本研究通过特异性引物PCR、ERIC-PCR指纹图谱,从一健康人体肠道内筛选到20种ERIC类型的分离物。选取不同ERIC类型的代表菌株进行16S r RNA全长基因测序并在NCBI中进行BLAST,发现15种代表菌株的16S r RNA基因序列与库中的普拉梭菌（Faeca...本研究通过特异性引物PCR、ERIC-PCR指纹图谱,从一健康人体肠道内筛选到20种ERIC类型的分离物。选取不同ERIC类型的代表菌株进行16S r RNA全长基因测序并在NCBI中进行BLAST,发现15种代表菌株的16S r RNA基因序列与库中的普拉梭菌（Faecalibacterium prausnitzii）相似性均达到97%以上,初步将这些代表菌株鉴定为普拉梭菌。为了筛选出与人体健康具有密切相关性且生物学活性强的优良菌株,通过综合分析15种ERIC类型的分离物的分离丰度及遗传距离,从鉴定出的15株代表菌株中挑选了7株候选优良菌株,进行胆汁酸盐耐受、丁酸盐产生和对肠屏障功能的障影响等评价。研究表明：胆汁酸盐浓度高于0.1%时,F18、F22、F109和F139菌株仍然可以生长,表现出较强的耐胆汁酸能力;菌株间产丁酸盐能力差异显著,F31的丁酸盐产量最高,其次为F18、F20、F33和F139,且这4株菌株间产丁酸盐能力无显著差异;菌株F31使肠屏障功能受损,而其他菌株无明显影响。综合实验数据,筛选出F18和F139为优良菌株。本研究成功实现了普拉梭菌的分离鉴定及优良菌株筛选,为后续的体内实验研究提供了重要研究基础。展开更多
AIM: To explore the preventive and therapeutic effects of Faecalibacterium prausnitzii(F. prausnitzii) supernatant on dextran sulfate sodium(DSS) induced colitis in mice.METHODS: Forty C57BL/6J male mice were randomly...AIM: To explore the preventive and therapeutic effects of Faecalibacterium prausnitzii(F. prausnitzii) supernatant on dextran sulfate sodium(DSS) induced colitis in mice.METHODS: Forty C57BL/6J male mice were randomlydivided into four groups: control group, model group, treatment group, and prevention group. Mice were weighed daily. On day 10, the colon length was measured, the colorectal histopathologic damage score(HDS) was assessed, and plasma interleukin(IL)-17 A, IL-6, and IL-4 levels were detected by enzyme-linked immunosorbent assay. The expression of transcription factor retinoic acid-related orphan receptor-γt(RORγt) and IL-17 A in colon inflammatory mucosa tissue were determined by immunohistochemical assay, and the expression levels of RORγt m RNA, IL-17 A m RNA, and IL-6 m RNA were detected by real-time quantitative polymerase chain reaction(PCR). The proportion of Th17 in mononuclear cells in spleen was assayed by fluorescence activated cell sorter. RESULTS: When compared with the model group, the colon length(P < 0.05) and body weight(P < 0.01) in the treatment and prevention groups were significantly increased, and the colon HDS was decreased(P < 0.05 and P < 0.01). There was no statistical difference between the treatment group and prevention group. After treatment with F. prausnitzii supernatant, the plasma levels of IL-17 A and IL-6(P < 0.05), the protein and m RNA expression of IL-17 A and RORγt, and the Th17 cell ratio of spleen cells(P < 0.01) were significantly decreased compared to the model group. Plasma IL-4 level in the prevention group was significantly higher than that in the model group(P < 0.05), but there was no significant difference between these two groups in the expression of IL-6 in both the plasma and colon mucosa tissues.CONCLUSION: F. prausnitzii supernatant exerts protective and therapeutic effects on DSS-induced colitis in mice, probably via inhibition of Th17 differentiation and IL- 17A secretion in the plasma and colon mucosa tissues. It can also improve colitis in mice by downregulating IL-6 and prevent colitis by upregulating IL-4.展开更多
Objective: To study the effects of Faecalibacterium prausnitzii intervention on immune response, intestinal flora and intestinal mucosal barrier of mice with ulcerative colitis (UC). Methods: C57BL/6J mice were random...Objective: To study the effects of Faecalibacterium prausnitzii intervention on immune response, intestinal flora and intestinal mucosal barrier of mice with ulcerative colitis (UC). Methods: C57BL/6J mice were randomly divided into control group, UC group and Faecalibacterium prausnitzii group, the latter two groups were made into UC models by trinitrobenzene sulfonic acid enema and F. prausnitzii group were given intragastric administration of F. prausnitzii solution for intervention. The differences in immune response, intestinal flora, and intestinal mucosal barrier were compared among the three groups after 7 days of intervention. Results: The interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) contents in serum, the fork head box P3 (Foxp3), zonula occludens-1 (ZO-1), occludin, claudin-1 and claudin-2 expression in intestinal mucosa as well as the number of bifidobacterium and lactobacillus in feces of the UC group were significantly lower than those of the control group whereas the interleukin-17 (IL-17), diamine oxidase (DAO) and D-lactic acid (D-LA) contents in serum, the retinoid-related orphan nuclear receptor γt (RORγt) expression in intestinal mucosa as well as the number of enterobacter and enterococcus in feces were significantly higher than those of the control group (P<0.05);IL-10 and TGF-β1 contents in serum, Foxp3, ZO-1, occludin, claudin-1 and claudin-2 expression in intestinal mucosa as well as the number of bifidobacterium and lactobacillus in feces of the F. prausnitzii group were significantly higher than those of the UC group whereas IL-17, DAO and D-LA contents in serum, RORγt expression in intestinal mucosa as well as the number of enterobacter and enterococcus in feces were significantly lower than those of the UC group (P<0.05). Conclusion: Faecalibacterium prausnitzii intervention can improve the Th17/Treg immune response, intestinal flora and intestinal mucosal barrier in UC mice.展开更多
Sleep deprivation(SD)is a widespread public health concern associated with cognitive dysfunction and neu-roinflammation,but effective interventions are still limited.In sleep-deprived mice,a notable reduction in Faeca...Sleep deprivation(SD)is a widespread public health concern associated with cognitive dysfunction and neu-roinflammation,but effective interventions are still limited.In sleep-deprived mice,a notable reduction in Faecalibacterium was observed.However,the potential of this probiotic to treat cognitive impairment caused by SD has not yet been investigated.In this study,we investigated whether supplementation with Faecalibacterium prausnitzii could alleviate cognitive dysfunction induced by SD.We demonstrated that F.prausnitzii colonization ameliorated SD-induced cognitive impairment and hippocampal neuron damage in a 72-h SD mouse model.Metabolomics analysis revealed that differential metabolites in SD mice supplemented with F.prausnitzii were significantly enriched in pathways including the synaptic vesicle cycle and neuroactive ligand-receptor inter-action,with a particularly notable decrease inγ-aminobutyric acid(GABA)levels.Transcriptomic profiling identified GABAergic and cholinergic synaptic signaling pathways as the most significantly enriched pathways differentiating SD mice with and without probiotic intervention.Integrated analysis revealed significant corre-lations between metabolites,including GABA,and the transcriptional profiles of genes within the GABAergic and cholinergic signaling pathways.Furthermore,pre-colonization with F.prausnitzii was found to prevent SD-induced hippocampal neuronal loss,increase the levels of key synaptic proteins(Psd-95,Syp,and Snap-25),attenuate microglial overactivation,and suppress pro-inflammatory factor expression.These results indicate that F.prausnitzii supplementation mitigates cognitive impairment,synaptic damage,and neuroinflammation caused by SD.Overall,our work provides novel insights into the role of a specific gut microbe in SD-induced neural impairment by systematically elucidating the GABA-mediated gut-brain axis mechanism through an in-tegrated multi-omics approach.展开更多
Revealing strain-level diversity in the gut microbiome is critical to understanding its impact on health.Genomic and metabolomic advances enable strain-level dissection of the gut microbiome.Faecalibacterium prausnitz...Revealing strain-level diversity in the gut microbiome is critical to understanding its impact on health.Genomic and metabolomic advances enable strain-level dissection of the gut microbiome.Faecalibacterium prausnitzii,a prevalent commensal bacterium in the human gut,is a promising candidate for next-generation probiotics.However,the study on genomic and metabolomic diversity of F.prausnitzii strains remains insufficient.In this study,12 F.prausnitzii strains were isolated and whole genome sequencing was conducted.Results showed that these strains were classified into three phylogenetic clades(clade B,C,and D).Additionally,strain FP3003 displayed typical characteristics,marked by significant acetate consumption and butyrate production.Further integration of non-targeted metabolomics analysis and an in vitro intestinal epithelial cell model confirmed the established intestinal protective effects of F.prausnitzii and butyrate.Moreover,we identified four novel beneficial F.prausnitzii-derived metabolites,including indolelactic acid(ILA),hydroxyphenyllactic acid,hydroxyisocaproic acid,and butoxyacetic acid,among which ILA upregulated Occludin and Ecadherin expression in vitro.In summary,our results systematically elucidated the strain-level insights into the metabolic diversity and functional heterogeneity of F.prausnitzii.This study expanded the current knowledge on F.prausnitzii and established a key research paradigm for investigating the functional potential of next-generation probiotics.展开更多
Hypertriglyceridemia(HTG)is a common and significant contributor to acute pancreatitis(AP).Caffeine has been reported to have a protective effect against pancreatic disorders.However,the role of caffeine in hypertrigl...Hypertriglyceridemia(HTG)is a common and significant contributor to acute pancreatitis(AP).Caffeine has been reported to have a protective effect against pancreatic disorders.However,the role of caffeine in hypertriglyceridemia acute pancreatitis(HAP)is still unknown.To investigate a new understanding of the relationship between caffeine and gut microbiota in HAP development.Using multi-omics analysis of 71 HAP and HTG patients,their characteristics of intestinal microecology were detected.HTG mice models were established through either pharmacological induction(P-407 intraperitoneal injection)or genetic ablation of GPIHBP1(GPIHBP1^(−/−)).Genes deferentially expressed in the gut were identified by RNA sequencing and the role of caffeine in reshaping gut microbial networks was investigated.Caffeine levels decreased significantly in HAP patients and were inversely correlated with the severity of HAP.Caffeine maintained the intestinal homeostasis and attenuated HAP through a gut microbiota-dependent manner in the mouse model.Specially,the commensal Faecalibacterium prausnitzii,which contributed most in distinguishing the differences between HTG and HAP patients,has been found to be related to the effect of caffeine on alleviating HAP.RNA sequencing combined with TLR4^(−/−)mice revealed that TLR4/NLRP3 may be the key signaling pathway for caffeine to maintain intestinal homeostasis and alleviating HAP.This study reveals the gut metabolites and microbiota characteristics of HAP and HTG patients,and explores the therapeutic potential of caffeine against HAP from the perspective of gut microbiota.展开更多
Background:Accumulating evidence suggests that metabolic disorders,including type 2 diabetes mellitus(T2DM),can be treated with traditional Chinese medicine formulas,such as the Gegen Qinlian decoction(GQD).This study...Background:Accumulating evidence suggests that metabolic disorders,including type 2 diabetes mellitus(T2DM),can be treated with traditional Chinese medicine formulas,such as the Gegen Qinlian decoction(GQD).This study elucidates the mechanisms by which gut microbes mediate the anti-diabetic effects of GQD.Methods:We conducted a double-blind randomized clinical trial involving 120 untreated participants with T2DM.During the 12-week intervention,anthropometric measurements and diabetic traits were recorded every 4 weeks.Fecal microbiota and serum metabolites were measured before and after the intervention using 16S rDNA sequencing,liquid chromatography-mass spectrometry,and Bio-Plex panels.Results:Anti-diabetic effects were observed in the GQD group in the human trial.Specifically,glycated hemoglobin,fasting plasma glucose,and two-hour postprandial blood glucose levels were significantly lower in the GQD group than in the placebo group.Additionally,Faecalibacterium was significantly enriched in the GQD group,and the short-chain fatty acid levels were higher and the serum inflammation-associated marker levels were lower in the GQD group compared to the placebo group.Moreover,Faecalibacterium abundance negatively correlated with the levels of serum hemoglobin,fasting plasma glucose,and pro-inflammatory cytokines.Finally,the diabetes-alleviating effect of Faecalibacterium was confirmed by oral administration of Faecalibacterium prausnitzi(DSMZ 17677)in T2DMmousemodel.Conclusions:GQD improved type 2 diabetes primarily by modulating the abundance of Faecalibacterium in the gut microbiota,alleviating metabolic disorders and the inflammatory state.展开更多
文摘本研究通过特异性引物PCR、ERIC-PCR指纹图谱,从一健康人体肠道内筛选到20种ERIC类型的分离物。选取不同ERIC类型的代表菌株进行16S r RNA全长基因测序并在NCBI中进行BLAST,发现15种代表菌株的16S r RNA基因序列与库中的普拉梭菌（Faecalibacterium prausnitzii）相似性均达到97%以上,初步将这些代表菌株鉴定为普拉梭菌。为了筛选出与人体健康具有密切相关性且生物学活性强的优良菌株,通过综合分析15种ERIC类型的分离物的分离丰度及遗传距离,从鉴定出的15株代表菌株中挑选了7株候选优良菌株,进行胆汁酸盐耐受、丁酸盐产生和对肠屏障功能的障影响等评价。研究表明：胆汁酸盐浓度高于0.1%时,F18、F22、F109和F139菌株仍然可以生长,表现出较强的耐胆汁酸能力;菌株间产丁酸盐能力差异显著,F31的丁酸盐产量最高,其次为F18、F20、F33和F139,且这4株菌株间产丁酸盐能力无显著差异;菌株F31使肠屏障功能受损,而其他菌株无明显影响。综合实验数据,筛选出F18和F139为优良菌株。本研究成功实现了普拉梭菌的分离鉴定及优良菌株筛选,为后续的体内实验研究提供了重要研究基础。
基金Supported by National Natural Science Foundation of China,No.81470819
文摘AIM: To explore the preventive and therapeutic effects of Faecalibacterium prausnitzii(F. prausnitzii) supernatant on dextran sulfate sodium(DSS) induced colitis in mice.METHODS: Forty C57BL/6J male mice were randomlydivided into four groups: control group, model group, treatment group, and prevention group. Mice were weighed daily. On day 10, the colon length was measured, the colorectal histopathologic damage score(HDS) was assessed, and plasma interleukin(IL)-17 A, IL-6, and IL-4 levels were detected by enzyme-linked immunosorbent assay. The expression of transcription factor retinoic acid-related orphan receptor-γt(RORγt) and IL-17 A in colon inflammatory mucosa tissue were determined by immunohistochemical assay, and the expression levels of RORγt m RNA, IL-17 A m RNA, and IL-6 m RNA were detected by real-time quantitative polymerase chain reaction(PCR). The proportion of Th17 in mononuclear cells in spleen was assayed by fluorescence activated cell sorter. RESULTS: When compared with the model group, the colon length(P < 0.05) and body weight(P < 0.01) in the treatment and prevention groups were significantly increased, and the colon HDS was decreased(P < 0.05 and P < 0.01). There was no statistical difference between the treatment group and prevention group. After treatment with F. prausnitzii supernatant, the plasma levels of IL-17 A and IL-6(P < 0.05), the protein and m RNA expression of IL-17 A and RORγt, and the Th17 cell ratio of spleen cells(P < 0.01) were significantly decreased compared to the model group. Plasma IL-4 level in the prevention group was significantly higher than that in the model group(P < 0.05), but there was no significant difference between these two groups in the expression of IL-6 in both the plasma and colon mucosa tissues.CONCLUSION: F. prausnitzii supernatant exerts protective and therapeutic effects on DSS-induced colitis in mice, probably via inhibition of Th17 differentiation and IL- 17A secretion in the plasma and colon mucosa tissues. It can also improve colitis in mice by downregulating IL-6 and prevent colitis by upregulating IL-4.
基金Natural Science Basic Research Program of Shaanxi (2017JM8113)
文摘Objective: To study the effects of Faecalibacterium prausnitzii intervention on immune response, intestinal flora and intestinal mucosal barrier of mice with ulcerative colitis (UC). Methods: C57BL/6J mice were randomly divided into control group, UC group and Faecalibacterium prausnitzii group, the latter two groups were made into UC models by trinitrobenzene sulfonic acid enema and F. prausnitzii group were given intragastric administration of F. prausnitzii solution for intervention. The differences in immune response, intestinal flora, and intestinal mucosal barrier were compared among the three groups after 7 days of intervention. Results: The interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) contents in serum, the fork head box P3 (Foxp3), zonula occludens-1 (ZO-1), occludin, claudin-1 and claudin-2 expression in intestinal mucosa as well as the number of bifidobacterium and lactobacillus in feces of the UC group were significantly lower than those of the control group whereas the interleukin-17 (IL-17), diamine oxidase (DAO) and D-lactic acid (D-LA) contents in serum, the retinoid-related orphan nuclear receptor γt (RORγt) expression in intestinal mucosa as well as the number of enterobacter and enterococcus in feces were significantly higher than those of the control group (P<0.05);IL-10 and TGF-β1 contents in serum, Foxp3, ZO-1, occludin, claudin-1 and claudin-2 expression in intestinal mucosa as well as the number of bifidobacterium and lactobacillus in feces of the F. prausnitzii group were significantly higher than those of the UC group whereas IL-17, DAO and D-LA contents in serum, RORγt expression in intestinal mucosa as well as the number of enterobacter and enterococcus in feces were significantly lower than those of the UC group (P<0.05). Conclusion: Faecalibacterium prausnitzii intervention can improve the Th17/Treg immune response, intestinal flora and intestinal mucosal barrier in UC mice.
基金funded by the 111 project of the Education Ministry of China(No.B18053)China Postdoctoral Science Foundation(2023M7337942024T171009).
文摘Sleep deprivation(SD)is a widespread public health concern associated with cognitive dysfunction and neu-roinflammation,but effective interventions are still limited.In sleep-deprived mice,a notable reduction in Faecalibacterium was observed.However,the potential of this probiotic to treat cognitive impairment caused by SD has not yet been investigated.In this study,we investigated whether supplementation with Faecalibacterium prausnitzii could alleviate cognitive dysfunction induced by SD.We demonstrated that F.prausnitzii colonization ameliorated SD-induced cognitive impairment and hippocampal neuron damage in a 72-h SD mouse model.Metabolomics analysis revealed that differential metabolites in SD mice supplemented with F.prausnitzii were significantly enriched in pathways including the synaptic vesicle cycle and neuroactive ligand-receptor inter-action,with a particularly notable decrease inγ-aminobutyric acid(GABA)levels.Transcriptomic profiling identified GABAergic and cholinergic synaptic signaling pathways as the most significantly enriched pathways differentiating SD mice with and without probiotic intervention.Integrated analysis revealed significant corre-lations between metabolites,including GABA,and the transcriptional profiles of genes within the GABAergic and cholinergic signaling pathways.Furthermore,pre-colonization with F.prausnitzii was found to prevent SD-induced hippocampal neuronal loss,increase the levels of key synaptic proteins(Psd-95,Syp,and Snap-25),attenuate microglial overactivation,and suppress pro-inflammatory factor expression.These results indicate that F.prausnitzii supplementation mitigates cognitive impairment,synaptic damage,and neuroinflammation caused by SD.Overall,our work provides novel insights into the role of a specific gut microbe in SD-induced neural impairment by systematically elucidating the GABA-mediated gut-brain axis mechanism through an in-tegrated multi-omics approach.
基金supported by the National Natural Science Foundation of China for the Key Project of International Cooperative Research(32120103012)the Young Scientists Fund of the National Natural Science Foundation of China(32402085)+1 种基金the Technological Project of Jiangxi Province(20232BCD44003)the Postdoctoral Fellowship Program and China Postdoctoral Science Foundation(2024M761261).
文摘Revealing strain-level diversity in the gut microbiome is critical to understanding its impact on health.Genomic and metabolomic advances enable strain-level dissection of the gut microbiome.Faecalibacterium prausnitzii,a prevalent commensal bacterium in the human gut,is a promising candidate for next-generation probiotics.However,the study on genomic and metabolomic diversity of F.prausnitzii strains remains insufficient.In this study,12 F.prausnitzii strains were isolated and whole genome sequencing was conducted.Results showed that these strains were classified into three phylogenetic clades(clade B,C,and D).Additionally,strain FP3003 displayed typical characteristics,marked by significant acetate consumption and butyrate production.Further integration of non-targeted metabolomics analysis and an in vitro intestinal epithelial cell model confirmed the established intestinal protective effects of F.prausnitzii and butyrate.Moreover,we identified four novel beneficial F.prausnitzii-derived metabolites,including indolelactic acid(ILA),hydroxyphenyllactic acid,hydroxyisocaproic acid,and butoxyacetic acid,among which ILA upregulated Occludin and Ecadherin expression in vitro.In summary,our results systematically elucidated the strain-level insights into the metabolic diversity and functional heterogeneity of F.prausnitzii.This study expanded the current knowledge on F.prausnitzii and established a key research paradigm for investigating the functional potential of next-generation probiotics.
基金the National Natural Science Foundation of China(82470675,82270671,81970555,82400752,82300731,82200714)the Xinyi Digestive Disease Research Fund(KY-2023-03-01)the Songjiang District Science and Technology Research Project(22SJKJGG27).
文摘Hypertriglyceridemia(HTG)is a common and significant contributor to acute pancreatitis(AP).Caffeine has been reported to have a protective effect against pancreatic disorders.However,the role of caffeine in hypertriglyceridemia acute pancreatitis(HAP)is still unknown.To investigate a new understanding of the relationship between caffeine and gut microbiota in HAP development.Using multi-omics analysis of 71 HAP and HTG patients,their characteristics of intestinal microecology were detected.HTG mice models were established through either pharmacological induction(P-407 intraperitoneal injection)or genetic ablation of GPIHBP1(GPIHBP1^(−/−)).Genes deferentially expressed in the gut were identified by RNA sequencing and the role of caffeine in reshaping gut microbial networks was investigated.Caffeine levels decreased significantly in HAP patients and were inversely correlated with the severity of HAP.Caffeine maintained the intestinal homeostasis and attenuated HAP through a gut microbiota-dependent manner in the mouse model.Specially,the commensal Faecalibacterium prausnitzii,which contributed most in distinguishing the differences between HTG and HAP patients,has been found to be related to the effect of caffeine on alleviating HAP.RNA sequencing combined with TLR4^(−/−)mice revealed that TLR4/NLRP3 may be the key signaling pathway for caffeine to maintain intestinal homeostasis and alleviating HAP.This study reveals the gut metabolites and microbiota characteristics of HAP and HTG patients,and explores the therapeutic potential of caffeine against HAP from the perspective of gut microbiota.
基金supported by the National Natural Science Foundation of China (Grants No.81430097,81973837,82004242,82274343 and 31771481)the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine (Grant No.ZYYCXTD-D-202001).
文摘Background:Accumulating evidence suggests that metabolic disorders,including type 2 diabetes mellitus(T2DM),can be treated with traditional Chinese medicine formulas,such as the Gegen Qinlian decoction(GQD).This study elucidates the mechanisms by which gut microbes mediate the anti-diabetic effects of GQD.Methods:We conducted a double-blind randomized clinical trial involving 120 untreated participants with T2DM.During the 12-week intervention,anthropometric measurements and diabetic traits were recorded every 4 weeks.Fecal microbiota and serum metabolites were measured before and after the intervention using 16S rDNA sequencing,liquid chromatography-mass spectrometry,and Bio-Plex panels.Results:Anti-diabetic effects were observed in the GQD group in the human trial.Specifically,glycated hemoglobin,fasting plasma glucose,and two-hour postprandial blood glucose levels were significantly lower in the GQD group than in the placebo group.Additionally,Faecalibacterium was significantly enriched in the GQD group,and the short-chain fatty acid levels were higher and the serum inflammation-associated marker levels were lower in the GQD group compared to the placebo group.Moreover,Faecalibacterium abundance negatively correlated with the levels of serum hemoglobin,fasting plasma glucose,and pro-inflammatory cytokines.Finally,the diabetes-alleviating effect of Faecalibacterium was confirmed by oral administration of Faecalibacterium prausnitzi(DSMZ 17677)in T2DMmousemodel.Conclusions:GQD improved type 2 diabetes primarily by modulating the abundance of Faecalibacterium in the gut microbiota,alleviating metabolic disorders and the inflammatory state.
