and the intestinal bacterial translocation rate were significantly significantly decreased(P<0.05);immunohistochemistry indicated that the expression of tight junction protein ZO-1,Occludin,and the expression of Ki-67 and Lgr5 were significantly augmented(P<0.01),while the expression of Caspase-3 was reduced(P<0.01);TUNEL staining exhibited that the apoptotic cells in the intestinal crypt were lowered(P<0.01).Conclusion 1.Armillariella Oral Solution alleviate intestinal mucosal damage in mice after 5-FU chemotherapy,which may be related to lower the permeability of intestinal mucosal,restrain endotoxin and bacterial translocation;2.Armillariella Oral Solution alleviate intestinal mucosal damage in mice after 5-FU chemotherapy,which may be related to up-regulate the expression of Lgr5 in intestinal crypt,inhibit apoptosis and promove recovery of intestinal crypt cells.