Intermittent Fasting Promotes White Adipose Browning and Decreases Obesity by Shaping the Gut Microbiota

“Assigned reads were piped into Bowtie2 (Langmead and Salzberg, 2012) which provided functional information to the taxonomic assigned reads with the ChocoPhlAn database.”

“LEfSe (Segata et al., 2011) was used to obtain statistically significant and biologically relevant pathways from pathway abundance file.”

“Unassigned reads were piped into DIAMOND which converts the DNA reads into protein sequences and aligns them to a protein database to obtain functional information (Buchfink et al., 2015).”

“PICRUSt analysis was done on the .biom file resulting in a pathway abundance file (Langille et al., 2013).”

“16S rRNA gene amplicon sequence results were analyzed using the MOTHUR platform (Kozich et al., 2013) and aligned to SILVA and Green Genes databases separately.”

“In addition, both BAT and WAT express protonlinked monocarboxylate transporter 1 (MCT1) encoded by Mct1, which drives acetate and lactate transport across the plasma membrane of adipocytes (den Besten et al., 2013; Iwanaga et al., 2009).”

“Gut Microbiota Orchestrates EODF-Induced WAT Beiging Recently, correlative evidence revealed a metabolic interaction between the gut microbial communities and the host, and gut bacteria have an important role in the regulation of brown and beige adipose tissues (Chevalier et al., 2015; Holmes et al., 2012; Mestdagh et…”

“Gut Microbiota Orchestrates EODF-Induced WAT Beiging Recently, correlative evidence revealed a metabolic interaction between the gut microbial communities and the host, and gut bacteria have an important role in the regulation of brown and beige adipose tissues (Chevalier et al., 2015; Holmes et al., 2012; Mestdagh et…”

“…or EODF/MT (EODF versus AL or EODF/MT versus AL/MT drunk with the same water), and red letters show the effects of microbiota depletion (AB versus CV within the same feeding regimen). promote thermogenesis and suppresses obesity (Ishibashi and Seale, 2010; Shabalina et al., 2013; Wu et al., 2012).”

“Given that activation of beige cells can suppress obesity and metabolic disease (Harms and Seale, 2013), the effect of an EODF regimen on diet-induced obesity (DIO) mice was explored.”

“…relatedmetabolic disorders are growing health challenges for Western countries; they mainly result from an imbalance between energy intake and energy expenditure (Hill et al., 672 Cell Metabolism 26, 672–685, October 3, 2017 Published by Else 2012; Hossain et al., 2007; Spiegelman and Flier, 2001).”

“This pipeline uses the MetaPlAn 2 (Truong et al., 2015) software to assign taxonomic information to the Hiseq reads being sequenced.”

“…Models: Organisms/Strains Mouse: C57BL/6 Charles River Laboratories Strain Code: 027 Mouse: Ppara wild-type (Ppara+/+): C57BL/6 (Lee et al., 1995) N/A Mouse: Ppara-null (Ppara / ): C57BL/6 (Lee et al., 1995) N/A Oligonucleotides Primers for qPCR, see Table S1 This paper…”

“, 2012; Harms and Seale, 2013) and the adaptive fasting response (Inagaki et al., 2007).”

“Previous studies revealed that fasting and feeding rhythms significantly alter the gut microbiota (Secor and Carey, 2016; Thaiss et al., 2014) and that major changes in microbiota composition can directly promote WAT beiging (Chevalier et al., 2015).”

“Recently, intermittent fasting was demonstrated to optimize energy metabolism and promote health (Fontana and Partridge, 2015; Longo and Mattson, 2014).”

“Recently, intermittent fasting was demonstrated to optimize energy metabolism and promote health (Fontana and Partridge, 2015; Longo and Mattson, 2014).”

“A phylogenic tree and Operation Taxonomic Unit (OTU) table were obtained from the Mothur Bayesian classifier (Kozich et al., 2013) and uploaded to R studio for GUnifrac analysis (Chen et al., 2012).”

“The mechanism by which EODF induces higher energy expenditure might differ from that of cold exposure; cold exposure is closely tied to b-AR signaling (Cypess et al., 2015; Mund and Frishman, 2013), while fasting is known to suppress norepinephrine turnover (Knehans and Romsos, 1983) and norepinephrine-induced thermog…”

“Non-shivering thermogenesis is mainly mediated by activating brown and/or beige adipocytes (Harms and Seale, 2013; Kajimura et al., 2015).”

“Microbiota transplantation were done according to a previous study (Chevalier et al., 2015).”

“Emerging evidence suggests that non-shivering thermogenesis can re-establish energy balance and therefore counter the effects of elevated energy intake (Cannon and Nedergaard, 2011).”

“…or EODF/MT (EODF versus AL or EODF/MT versus AL/MT drunk with the same water), and red letters show the effects of microbiota depletion (AB versus CV within the same feeding regimen). promote thermogenesis and suppresses obesity (Ishibashi and Seale, 2010; Shabalina et al., 2013; Wu et al., 2012).”

“Gut Microbiota Contribute to the Beneficial Effects of EODF on DIO Mice Gut microbiota play a critical role in energy metabolism and lipid homeostasis, and germfree or microbiota-depleted rodents have decreased susceptibility to diet-induced obesity andmetabolic syndrome (B€ ackhed et al., 2007; Rabot et al., 2010; Su…”

“In addition, PPARa is known to mediate acute fasting-induced upregulation of both hepatic Fgf21 mRNA and circulating FGF21 protein levels (Montagner et al., 2016).”

“Although many agents and treatments were reported to induce browning (Bonet et al., 2013; Jiang et al., 2015; Wu et al., 2013), it is unclear how these approaches can translate to clinically relevant therapies for human metabolic disease (Nedergaard and Cannon, 2014).”

“EODF-Induced WAT Beiging is Independent of FGF21 Signaling In addition to b-AR signaling, FGF21 is another classical signaling hormone known to regulate WAT browning (Dutchak et al., 2012; Harms and Seale, 2013) and the adaptive fasting response (Inagaki et al.”

“Given current thinking that adult humans do not have active BAT (Kajimura et al., 2015; Nedergaard and Cannon, 2014), conversion of white fat to beige fat rather than BAT activation would hold more therapeutic potential.”

“Gut Microbiota Orchestrates EODF-Induced WAT Beiging Recently, correlative evidence revealed a metabolic interaction between the gut microbial communities and the host, and gut bacteria have an important role in the regulation of brown and beige adipose tissues (Chevalier et al., 2015; Holmes et al., 2012; Mestdagh et…”

“Brown adipocyte activation by cold exposure or by b-AR agonists is clinically non-feasible, and adult human BAT is largely composed of beige-like adipocytes (Shinoda et al., 2015).”

“Suárez-Zamorano et al., 2015).”

“In contrast, intermittent fasting (including EODF) has been practiced in humans, especially in certain religious groups (Patterson et al., 2015), for centuries and has been proven to benefit body composition and health (Eshghinia and Mohammadzadeh, 2013; López-Bueno et al., 2014; Patterson et al.,…”

“Cold acclimation was originally found to increase the number of brown adipocytes in the parametrical fat pad in mice (Young et al., 1984).”

“While many browning strategies have been described, mainly in rodent models, only a very limited number of them have so far supported selective browning of WAT (Bonet et al., 2013; Wu et al., 2013).”

“Intriguingly, recent studies indicate that both acetate and lactate are beiging inducers (Hanatani et al., 2016; Kim et al., 2017; Sahuri-Arisoylu et al., 2016).”

“In addition, PPARa is known to mediate acute fasting-induced upregulation of both hepatic Fgf21 mRNA and circulating FGF21 protein levels (Montagner et al., 2016).”

“Indirect calorimetry was performed on mice after 14 cycles of EODF treatment using a 12-chamber Environment Controlled CLAMS (Columbus Instruments, Columbus, OH) with one mouse/chamber as previously described (Abreu-Vieira et al., 2015).”

“Given recent evidence that the expression of Mct1 is controlled by physiological stimuli of beiging (Carrière et al., 2014; De Matteis et al., 2013), the selective upregulation of Mct1 was noted in inguinal WAT, but not in the BAT of either long-term or short-term EODF-treated mice (Figures 4E–4G).”

“Previous studies revealed that fasting and feeding rhythms significantly alter the gut microbiota (Secor and Carey, 2016; Thaiss et al., 2014) and that major changes in microbiota composition can directly promote WAT beiging (Chevalier et al., 2015).”

“, 2015), for centuries and has been proven to benefit body composition and health (Eshghinia and Mohammadzadeh, 2013; López-Bueno et al., 2014; Patterson et al., 2015).”

“Different lowercase letters indicate different statistical significance by two-tailed unpaired t test: a, p < 0.05; b, p < 0.01; c, p < 0.005; and d, p < 0.001 versus AL. still the prototypical pathway and currently the most intensely studied (Robidoux et al., 2004; Susulic et al., 1995).”

“Most fasting-related studies have focused on BAT, but not subcutaneous WAT, revealing that fasting suppresses both BAT UCP1 expression and norepinephrineinduced thermogenesis (Desautels and Dulos, 1988; Hayashi and Nagasaka, 1983; Sivitz et al., 1999).”

“Gut Microbiota Orchestrates EODF-Induced WAT Beiging Recently, correlative evidence revealed a metabolic interaction between the gut microbial communities and the host, and gut bacteria have an important role in the regulation of brown and beige adipose tissues (Chevalier et al., 2015; Holmes et al., 2012; Mestdagh et…”

“The mechanism by which EODF induces higher energy expenditure might differ from that of cold exposure; cold exposure is closely tied to b-AR signaling (Cypess et al., 2015; Mund and Frishman, 2013), while fasting is known to suppress norepinephrine turnover (Knehans and Romsos, 1983) and norepinephrine-induced thermog…”

“Intriguingly, recent studies indicate that both acetate and lactate are beiging inducers (Hanatani et al., 2016; Kim et al., 2017; Sahuri-Arisoylu et al., 2016).”

“The cecal content samples for NMRwas prepared as previously described (Tian et al., 2012).”

“The residue was dissolved in methanol: H2O (80:20, v/v, pH 2-3) and injected into the GC–MS.”

“Intriguingly, recent studies indicate that both acetate and lactate are beiging inducers (Hanatani et al., 2016; Kim et al., 2017; Sahuri-Arisoylu et al., 2016).”

“, 2015; Mund and Frishman, 2013), while fasting is known to suppress norepinephrine turnover (Knehans and Romsos, 1983) and norepinephrine-induced thermogenesis in BAT (Hayashi and Nagasaka, 1983).”

“Most fasting-related studies have focused on BAT, but not subcutaneous WAT, revealing that fasting suppresses both BAT UCP1 expression and norepinephrineinduced thermogenesis (Desautels and Dulos, 1988; Hayashi and Nagasaka, 1983; Sivitz et al., 1999).”

“In addition, both BAT and WAT express protonlinked monocarboxylate transporter 1 (MCT1) encoded by Mct1, which drives acetate and lactate transport across the plasma membrane of adipocytes (den Besten et al., 2013; Iwanaga et al., 2009).”

“, 2015), for centuries and has been proven to benefit body composition and health (Eshghinia and Mohammadzadeh, 2013; López-Bueno et al., 2014; Patterson et al., 2015).”

“Most fasting-related studies have focused on BAT, but not subcutaneous WAT, revealing that fasting suppresses both BAT UCP1 expression and norepinephrineinduced thermogenesis (Desautels and Dulos, 1988; Hayashi and Nagasaka, 1983; Sivitz et al., 1999).”

“Recent studies have shown that interactions between the host and gut microbiota affect many aspects of energy metabolism (Takeda, 2016).”