Limosilac to bacillus reuteri QS01 is potential probiotic isolated from the intestinal microbiota of healthy women in early pregnancy.In the current study,we examined whether QS01 can prevent gestational diabetes mell...Limosilac to bacillus reuteri QS01 is potential probiotic isolated from the intestinal microbiota of healthy women in early pregnancy.In the current study,we examined whether QS01 can prevent gestational diabetes mellitus(GDM) in an enhanced mouse model of the condition.Female C57BL/6J mouse offspring(F1 generation) born to dams fed a control(CON) or low-protein diet(GDM group) during gestation and lactation were used.Pregnant F1 mice fed a standard diet were randomly assigned to 5 groups:CON,GDM,and GDM mice treated with metformin,QS01 or Lacticaseibacillus rhamnosus HN001.An oral glucose tolerance test was performed before sacrifice at gestational day 17.Glucose tolerance was significantly ameliorated by all 3 treatments.QS01 supplementation fortified the intestinal mucosal barrier and inhibited the escalation of plasma inflammatory cytokines.QS01 treatment altered the cecal microbiota composition and function.Plasma and cecal metabolite profiles were modulated by QS01,prominently demonstrating significant upregulation of indole lactate and L-tryptophan in plasma and 5-hydroxy-tryptophan in the cecum,positively correlated with gut Lactobacillus abundance.In summary,QS01 plays a role in preventing GDM by remodeling the intestinal microbiota,reinforcing the intestinal mucosal barrier and alleviating chronic inflammation.展开更多
AIM To demonstrate that specific bacteria might release bacterial extracellular DNA(e DNA) to exert immunomodulatory functions in the mouse small intestine.METHODS Extracellular DNA was extracted using phosphate buffe...AIM To demonstrate that specific bacteria might release bacterial extracellular DNA(e DNA) to exert immunomodulatory functions in the mouse small intestine.METHODS Extracellular DNA was extracted using phosphate buffered saline with 0.5 mmol/L dithiothreitol combined with two phenol extractions. TOTO-1 iodide, a cell-impermeant and high-affinity nucleic acid stain, was used to confirm the existence of e DNA in the mucus layers of the small intestineand colon in healthy Male C57 BL/6 mice. Composition difference of e DNA and intracellular DNA(i DNA) of the small intestinal mucus was studied by Illumina sequencing and terminal restriction fragment length polymorphism(T-RFLP). Stimulation of cytokine production by e DNA was studied in RAW264.7 cells in vitro.RESULTS TOTO-1 iodide staining confirmed existence of e DNA in loose mucus layer of the mouse colon and thin surface mucus layer of the small intestine. Illumina sequencing analysis and T-RFLP revealed that the composition of the e DNA in the small intestinal mucus was significantly different from that of the i DNA of the small intestinal mucus bacteria. Illumina Miseq sequencing showed that the e DNA sequences came mainly from Gram-negative bacteria of Bacteroidales S24-7. By contrast, predominant bacteria of the small intestinal flora comprised Grampositive bacteria. Both e DNA and i DNA were added to native or lipopolysaccharide-stimulated Raw267.4 macrophages, respectively. The e DNA induced significantly lower tumor necrosis factor-α/interleukin-10(IL-10) and IL-6/IL-10 ratios than i DNA, suggesting the predominance for maintaining immune homeostasis of the gut.CONCLUSION Our results indicated that degraded bacterial genomic DNA was mainly released by Gram-negative bacteria, especially Bacteroidales-S24-7 and Stenotrophomonas genus in gut mucus of mice. They decreased pro-inflammatory activity compared to total gut flora genomic DNA.展开更多
基金supported by the grants from the Nutrition and care of Maternal and Child Research Funding program (2020BINCMCF056)。
文摘Limosilac to bacillus reuteri QS01 is potential probiotic isolated from the intestinal microbiota of healthy women in early pregnancy.In the current study,we examined whether QS01 can prevent gestational diabetes mellitus(GDM) in an enhanced mouse model of the condition.Female C57BL/6J mouse offspring(F1 generation) born to dams fed a control(CON) or low-protein diet(GDM group) during gestation and lactation were used.Pregnant F1 mice fed a standard diet were randomly assigned to 5 groups:CON,GDM,and GDM mice treated with metformin,QS01 or Lacticaseibacillus rhamnosus HN001.An oral glucose tolerance test was performed before sacrifice at gestational day 17.Glucose tolerance was significantly ameliorated by all 3 treatments.QS01 supplementation fortified the intestinal mucosal barrier and inhibited the escalation of plasma inflammatory cytokines.QS01 treatment altered the cecal microbiota composition and function.Plasma and cecal metabolite profiles were modulated by QS01,prominently demonstrating significant upregulation of indole lactate and L-tryptophan in plasma and 5-hydroxy-tryptophan in the cecum,positively correlated with gut Lactobacillus abundance.In summary,QS01 plays a role in preventing GDM by remodeling the intestinal microbiota,reinforcing the intestinal mucosal barrier and alleviating chronic inflammation.
基金Supported by China Postdoctoral Science Foundation,No.172774Fund of Key Laboratory of Carbohydrate Chemistry and Biotechnology,Ministry of Education,Jiangnan University,No.KLCCB-KF201603National Natural Science Foundation of China,No.31201805
文摘AIM To demonstrate that specific bacteria might release bacterial extracellular DNA(e DNA) to exert immunomodulatory functions in the mouse small intestine.METHODS Extracellular DNA was extracted using phosphate buffered saline with 0.5 mmol/L dithiothreitol combined with two phenol extractions. TOTO-1 iodide, a cell-impermeant and high-affinity nucleic acid stain, was used to confirm the existence of e DNA in the mucus layers of the small intestineand colon in healthy Male C57 BL/6 mice. Composition difference of e DNA and intracellular DNA(i DNA) of the small intestinal mucus was studied by Illumina sequencing and terminal restriction fragment length polymorphism(T-RFLP). Stimulation of cytokine production by e DNA was studied in RAW264.7 cells in vitro.RESULTS TOTO-1 iodide staining confirmed existence of e DNA in loose mucus layer of the mouse colon and thin surface mucus layer of the small intestine. Illumina sequencing analysis and T-RFLP revealed that the composition of the e DNA in the small intestinal mucus was significantly different from that of the i DNA of the small intestinal mucus bacteria. Illumina Miseq sequencing showed that the e DNA sequences came mainly from Gram-negative bacteria of Bacteroidales S24-7. By contrast, predominant bacteria of the small intestinal flora comprised Grampositive bacteria. Both e DNA and i DNA were added to native or lipopolysaccharide-stimulated Raw267.4 macrophages, respectively. The e DNA induced significantly lower tumor necrosis factor-α/interleukin-10(IL-10) and IL-6/IL-10 ratios than i DNA, suggesting the predominance for maintaining immune homeostasis of the gut.CONCLUSION Our results indicated that degraded bacterial genomic DNA was mainly released by Gram-negative bacteria, especially Bacteroidales-S24-7 and Stenotrophomonas genus in gut mucus of mice. They decreased pro-inflammatory activity compared to total gut flora genomic DNA.
