Objective This study aimed to develop a type of Ganoderma lucidum(G.lucidum)-probiotic fermentation broth that can effectively improve the intestinal mucosal barrier function of mice with ceftriaxone-induced intestina...Objective This study aimed to develop a type of Ganoderma lucidum(G.lucidum)-probiotic fermentation broth that can effectively improve the intestinal mucosal barrier function of mice with ceftriaxone-induced intestinal dysbiosis.Methods By means of absorbance of optical density(OD)value and phenol-concentrated sulfuric acid measurement of polysaccharide content,the probiotic species can grow on the medium of G.lucidum were screened out,and the concentration of the medium of G.lucidum was determined,and the fermentation broth was prepared for subsequent experiments.Thirty-two SPF grade male BALB/c mice were randomly divided into four groups(eight mice in each group),namely control group(CON),intestinal mucosal barrier damage model group(CS),fermentation broth intervention group(FT)and G.lucidum medium intervention group(GL),respectively.The intestinal dysregulation model was induced by intra-gastric administration of 0.2 mL ceftriaxone sodium(twice a day for seven consecutive days).From day 8,the FT group and GL group were gavage with 0.2 mL fermentation broth and G.lucidum medium,respectively.On day 15,all mice were sacrificed.To draw the weight curve and measure the cecal index;pathological examination of colon tissues with HE staining;serum levels of LPS,IL-10,TNF and IL-6 were detected by ELISA.Flow cytometry was used to analyze the changes of CD3+T cells,CD4+T cells,CD8+T cells and macrophages in spleen.16S rRNA sequencing was performed to detect the intestinal microbiota structure of mice.Results Bacillus subtilis can decompose and utilize G.lucidum fruiting body medium,and the suitable concentration of G.lucidum fruiting body medium is 33.2 mg/mL.The effect of Bacillus subtilis-G.lucidum fermentation broth on the damage of intestinal mucosal barrier caused by ceftriaxone sodium was reduced,the body weight of mice recovered and colon swelling improved,colon histopathological injury was alleviated,inflammatory cell infiltration was alleviated,serum IL-10 increased significantly,LPS、TNF-αand IL-6 decreased significantly compared with model group,and the proportion of T cells and intestinal dysbiosis was improved.Conclusions The experimental results suggest that Bacillus subtilis-G.lucidum fermentation broth can effectively improve the intestinal barrier function damage,immune dysfunction and intestinal dysbiosis caused by antibiotic overdose,and has a certain regulatory effect on intestinal mucosal barrier function.展开更多
Objective:To investigate the effects of electroacupuncture on duodenal inflammation and intestinal mucosal barrier function in functional dyspepsia(FD)rats based on the Toll-like receptors 4(TLR4)/myeloid differentiat...Objective:To investigate the effects of electroacupuncture on duodenal inflammation and intestinal mucosal barrier function in functional dyspepsia(FD)rats based on the Toll-like receptors 4(TLR4)/myeloid differentiation factor 88(Myd88)pathway.Methods:Sixty male Sprague-Dawley rats were randomly divided into a blank group(n=10)and a modeling group(n=50).The FD model was induced in the modeling group using a multi-factor comprehensive intervention.Forty successful model rats were randomly allocated into a model(FD)group,an electroacupuncture(EA)group,a TLR4 inhibitor(TAK242)group,and an EA+TAK242 group,with 10 rats in each group.In the EA group,following acupuncture at Zusanli(ST36)and Taichong(LR3),a filiform needle was perpendicularly inserted 3 mm into the lateral side of Zusanli(ST36)bilaterally,positioned between the Stomach Meridian and the Gallbladder Meridian at the lateral edge of the tibia,serving as an auxiliary electrode.An EA instrument was then connected to Zusanli(ST36)and the auxiliary electrode on the same side for intervention.The TAK242 group received tail vein injections of TLR4 inhibitor TAK242[0.5 mg/(kgbw)],while the EA+TAK242 group received both TAK242 injections and EA intervention.Intragastric residual rate and small intestinal propulsion rate were observed in each group.Immunohistochemistry was used to detect tryptase expression in the duodenum;enzyme-linked immunosorbent assay was used to measure serum concentrations of 5-hydroxytryptamine(5-HT)and 5-hydroxyindole acetic acid(5-HIAA);Western blotting was used to analyze TLR4,MyD88,zonula occludens-1(ZO-1),and junctional adhesion molecule A(JAM-A)protein expression in the duodenal tissue.Results:In the FD group,the intragastric residual rate increased,small intestinal propulsion rate decreased,ZO-1 and JAM-A protein expression decreased,duodenal tryptase mean optical density increased,serum 5-HT and 5-HIAA levels increased significantly,and TLR4 and MyD88 protein expression increased significantly compared to the blank group(P<0.01).Compared to the FD group,the EA,TAK242,and EA+TAK242 groups showed a decreased gastric residual rate,increased small intestinal propulsion rate,increased ZO-1 and JAM-A protein expression,decreased optical density of duodenal tryptase,and significantly reduced serum 5-HT and 5-HIAA,as well as decreased TLR4 and MyD88 protein levels(P<0.01).Compared to the EA group,the TAK242 group had a higher intragastric residual rate and lower small intestinal propulsion rate(P<0.05),decreased TLR4 protein expression(P<0.05),no significant change in MyD88 protein expression(P>0.05),increased optical density of duodenal tryptase(P<0.05),increased serum 5-HT and 5-HIAA levels(P<0.05),and decreased ZO-1 and JAM-A protein expression(P<0.05).In the EA+TAK242 group,the intragastric residual rate was significantly lower,small intestinal propulsion rate was significantly higher,TLR4 and MyD88 protein expression was significantly lower,optical density of duodenal tryptase was significantly lower,and ZO-1 and JAM-A protein expression levels were significantly higher compared to the EA and TAK242 groups(P<0.01).Conclusion:EA promotes gastrointestinal motility,restores intestinal mucosal barrier function,and reduces inflammatory responses in FD rats,potentially through the down-regulation of the TLR4/Myd88 pathway.展开更多
基金We thank for the funding support from the National Natural Science Foundation of China(No.31900920)the Nutrition and Care of Maternal&Child Research Fund Project of Guangzhou Biostime Institute of Nutrition&Care(No.2019BINCMCF02)the Liaoning Provincial Program for Top Discipline of Basic Medical Sciences,China.
文摘Objective This study aimed to develop a type of Ganoderma lucidum(G.lucidum)-probiotic fermentation broth that can effectively improve the intestinal mucosal barrier function of mice with ceftriaxone-induced intestinal dysbiosis.Methods By means of absorbance of optical density(OD)value and phenol-concentrated sulfuric acid measurement of polysaccharide content,the probiotic species can grow on the medium of G.lucidum were screened out,and the concentration of the medium of G.lucidum was determined,and the fermentation broth was prepared for subsequent experiments.Thirty-two SPF grade male BALB/c mice were randomly divided into four groups(eight mice in each group),namely control group(CON),intestinal mucosal barrier damage model group(CS),fermentation broth intervention group(FT)and G.lucidum medium intervention group(GL),respectively.The intestinal dysregulation model was induced by intra-gastric administration of 0.2 mL ceftriaxone sodium(twice a day for seven consecutive days).From day 8,the FT group and GL group were gavage with 0.2 mL fermentation broth and G.lucidum medium,respectively.On day 15,all mice were sacrificed.To draw the weight curve and measure the cecal index;pathological examination of colon tissues with HE staining;serum levels of LPS,IL-10,TNF and IL-6 were detected by ELISA.Flow cytometry was used to analyze the changes of CD3+T cells,CD4+T cells,CD8+T cells and macrophages in spleen.16S rRNA sequencing was performed to detect the intestinal microbiota structure of mice.Results Bacillus subtilis can decompose and utilize G.lucidum fruiting body medium,and the suitable concentration of G.lucidum fruiting body medium is 33.2 mg/mL.The effect of Bacillus subtilis-G.lucidum fermentation broth on the damage of intestinal mucosal barrier caused by ceftriaxone sodium was reduced,the body weight of mice recovered and colon swelling improved,colon histopathological injury was alleviated,inflammatory cell infiltration was alleviated,serum IL-10 increased significantly,LPS、TNF-αand IL-6 decreased significantly compared with model group,and the proportion of T cells and intestinal dysbiosis was improved.Conclusions The experimental results suggest that Bacillus subtilis-G.lucidum fermentation broth can effectively improve the intestinal barrier function damage,immune dysfunction and intestinal dysbiosis caused by antibiotic overdose,and has a certain regulatory effect on intestinal mucosal barrier function.
文摘Objective:To investigate the effects of electroacupuncture on duodenal inflammation and intestinal mucosal barrier function in functional dyspepsia(FD)rats based on the Toll-like receptors 4(TLR4)/myeloid differentiation factor 88(Myd88)pathway.Methods:Sixty male Sprague-Dawley rats were randomly divided into a blank group(n=10)and a modeling group(n=50).The FD model was induced in the modeling group using a multi-factor comprehensive intervention.Forty successful model rats were randomly allocated into a model(FD)group,an electroacupuncture(EA)group,a TLR4 inhibitor(TAK242)group,and an EA+TAK242 group,with 10 rats in each group.In the EA group,following acupuncture at Zusanli(ST36)and Taichong(LR3),a filiform needle was perpendicularly inserted 3 mm into the lateral side of Zusanli(ST36)bilaterally,positioned between the Stomach Meridian and the Gallbladder Meridian at the lateral edge of the tibia,serving as an auxiliary electrode.An EA instrument was then connected to Zusanli(ST36)and the auxiliary electrode on the same side for intervention.The TAK242 group received tail vein injections of TLR4 inhibitor TAK242[0.5 mg/(kgbw)],while the EA+TAK242 group received both TAK242 injections and EA intervention.Intragastric residual rate and small intestinal propulsion rate were observed in each group.Immunohistochemistry was used to detect tryptase expression in the duodenum;enzyme-linked immunosorbent assay was used to measure serum concentrations of 5-hydroxytryptamine(5-HT)and 5-hydroxyindole acetic acid(5-HIAA);Western blotting was used to analyze TLR4,MyD88,zonula occludens-1(ZO-1),and junctional adhesion molecule A(JAM-A)protein expression in the duodenal tissue.Results:In the FD group,the intragastric residual rate increased,small intestinal propulsion rate decreased,ZO-1 and JAM-A protein expression decreased,duodenal tryptase mean optical density increased,serum 5-HT and 5-HIAA levels increased significantly,and TLR4 and MyD88 protein expression increased significantly compared to the blank group(P<0.01).Compared to the FD group,the EA,TAK242,and EA+TAK242 groups showed a decreased gastric residual rate,increased small intestinal propulsion rate,increased ZO-1 and JAM-A protein expression,decreased optical density of duodenal tryptase,and significantly reduced serum 5-HT and 5-HIAA,as well as decreased TLR4 and MyD88 protein levels(P<0.01).Compared to the EA group,the TAK242 group had a higher intragastric residual rate and lower small intestinal propulsion rate(P<0.05),decreased TLR4 protein expression(P<0.05),no significant change in MyD88 protein expression(P>0.05),increased optical density of duodenal tryptase(P<0.05),increased serum 5-HT and 5-HIAA levels(P<0.05),and decreased ZO-1 and JAM-A protein expression(P<0.05).In the EA+TAK242 group,the intragastric residual rate was significantly lower,small intestinal propulsion rate was significantly higher,TLR4 and MyD88 protein expression was significantly lower,optical density of duodenal tryptase was significantly lower,and ZO-1 and JAM-A protein expression levels were significantly higher compared to the EA and TAK242 groups(P<0.01).Conclusion:EA promotes gastrointestinal motility,restores intestinal mucosal barrier function,and reduces inflammatory responses in FD rats,potentially through the down-regulation of the TLR4/Myd88 pathway.
