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Purpose: When examining the underlying processes of obesity, evaluation of gut flora and energy homeostasis can be crucial since disruption of the normal gut microbiota community and energy imbalances are significant factors in the development of obesity. Therefore, this study aimed to compare the relative abundance of important obesity modulator gut microbiota (including Firmicutes, Bacteroidetes, Bifidobactrium spp., Lactobacillus spp., Bacteroides fragilis, Faecalibacterium prausnitzii, Akkermansia muciniphila, and Escherichia coli) in fecal samples of normometabilic and hypometabolic overweight/obese individuals. Methods: This matched case-control study conducted on 36 healthy women aged 18-50 years old. An indirect calorimeter and impedance body analyzer were used to assess resting metabolic rate (RMR) and body composition, respectively. Dietary intake and physical activity were assessed using questionnaires. To determine the abundance of the abovementioned gut microbiota, quantitative polymerase chain reaction (qPCR) method was performed. Moreover, ELISA kits were used to assess leptin, ghrelin, and insulin hormones. Results: The results highlighted higher load of Firmicutes (p = 0.02), F. prausnitzii (p < 0.001), and B. fragilis (p = 0.02) in the normometabolic individuals compared to the hypometabolic ones. Besides, the positive correlation between the abundance of Firmicutes (ß = 7.76 × 10-1, p = 0.01), F. prausnitzii (ß = 1.29 × 10-5, p = 0.01), and B. fragilis (ß = 4.13 × 10-6, p = 0.04) with the RMR have been shown. Whereas the abundance of Bacteroidetes, A. muciniphila, Lactobacillus spp., Bifidobactrium spp., and E. coli showed no significant difference (p > 0.05) and no significant correlation with the RMR except Lactobacillus spp. (ß = 1.73 × 10-4, p = 0.01). Conclusion: It seems that gut microbiota can be a potential target for refining host energy homeostasis and treating obesity and its consequences.
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PURPOSE: The intestine has substantial role in cholesterol homeostasis due to the presence of various cholesterol transporters and gut microbiota. Bacteroides spp. are important members of gut microbiota that employ outer membrane vesicles (OMVs) to interact with host. In this regard, we evaluated the effect of Bacteroides fragilis, Bacteroides thetaiotaomicron and related OMVs on the gene expression of important cholesterol transporters, Niemann-Pick C1-Like 1 (NPC1L1), ATP-binding cassette (ABCA1), and liver X receptors (LXRs) in Caco-2 cells. METHODS: OMVs were isolated from overnight brain heart infusion (BHI) broth of bacterial standard strains using deoxycholate and assessed by Scanning electron microscopy (SEM). The relative change in genes expression was assessed by Quantitative reverse transcription PCR (RT-qPCR) based on SYBR Green and 2-∆∆ct method in Caco-2 cells that were treated with bacteria and OMVs. Data were statistically analyzed with GraphPad Prism software. Finally, pathway enrichment based on the studied genes was performed using Cytoscape plugin ClueGO. RESULTS: B. fragilis (P value = 0.002) and B. thetaiotaomicron (P value = 0.001) significantly reduced NPC1L1 gene expression in Caco-2 cells. Interestingly, NPC1L1 transcripts were significantly increased by both OMVs(P value = 0.04) (P value = 0.01). Also, LXRß was significantly down regulated by B. thetaiotaomicron (P value = 0.02). ClueGO analysis on the studied genes demonstrated several functional groups which involve in lipid and cholesterol metabolism. CONCLUSION: The opposite effect of B. fragilis, B. thetaiotaomicron and related OMVs on the NPC1L1 gene expression was observed in Caco-2 cells. Interestingly, these effects partially were in line with the alternation of LXRs expression. However, based on pathway enrichment analysis, further molecular investigations are required to elaborate in details the specific association between Bacteroides spp. and OMVs with regulation of cholesterol signaling pathways including cholesterol transport, lipid storage, lipid homeostasis and cholesterol homeostasis.
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AIM: The aim of present study is to investigate the effect of fatty acids on the outer membrane vesicles (OMVs) produced by Bacteroides spp. BACKGROUND: Bacteroides spp. is the important member of Gut microbiota that employ OMVs production for interact with host. Besides, dietary fatty acids could influence on determination of gut microbiota composition and immune response. In this regard, we evaluated the effect of fatty acids on the growth and OMVs production of Bacteroides fragilis and Bacteroides thetaiotaomicron. METHODS: B. fragilis and B. thetaiotaomicron were grown on BHI broth with and without palmitic and palmitoleic acids as saturated and unsaturated fatty acids, respectively. OMVs were extracted using multiple centrifugation and tris-ethylene diamine tetra acetic acid (EDTA)-Sodium deoxy cholate buffers. Physicochemical properties of OMVs were detected by electron microscopy (SEM), Bradford Coomassie brilliant blue assay and SDS-PAGE. Data were analyzed with One-way ANOVA using SPSS. RESULTS: The growths of both Bacteroides were significantly increased by palmitic acid. Nevertheless, palmitoleic acid had no significant effect on them. Palmitic acid significantly decreased and increased the production of B. fragilis OMVs at low and high concentration, respectively. However, the production of B. thetaiotaomicron OMVs was not significantly affected by palmitic acid. Although palmitoleic acid had a significant decreasing effect on the production of B. fragilis OMVs, it significantly increased the production of B. thetaiotaomicron OMVs at low concentration. CONCLUSION: In conclusion we reported that palmitic acid had a stimulatory effect on the growth of B. fragilis and B. thetaiotaomicron and had a dose dependent effect on the production of B. fragilis OMVs. Also producing of B. thetaiotaomicron OMVs was affected by palmitoleic acid in a dose dependent manner.
