数字中医药（英文）2020，Vol.3Issue(3)：199-212,14.DOI:10.1016/j.dcmed.2020.09.006
枯草芽孢杆菌-灵芝子实体发酵液改善头孢曲松钠诱导的小鼠肠道菌群失调及肠黏膜屏障功能受损
Fermentation Production of Ganoderma lucidum by Bacillus subtilis Ameliorated Ceftriaxone-induced Intestinal Dysbiosis and Improved Intestinal Mucosal Barrier Function in Mice
摘要
Abstract
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 macro-phages 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-α and 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.关键词
灵芝/枯草芽孢杆菌/传统中药发酵/灵芝酸/肠黏膜屏障功能/肠道菌群失调/T细胞调节Key words
Ganoderma lucidum/Bacillus subtilis/Traditional Chinese medicine fermentation/Ganoderma lucidum acid/Intestinal mucosal barrier function/Intestinal dysbiosis/Regulate T cell responses引用本文复制引用
刘赫,范青杰,刘银辉,李明,袁杰力..枯草芽孢杆菌-灵芝子实体发酵液改善头孢曲松钠诱导的小鼠肠道菌群失调及肠黏膜屏障功能受损[J].数字中医药（英文）,2020,3(3):199-212,14.基金项目
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) and the Liaoning Provincial Program for Top Discipline of Basic Medical Sciences, China. （No. 31900920）
