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Fenugreek (Trigonella foenum-graecum) is an annual herbaceous plant and a staple of traditional health remedies for metabolic conditions including high cholesterol and diabetes. While the mechanisms of the beneficial actions of fenugreek remain unknown, a role for intestinal microbiota in metabolic homeostasis is likely. To determine if fenugreek utilizes intestinal bacteria to offset the adverse effects of high fat diets, C57BL/6J mice were fed control/low fat (CD) or high fat (HFD) diets each supplemented with or without 2% (w/w) fenugreek for 16 weeks. The effects of fenugreek and HFD on gut microbiota were comprehensively mapped and then statistically assessed in relation to effects on metrics of body weight, hyperlipidemia, and glucose tolerance. 16S metagenomic analyses revealed robust and significant effects of fenugreek on gut microbiota, with alterations in both alpha and beta diversity as well as taxonomic redistribution under both CD and HFD conditions. As previously reported, fenugreek attenuated HFD-induced hyperlipidemia and stabilized glucose tolerance without affecting body weight. Finally, fenugreek specifically reversed the dysbiotic effects of HFD on numerous taxa in a manner tightly correlated with overall metabolic function. Collectively, these data reinforce the essential link between gut microbiota and metabolic syndrome and suggest that the preservation of healthy populations of gut microbiota participates in the beneficial properties of fenugreek in the context of modern Western-style diets.
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SCIENTIFIC REPORTS | (2020) 10:1245 |
Fenugreek Counters the Eects of
High Fat Diet on Gut Microbiota in
Mice: Links to Metabolic Benet
Annadora J. Bruce-Keller1*, Allison J. Richard1, Sun-Ok Fernandez-Kim1, David M. Ribnicky2,
J. Michael Salbaum1, Susan Newman1, Richard Carmouche1 & Jacqueline M. Stephens1
Fenugreek (Trigonella foenum-graecum) is an annual herbaceous plant and a staple of traditional health
remedies for metabolic conditions including high cholesterol and diabetes. While the mechanisms
of the benecial actions of fenugreek remain unknown, a role for intestinal microbiota in metabolic
homeostasis is likely. To determine if fenugreek utilizes intestinal bacteria to oset the adverse
eects of high fat diets, C57BL/6J mice were fed control/low fat (CD) or high fat (HFD) diets each
supplemented with or without 2% (w/w) fenugreek for 16 weeks. The eects of fenugreek and HFD on
gut microbiota were comprehensively mapped and then statistically assessed in relation to eects on
metrics of body weight, hyperlipidemia, and glucose tolerance. 16S metagenomic analyses revealed
robust and signicant eects of fenugreek on gut microbiota, with alterations in both alpha and beta
diversity as well as taxonomic redistribution under both CD and HFD conditions. As previously reported,
fenugreek attenuated HFD-induced hyperlipidemia and stabilized glucose tolerance without aecting
body weight. Finally, fenugreek specically reversed the dysbiotic eects of HFD on numerous taxa
in a manner tightly correlated with overall metabolic function. Collectively, these data reinforce the
essential link between gut microbiota and metabolic syndrome and suggest that the preservation
of healthy populations of gut microbiota participates in the benecial properties of fenugreek in the
context of modern Western-style diets.
Obesity linked to Western-style diets is the prototypical ailment of the modern era. Obesity currently aects more
than 35% of Americans1; and in addition to ties with type 2 diabetes and cardiovascular disease, obesity increases
the risk of all-cause mortality and exacerbates anxiety and depression25. While search for eective obesity treat-
ments has become a priority in biomedical research, available pharmacological options for obesity are under-
mined by issues related to toxicity and o-target side6. Herbal medicine or phytotherapy has long been a source
of traditional medicinal remedies, and indeed, interest in generally regarded as safe (GRAS) plant materials for
the clinical treatment of obesity is growing (reviewed in7,8). Fenugreek (Trigonella foenum-graecum) is an annual
herbaceous plant and a staple of traditional health remedies to treat hyperlipidemia and diabetes912, as well as
mood disorders13. Laboratory studies demonstrate protective eects of fenugreek on diabetes1418, and suggest
that potential mechanisms might include inhibition of intestinal glucose absorption1416, delayed gastric empty-
ing15, and/or insulinotropic activity19,20,17,18. Protective eects of fenugreek on cholesterol and hyperlipidemia21
might be based on modulation of hepatic steatosis2226, inammation2628, and/or oxidative stress secondary to
diabetes2932. While the exact mechanisms whereby fenugreek or its constituents confers metabolic resiliency are
unknown, data show that fenugreek administration can also modulate intestinal microbiota, which can in turn
impact metabolic physiology33,34.
A remarkably mutualistic relationship exists between gut microbiota and their mammalian hosts, with micro-
biota providing protection against ingested pathogens, neutralizing carcinogens, and metabolizing otherwise
inaccessible lipids and polysaccharides into potent bioactive metabolites35. Sequencing data show that mod-
ern high fat/calorie diets can disrupt gut microbiota, reducing bacterial diversity and upsetting the balance of
pathogenic and commensal bacteria36. Data from our lab and others show that such diet-induced gut dysbiosis
is sucient to impair both metabolic and neurologic function37,38, suggesting that preservation of healthy gut
microbiota could oset the pathophysiologic eects of high fat diets39. As fenugreek has indeed been shown
1Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, 70808, USA.
2Department of Plant Biology, Rutgers University, New Brunswick, NJ, 08901, USA. *email: annadora.bruce-keller@
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SCIENTIFIC REPORTS | (2020) 10:1245 |
to modulate intestinal bacteria in several models33,34,40, studies were designed to determine if fenugreek could
oset the eects of a high fat diet on gut dysbiosis, and to establish the relationship of fenugreek-shaped gut
microbiota to clinically relevant metrics of metabolic function. To this end, data from our previously published
study on the eects of fenugreek on mice given a high fat diet were extended to include sequencing and statistical
assessment of gut microbiota. As reported in our previous study, high fat or nutritionally matched low fat diets
supplemented with ground fenugreek seeds (2% w/w) were administered to male C57BL/6J mice for 16 weeks,
and the metabolic eects of the various diets on adiposity, glycemic control, and hyperlipidemia were quantied41.
Metagenomic sequencing of fecal microbiota collected from mice was conducted, and diet-related changes in gut
microbiota were statistically analyzed in relation to established metrics of metabolic function.
Fenugreek improves glucose tolerance and dyslipidemia in mice given high fat diet. Data in
this manuscript is built on initial publication of the eects of whole fenugreek seed supplementation (2% w/w)
on overall metabolic function in the context of a 16-week trial of high fat diet consumption41, and thus previously
published data are only summarized in this report. Briey, data show that fenugreek supplementation increased
HDL and decreased LDL cholesterol levels in high fat fed-mice (Table1). Furthermore, fenugreek signicantly
improved glucose tolerance (as measured 40 minutes aer oral glucose loading), but did not aect HFD-induced
changes in total cholesterol, body weight, amount of body fat, or fasting blood glucose (Table1). Fenugreek
administration did not cause changes in food intake41.
Fenugreek and high fat diet exert pronounced effects on gut microbial composition. The
impact of fenugreek on intestinal microbiota was determined by 16S sequencing of fecal samples isolated from
mice at euthanasia as described in Methods. Initial weighted and unweighted Unifrac phylogenetic analyses
reveal that the microbiomes of fenugreek-fed mice were signicantly dierent from non-fenugreek mice under
both control diet and high fat-fed conditions (Table2). Indeed, the magnitude of the eects of fenugreek were
similar in that of the high fat dies as compared to control diet (Table2). ese robust shis in beta-diversity were
also apparent on principal component analysis plots generated from normalized read count data (Fig.1). Finally,
Total Cholesterol
(mg/dl) 148.9 ± 45.7 136.2 ± 31.7 255.4 ± 32.9*** 245.7 ± 26.3
LDL Cholesterol
(mg/dl) 9.62 ± 2.6 8.48 ± 2.1 17.98 ± 4.7*** 13.83 ± 4.3#
HDL Cholesterol
(%TC) 45.15 ± 12.7 44.86 ± 5.8 28.28 ± 3.3*** 33.3 ± 5.1#
Body Weight (gr) 31.78 ± 3.2 31.57 ± 2.6 48.73 ± 2.7*** 50.03 ± 2.2
Body Fat (gr) 5.15 ± 1.8 5.04 ± 1.2 16.61 ± 1.3*** 16.86 ± 1.3
Fasting Blood
Glucose (mg/dl) 153.4 ± 16.8 149.4 ± 23.4 211.1 ± 16.8*** 219.1 ± 21.1
Glucose Tolerance
(40 min) 234.0 ± 49.3 213.5 ± 23.3 386.1 ± 89.9*** 311.6 ± 75.8#
Table 1. Summary of fenugreek-induced metabolic resiliency: decreased hyperlipidemia and improved glucose
tolerance. Adult male C57Bl/6 mice were given high fat (HFD) or nutritionally matched control diet (CD) with
or without fenugreek (FG; 2% w/w), and subject to measures of metabolic function as described in Methods.
Statistically signicant dierences in metabolic parameters in HFD-fed mice as compared to CD-fed mice are
mice are noted by ***(p < 0.001), while signicant changes in mice given HFD/FG as compared to HFD-fed
mice are noted by #(p < 0.05). Adapted from previously published data41.
Comparison Score P value
Weighted Unifrac
CD vs CD/FG 0.65012 <0.001***
FG 0.61915 <0.001***
CD vs HFD 0.796987 <0.001***
Unweighted Unif rac
CD vs CD/FG 0.90777 <0.001***
FG 0.885073 0.002009**
CD vs HFD 0.952579 0.001009**
Table 2. Dierences in microbiota community composition in mice with CD- and HFD-shaped microbiota
with and without fenugreek. Operational taxonomical units (OTU) were identied based on sequence
clustering as described in Methods, and generation of a read count table was performed with the soware
package ‘usearch. Statistical tests for dierential representation were performed with tools incorporated in
‘mothur’, and statistically signicant dierences in microbiota community composition between groups were
detected using both weighted and unweighted Unifrac phylogenetic analysis tools.
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data show that fenugreek also signicantly increased alpha diversity (Shannon metrics) in both control diet and
high fat-fed mice (Fig.2).
Fenugreek can correct the dysbiotic effects of high fat diet on intestinal microbial popu-
lations. To assess the impact of fenugreek and HFD on gut microbial composition in greater statistical
detail, a dierential analysis of count data was conducted using DESeq. 2. Specically, the specic individual
operational taxonomic units (OTUs) whose relative representation was signicantly (p < 0.05 adjusted with
Benjamini-Hochberg correction) changed by high fat diet were identied using DESeq. 2. ese analyses revealed
that out of 410 Core OTUs (identied in all mice), the relative representation of 147 individual OTUs was sig-
nicantly dierent in HFD-fed mice as compared to CD-fed mice (Fig.3); with 57 increased and 90 decreased,
respectively, by HFD. A similar DESeq. 2 analysis of these 147 “HFD-transformed” taxa was conducted to identify
those that were signicantly aected by fenugreek such that the direction of the change induced by high fat diet
was reversed. is ivvestigation of “fenugreek-corrected” taxa revealed that fenugreek corrected the eects of
HFD on 50 of these OTUs by reducing the representation of 27 HFD-increased OTUs and augmenting the rep-
resentation of 23 OTUs reduced by HFD (Fig.3). ese analyses reect the robust eect of both fenugreek and
HFD on gut microbiota, and show that fenugreek is able to signicantly correct much (greater than 34%) of the
dysbiotic eects of HFD.
Representation of Fenugreek-corrected taxa largely predicts overall metabolic function. In
the nal set of analyses, the 50 fenugreek-corrected taxa whose representation was skewed in one direction by
HFD but in the opposite direction by fenugreek was examined in relation to metabolic function. Specically, to
determine if the relative representation of fenugreek-corrected taxa could be used to predict metabolic resiliency,
the statistical relationship of fenugreek-corrected taxa representation to the metrics of metabolic function listed in
Table1 was determined. To this end, a matrix was built containing OTU count data for all 50 fenugreek-corrected
Figure 1. Fenugreek changes intestinal microbial populations in mice. Fecal microbiome populations from CD,
CD/FG, HFD, and HFD/FG mice were analyzed using 16S rRNA sequencing, and multi-dimensional scaled
principal coordinate analysis were used to visualize UniFrac distances of fecal samples from individual recipient
mice. Samples from CD, CD/FG, HFD, and HFD/FG mice are depicted as blue, purple, red, and green symbols,
Figure 2. Fenugreek increases overall intestinal microbial diversity. Fecal microbiome populations from CD,
CD/FG, HFD, and HFD/FG mice were analyzed using 16S rRNA sequencing, and box plots were generated to
depict dierences in Shannon α-diversity. Data show that mice supplemented with 2% fenugreek in their feed
exhibited a statistically signicant (**p < 0.01) increase in α-diversity compared to mice given CD or HFD
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SCIENTIFIC REPORTS | (2020) 10:1245 |
taxa along with all metabolic data depicted in Table1, including those indices not aected by fenugreek. Pairwise
Pearson correlations indicate that the relative expression of many of these 50 OTUs were highly predictive of met-
abolic function. For example, of the 23 taxa decreased by HFD but increased with fenugreek supplementation, 8
taxa (all in the fermicutes phylum) signicantly correlated with at least 1 metric of metabolic function (Table3,
see Supplementary Table1 for additional details (log2FC and Pearson r values) on HFD-decreased, fenugreek
corrected taxa). Likewise, of the 27 taxa increased by HFD but decreased with fenugreek supplementation, 20
taxa signicantly correlated with selected metrics of metabolic function (Table4, see Supplementary Table2 for
additional details (log2FC and Pearson r values) on HFD-increased, fenugreek corrected taxa). It is important
to note that the representation of fenugreek-corrected taxa correlated frequently with aspects of metabolic func-
tion (e.g., body weight, body fat, total cholesterol, fasting blood glucose) that were not signicantly improved in
fenugreek-treated mice.
While benecial eects of fenugreek on hyperlipidemia and hyperglycemia have been widely reported, the mech-
anisms of fenugreek-mediated actions on metabolic function are unknown. Here we demonstrate the robust
eects of fenugreek on gut microbiota, and describe a novel combination of statistical analyses including Unifrac,
iterative DESeq. 2, and pairwise correlation matrices to generate insight into the role of intestinal microbial
changes in the protective eects of fenugreek. Specically, sequencing analyses reveal that fenugreek signicantly
increased overall microbiome diversity in mice, and specically reversed the actions of high dietary fat on key
intestinal taxa. Furthermore, the representation of fenugreek-corrected taxa signicantly correlated with meta-
bolic function, including changes in body weight and composition, glucose regulation, and hyperlipidemia.ese
ndings are in agreement with the extensive body of literature documenting the ability of high fat diet to reduce
bacterial diversity and disrupt the balance of pathogenic/commensal bacteria within the intestine36,42,38,43. As data
from our lab and others show that this pattern of gut dysbiosis is sucient to impair both metabolic and neuro-
logic function37,4448, data in this paper suggest that the reported eects of fenugreek on gut microbiota33,34,40,49,
may be fundamental to its benecial properties. In light of the stubborn prevalence of obesity and the pervasive
accessibility of unhealthy diets, it is both clinically signicant and generally promising that these data suggest
that partial reversal of gut dysbiosis and metabolic impairment can be achieved with botanical supplementation
within the context of unhealthy, Western-style diets.
While the association of intestinal dysbiosis with metabolic disease is well established4548 causal relationships
have not been identied and it is not understood how microbiome constituents impact metabolic resilience/vul-
nerability. ree major phyla are the most abundant in the human distal intestine: Bacteroidetes (gram-negative),
Firmicutes (gram-positive), and Actinobacteria (gram-positive). Early studies suggested that obesity causes
reductions in Bacteroidetes and increases in Firmicutes38,43 and indeed further studies indicate that these changes
are reversed with weight loss50,51. While these data suggest that the balance between these phyla might broadly
impact host physiology, this binary distinction does not always occur52,53 and may be too simple to faithfully
reect the complexity of diet-induced changes to the gut microbiome5456. In the present study as well as our
previous studies54,57, dierences between control and high fat groups did not manifest as phylum-level shis
but rather dierential representation within taxa, particularly Firmicutes. Indeed, the majority of taxa included
in Tables3 and 4 arise from the Clostridium class (Clostridium cluster XIVa) or the Clostridium leptum group
(Clostridium cluster IV). is is notable, as divergent shis in the representation of Clostridia have been reported
in other pathophysiological conditions. For example, while overall Clostridium representation generally increases
with age, Clostridium XIVa clusters have been shown to be signicantly reduced in the elderly58. e bacteria
in Clostridium XIVa play major roles in the fermentation of carbohydrates within the gut59, and the major end
products of hind-gut fermentation are short-chain fatty acids (SCFAs). Further, Fermicutes produce primarily
butyrate as their metabolic end product60, and butyrate is the main source of nutrition for gut epithelium cells61.
Depletion of butyrate is associated with impaired intestinal barrier integrity62, and loss of intestinal barrier func-
tion is in turn associated with a growing number of inammatory disease states diseases, including obesity as well
as autoimmune diseases and cancer (reviewed in63,64). While butyrate was not directly measured in the present
Figure 3. Graphical representation of fenugreek’s ability to reverse HFD-induced changes in individual
intestinal microbiota. e ability of high fat diet to signicantly alter the representation of individual taxa was
assessed as described in Methods showing that out of 410 Core OTUs, 57 were signicantly increased and 90
signicantly decreased by HFD. Fenugreek supplementation signicantly reversed the eects of HFD on 50 of
these OTUs by reducing the representation of 27 HFD-increased OTUs and bolstering the representation of 23
OTUs reduced by HFD.
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study, decreased levels of butyrate and other SCFA are widely reported in the context of obesity while high-ber
plant products are known to increase colonic fermentation and the generation of SCFA65.
Our data indicate that fenugreek is particularly eective against hyperlipidemia, which is in keeping with
results of both experimental and clinical studies14,22,30,66. While there are several mechanisms whereby changes
in gut microbiota could mediate the eects of fenugreek on serum lipids, most ultimately impact the absorp-
tion of dietary fat. For example, intestinal microbiota alter the metabolism of diet-derived long-chain fatty acids
such as conjugated linoleic acid, modulating absorption67. Gut microbiota can also moderate cholesterolemia
Correlation (Pearson) of HFD-Decreased, Fenugreek-Corrected OTU’s to Metrics of Metabolic Function
Individual OTUs Decreased by High Fat Diet (HF) Corrected by FG Total
Chol. LDL
Chol. HDLChol. Body
Wei g ht Bod y Fat Fasting
Glucose Glucose
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species1 ns ns ns ns 0.0014 ns 0.0001
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species2 ns ns ns 0.0027 0.0025 ns ns
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species3 0.0001 0.0034 0.0011 ns ns 0.0006 0.0001
Firmicutes/Clostridia/Clostridiales/Ruminococcaceae/Flavonifractor 1.2E-05 0.0005 0.0001 1.6E-06 2.1E-06 2.2E-05 1.9E-05
Firmicutes/Erysipelotrichia/Erysipelotrichales/Erysipelotrichaceae/Turicibacter 0.0004 ns 0.0024 ns ns ns 0.0015
Firmicutes/Clostridia/Clostridiales/Ruminococcaceae/Oscillibacter 0.0002 ns 1.3E-05 ns 4.0E-05 0.0002 4.2E-05
Firmicutes/Clostridia/Clostridiales/Ruminococcaceae/Intestinimonas 0.0009 0.0021 0.0026 ns ns 0.0004 0.0017
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Acetatifactor/Species 1 ns ns ns ns ns ns 0.0012
Table 3. OTUs that predicts metabolic decline in high fat-fed mice. Individual OTUs in which fenugreek
administration reversed high fat diet-induced decreases in representation were correlated against measures of
hyperlipidemia. Data are p values of Pearson correlation with total cholesterol (mg/dl), low-density lipoprotein
(LDL Chol.; mg/dl), high-density lipoprotein (HDL Chol.; %TC), body weight (grams), body fat (grams),
fasting blood glucose (mg/dl), and glucose tolerance (blood glucose levels 40 minutes aer oral loading). See
Supplementary Table1 for additional details (log2FC and Pearson r values) on HFD-decreased, fenugreek
corrected taxa.
Correlation (Pearson) of HFD-Increased, Fenugreek-Corrected OTU’s to Metrics of Metabolic Function
Individual OTUsDecreased by High Fat Diet (HF) Corrected by FG Total
Chol. LDL
Chol. HDL
Chol. Body
Wei g ht Bod y Fat Fasting
Glucose Glucose
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species 4 ns 4.9E-05 ns ns ns ns ns
Firmicutes/ClostridiaClostridiales/Ruminococcaceae/Anaerotruncus 0.0004 1.3E-08 ns ns 0.0016 ns 4.6E-06
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species 5 ns 5.8E-05 0.0024 ns ns ns ns
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species 6 0.0002 4.9E-09 0.0051 ns ns ns 0.0002
Bacteroidetes/Bacteroidia/Bacteroidales/Porphyromonadaceae/Barnesiella/Species 1 9.1E-05 4.1E-10 ns 0.0009 0.0009 0.0024 1.6E-06
Bacteroidetes/Bacteroidia/Bacteroidales/Porphyromonadaceae/Barnesiella/Species 2 0.0005 4.2E-09 ns ns ns ns 8.8E-06
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species 7 8.1E-07 5.7E-10 4.8E-05 ns ns ns 0.0001
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species 8 0.0001 0.0002 0.0030 ns 1.2E-05 0.0011 1.3E-05
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species 9 4.1E-06 5.9E-08 0.0007 ns 1.2E-06 4.1E05 ns
Firmicutes/Bacilli/Lactobacillales/Streptococcaceae/Streptococcus ns ns ns ns ns ns 0.0007
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species 10 0.0009 2.1E-05 ns ns 0.0002 ns 5.0E-06
Firmicutes/Bacilli/Lactobacillales/Lactobacillaceae/Lactobacil lus/Species 1 4.2E-06 ns ns 7.1E-08 4.3E-09 5.6E07 1.1E-06
Firmicutes/Bacilli/Lactobacillales/Lactobacillaceae/Lactobacil lus/Species 2 1.3E-06 0.0001 ns 2.9E-08 2.1E-09 2.9E-07 5.2E-07
Actinobacteria/Actinobacteria/Coriobacteridae/Coriobacteriales/Coriobacterineae ns ns ns ns ns ns 0.0033
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Roseburia 6.0E-05 5.4E-07 0.0004 ns ns ns ns
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species 11 ns ns ns 0.0030 0.0016 ns 0.0015
Firmicutes/Clostridia/Clostridiales/Clostridiales_Incertae_Sedis_XI/Dethiosulfatibacter 0.0039 ns ns 0.0004 ns ns
Bacteroidetes/Bacteroidia/Bacteroidales/Porphyromonadaceae/Barnesiella/Species 3 ns 1.4E-05 ns ns ns ns 0.0040
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Acetatifactor/Species 2 0.0003 0.0002 ns ns 1.6E-05 ns ns
Firmicutes/Clostridia/Clostridiales/Lachnospiraceae/Clostridium_XlVa/Species 12 5.2E-05 2.2E-07 0.0003 ns ns ns ns
Table 4. OTUs that predicts metabolic decline in high fat-fed mice. Individual OTUs in which fenugreek
administration reversed high fat diet-induced increases in representation were correlated against measures of
hyperlipidemia. Data are correlation coecients (Pearson r) and p values of correlation with total cholesterol
(mg/dl), low-density lipoprotein (LDL Chol.; mg/dl), high-density lipoprotein (HDL Chol.; %TC), body weight
(grams), body fat (grams), fasting blood glucose (mg/dl), and glucose tolerance (blood glucose levels 40 minutes
aer oral loading). See Supplementary Table2 for additional details (log2FC and Pearson r values) on HFD-
increased, fenugreek corrected taxa.
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SCIENTIFIC REPORTS | (2020) 10:1245 |
by regulating cholesterol conversion into coprostanol68. Dietary cholesterol is largely incorporated into chylo-
microns for absorption in the small intestine. However, signicant quantities of cholesterol (as much as 1 gram
per day) escape proximal absorption to enter the colon to be either be excreted or absorbed. Microbial-based
metabolism of cholesterol into coprostanone/coprostanol reduces blood cholesterol by increasing fecal choles-
terol excretion69,70. Interestingly, recent data suggest that taxa arising from Lachnospiraceae and Runinococcacea
families of the phylum Fermicutes are uniquely associated with high coprostanol generation in healthy humans71,
while other studies likewise link these gut microbiota to variation in blood lipid levels independently of age, sex,
and host genetics72. It is important to note that seven of the eight of the taxa listed in Table3 are members of
Lachnospiraceae and Runinococcacea families, suggesting that reversal of HFD-induced decreases in these key
taxa by fenugreek could remediate hyperlipidemia by promoting fecal fat excretion. In further support of this
scenario, published data suggest that fenugreek supplementation can dose-dependently increase fecal excretion of
cholesterol from rats given high fat/high calorie diets73. Collectively, these data raise the possibility that increased
representation of coprostanoligenic taxa arising from Lachnospiraceae and Runinococcacea families could par-
ticipate in the lipid-lowering eects of fenugreek, and further suggest that identication and analysis such strains
could lead to improved understanding and management of hypocholesteremia.
Transformation and metabolism of bile acids is another key pathway whereby fenugreek-shaped intestinal
bacteria could impact serum lipids (reviewed in74). Indeed, fenugreek has been reported to inhibit the intestinal
absorption of primary and secondary bile acids75, and to increase bile acid excretion into feces30. While the bulk of
bile acids released into the intestine are eciently absorbed and recycled back to the liver, perhaps 5% of the total
bile acid pool progresses into the colon. Bile acids reaching the colon are subject to several microbial-mediated
reactions including transformation into secondary bile acids by dihydroxylation, and deconjugation by bile
salt hydrolases. While data indicate that bile salt hydrolases are a pervasive microbial adaptation to the human
gut environment with enrichment in major genera including Bacteroides, Clostridium, Lactobacillus, and
Eubacterium76, probiotics with bile salt hydrolytic activity can lower serum cholesterol77,78. With regard to more
broad metabolic benets, secondary bile acids generated by microbial metabolism are potent ligands of the
G-protein coupled receptor TGR5, the activation of which triggers release of GLP-1 and insulin, thereby modu-
lating host glucose tolerance and energy expenditure79.
While this study is in keeping with an extensive body of literature on the benecial eects of fenugreek, the use
of whole seed supplementation precludes identication of the bioactive constituent(s) mediating changes to gut
microbiota. Furthermore, only male mice were used, so sex-based dierences in the eects of fenugreek or the
relation of such to metabolic function cannot be resolved. is point is especially signicant as female C57BL/6J
mice are generally considered resistant to diet induced obesity80,81. Notwithstanding these limitations, these data
indicate that fenugreek supplementation can stabilize metabolic function within the context of high fat consump-
tion. Indeed, fenugreek did not aect food intake or alleviate diet-induced obesity, but rather was able to bolster
resistance of the obese mice to hyperlipidemia and glucose intolerance. We use the term “metabolic resiliency” to
describe this ability to preserve, at least in part, a healthy metabolic phenotype in the context of powerful external
stressors – in this case sustained consumption of a high fat diet and the obese state. is is a signicant nding,
as while the components of a healthy lifestyle are generally well known, numerous societal factors including
poverty, food deserts, irregular/sedentary work schedules combine to hinder a consistently healthy lifestyle for
most Americans. us, use of fenugreek and/or other strategies to maintain a healthy population of intestinal
microbes in the context of a high fat diet could foster metabolic resilience even when diets/lifestyles are not opti-
mal. Indeed, while fenugreek conferred protection against hyperlipidemia and glucose intolerance, it is important
to note that representation of specic fenugreek-corrected taxa correlated frequently with aspects of metabolic
function (e.g., body weight, body fat, total cholesterol, fasting blood glucose) that were not signicantly improved
in fenugreek-treated mice (see Tables3 and 4). While correlation does not equal causation, these data clearly
illustrate the very close relationship of individual microbes and microbial balance with metabolic function, and
raise the possibility that strategic manipulation of key intestinal taxa could result in a more complete reversal of
the adverse action of high fat diet. us, the action of fenugreek could be possibly optimized by dose, preparation,
or combination with additional factors (probiotics, etc) to have a greater eect on key microbiota, enhancing
its benecial prole. ese data also suggest that perhaps intestinal microbial “ngerprints” could be generated
to estimate vulnerability to metabolic dysfunction and/or the potential for ecacy of metabolic interventions.
Overall, data in the manuscript strongly suggest that the development of microbially-targeted therapies – both
primary and adjunctive – that are built upon safe, natural, plant-based products like fenugreek could be used to
attain signicant advancements in public health within the context of contemporary dietaryenvironments.
Materials and Methods
Animals and diets. is study was carried out in strict accordance with PHS/NIH guidelines on the use of
experimental animals, and all experimental protocols were approved by the Institutional Animal Care and Use
Committee at Pennington Biomedical Research Center. Data in this manuscript follows an initial publication on
the eects of whole fenugreek seed supplementation (2% w/w) on overall metabolic function in the context of a
16-week trial of high fat diet consumption41. As detailed in our initial report41, male, 9 week-old C57BL/6 J mice
were purchased from Jackson Laboratories, and group-housed (4/cage) under standard conditions with ad libi-
tum access to food/water. Aer 7 days acclimation, mice were randomly separated into the following 4 groups (20
mice each in each group): high fat diet ± fenugreek (HFD and HFD/FG) and control diet ± fenugreek (CD and
CD/FG) for 16 weeks. HFD and HFD/FG mice were fed a diet with 60% kcal from fat without or with 2% fenu-
greek seed powder incorporated into the diets, respectively (Research Diets Inc. D12492, D16020410), while CD
mice were fed a nutritionally matched control low fat diet (10% kcal from fat) with or without 2% fenugreek seed
powder (Research Diet Inc.; D12450J, D16020408). All diets contained 10% kcal from protein with the balance
in caloric intake provided by dierences in carbohydrate content. T. foenum-graecum L. “Fenugreek” seeds were
Content courtesy of Springer Nature, terms of use apply. Rights reserved
SCIENTIFIC REPORTS | (2020) 10:1245 |
purchased from Johnny’s Selected Seeds, Winslow Maine, certied organic for sprouting. Fenugreek seeds were
ground in the Department of Plant Biology at Rutgers University and hand-delivered to Research Diets Inc., (New
Brunswick, NJ) for commercial incorporation into treatment diets at 2% of the diet by weight.
Metabolic phenotyping. Assessment of metabolic function is fully detailed in our previous report41. Briey,
body composition (fat mass, fat-free/lean mass, and water content) was measured by briey placing in mice into
Bruker minispec LF110 time domain NMR analyzer (Bruker Optics, Billerica MA) as described previously41.
Glucose tolerance was measured using an oral glucose tolerance assay (OGTT) based on repeated sampling of tail
blood using a glucometer (Ascensia Elite, Bayer, Mishawaka, IN) at 0, 20, 40, 60, and 120 minutes aer oral glu-
cose (2 gm/kg) administration. All mice remained in the study for the duration of the 16-week feeding trail, aer
which mice with euthanized following a 8-hr fast by decapitation under deep isourane anesthesia. Levels of total
cholesterol, HDL cholesterol, LDL cholesterol, and triglycerides in serum collected at euthanasia were measured
colorimetrically (Wako Chemicals, Richmond, VA).
16S Metagenomic sequencing. Fecal samples were collected at euthanasia, and DNA preparation,
sequencing and bioinformatics were performed by the PBRC Genomics Core Facility. DNA was isolated
using a commercial reagent system (MoBio Power Fecal Kit, MoBio Laboratories, Carlsbad, CA) augmented
by enzymatic lysis using lysostaphin, mutanolysin, and lysozyme82. Sequencing libraries targeting V4 of the
gene encoding the 16S ribosomal RNA were generated using a commercially available kit (NEXTex 16S V4
Amplicon-Seq Library Prep Kit, BIOO Scientic, Austin, TX), relying on 16S gene-specic primer sequences V4F
adaptors and molecular barcodes as described by the manufacturer to produce 253 bp amplicons. Samples were
sequenced with custom primers (BIOO Scientic, Austin, TX) on an Illumina MiSeq instrument using version
3 sequencing chemistry (300 bp paired end reads). Forward and reverse sequence reads were processed into
double-stranded DNA contigs using quality control metrics implemented in the soware package ‘mothur’83.
Sequence clustering (at better than 97% identity) to identify operational taxonomical units (OTUs), removal
of chimeric sequences, and generation of a read count table (i.e. tabulating the occurrence of each OTU in each
sample) were performed with the soware package ‘usearch84. Taxonomical classication of each OTU sequence
relied on the SILVA 16S rRNA sequence database version 123.185, and statistical tests for dierential representa-
tion were performed with tools incorporated in ‘mothur’, as well as using the soware package DESeq. 286.
Relative abundance of each OTU was examined on the phylum, class, order, family and genus levels.
Statistical analyses. Biochemical data were analyzed using Prism soware (GraphPad Soware, Inc.), and
displayed as mean ± standard error, and were analyzed by ANOVA. Statistical signicance for all analyses was
accepted at p < 0.05, and *, **, and *** represent p < 0.05, p < 0.01, and p < 0.001, respectively. Alpha diver-
sity (chao1 metrics) and beta diversity (weighted UniFrac metrics87) were assessed using tools implemented in
‘mothur’ on the basis of 80,000 sequences per sample. Dierential representation of OTUs was assessed using
DESeq. 2 on the basis of sequence count data, relying on Wald statistics with Benjamini-Hochberg correction
and a false discovery rate cuto set at 0.1. Inter-sample relationships relying on Principal Component Analysis
on the basis of DESeq. 2 output, and data visualizations were both performed using JMP Genomics soware
(SAS, Cary, NC). Pairwise Pearson correlations of individual metrics of metabolic function against individual
HFD-transformed, fenugreek-corrected OTU expression were carried out using Prism soware.
Received: 12 July 2019; Accepted: 7 January 2020;
Published: xx xx xxxx
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e authors gratefully acknowledge Dr. Kem Singletary and Ms. Cynthia Kloster for expert veterinary assistance
and insight into the described studies. ese studies were supported by grants from the National Center For
Complementary & Integrative Health and the Office of Dietary Supplements of the National Institutes of
Health (RO1AT010279 and P50AT002776; which funds the Botanical Dietary Supplements Research Center
of Pennington Biomedical Research Center and the Department of Plant Biology and Pathology in the School
of Environmental and Biological Sciences (SEBS) of Rutgers University). These studies also utilized the
facilities of the Animal Behavior/Phenotyping and Genomics Cores that are supported in part by COBRE (NIH
P20-GM103528) and NORC (NIH P30-DK072476) center grants from the National Institutes of Health.
Author contributions
A.B.K. and J.M.S. conceived the experiments. D.M.R. procured and performed all quality control on the fenugreek
seeds. S.F.K., A.J.R., and A.B.K. conducted the animal experiments. R.C. performed metagenomic sequencing,
while S.N., J.M.S., and A.B.K. analyzed the data. A.B.K. and J.M.S. prepared the manuscript and gures. All
authors reviewed the manuscript.
Competing interests
e authors declare no competing interests.
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... Recently, many studies reported Antifungal properties (Sudan et al., 2020), as well as effficacy and safety of fenugreek Seeds on treatment of testosterone deficiency syndrome (Mansoori et al., 2020;Park et al., 2019). Similarly Syed et al. (2020), reported nutrition and therapeutic effect of fenugreek while, Bruce-keller et al. (2020), reported that fenugreek ha ability to counter the effects of high fat diet on gut microbiota in mice. Similarity, Kaur and Sadwa (2020), reported the phytomodulatory potential of fenugreek (Trigonella foenum-graecum) on bisphenol-A induced testicular damage in mice. ...
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Background Gentamicin is one of aminoglycoside antibiotic used for treatment of many infections due to its availability and less cost. The aim of this study aimed to assess the modulation effect of fenugreek seed and its germinated seeds on pancreatic and testicular toxicity induced by gentamicin in male Swiss albino mice. Forty male albino mice were divided into four treatment groups as follows: (1) control group, (2) gentamicin treated group, (3) gentamicin-fenugreek treated group and (4) gentamicin-germinated fenugreek treated group. Pancreatic and testicular tissues were collected for histopathological examinations, histochemical, and biochemical analysis as well as genetic study. Results Administration of gentamicin resulted in histopathological damage in pancreatic and testicular tissues as well as decreased glutathione peroxides, catalase and total antioxidant activity content in both pancreatic and testicular tissues compared to control group. Histopathological changes and antioxidant/oxidative alterations as well as DNA damage observed in gentamicin treated animals found were moderate improvement by fenugreek seeds administration and marked improvement by treatment with germinated fenugreek seeds. Conclusions Treated with gentamicin induced histopathological lesions, antioxidant/oxidant imbalance and DNA damage in the pancreatic and testicular. Treatment with germinated fenugreek seeds was more effective than fenugreek seeds in amelioration of pancreatic and testicular lesions, preventing high appearance of carbohydrate and accumulation of collagen fibers as well as oxidative damage and genotoxicity induced by gentamicin administration.
... Murine studies have also reported the therapeutic relevance of fenugreek fiber in T2D by improving glucose tolerance, dyslipidemia, and weight gain. In addition, fenugreek fiber also enhances the abundance of phylum Bacteroidetes and family Lachnospiraceae, members of which are often depleted among obese people and post-menopausal T2D patients (Bruce-Keller et al. 2020;Ozaki et al. 2021;Shtriker et al. 2018). Lastly, clinical studies have proven that fenugreek seed is effective against hyperlipidemia, inflammation, and oxidative stress in T2D patients. ...
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Type 2 diabetes (T2D) and T2D-associated comorbidities, such as obesity, are serious universally prevalent health issues among post-menopausal women. Menopause is an unavoidable condition characterized by the depletion of estrogen, a gonadotropic hormone responsible for secondary sexual characteristics in women. In addition to sexual dimorphism, estrogen also participates in glucose–lipid homeostasis, and estrogen depletion is associated with insulin resistance in the female body. Estrogen level in the gut also regulates the microbiota composition, and even conjugated estrogen is actively metabolized by the estrobolome to maintain insulin levels. Moreover, post-menopausal gut microbiota is different from the pre-menopausal gut microbiota, as it is less diverse and lacks the mucolytic Akkermansia and short-chain fatty acid (SCFA) producers such as Faecalibacterium and Roseburia. Through various metabolites (SCFAs, secondary bile acid, and serotonin), the gut microbiota plays a significant role in regulating glucose homeostasis, oxidative stress, and T2D-associated pro-inflammatory cytokines (IL-1, IL-6). While gut dysbiosis is common among post-menopausal women, dietary interventions such as probiotics, prebiotics, and synbiotics can ease post-menopausal gut dysbiosis. The objective of this review is to understand the relationship between post-menopausal gut dysbiosis and T2D-associated factors. Additionally, the study also provided dietary recommendations to avoid T2D progression among post-menopausal women. Keywords: Dietary regulation, Dysbiosis, Estrogen deficiency, Gut microbiota, Post-menopause, Type 2 Diabetes
... Trigonella foenum-graecum L. and Polygonatum sibiricum Redoutè are medicinal plants with effects on mental health that contain substantial amounts of steroidal saponins. T. foenum-graecum substantially corrected the dysbiotic effect of a high-fat diet (HFD) in mice, especially regarding the Firmicutes phylum [177]. The addition of T. foenum-graecum to feed positively influenced the gut microbiome composition and immune parameters in weaning piglets [178], and in cultivation-based plate count assays, a saponin-rich P. sibiricum extract increased the abundance of probiotic bacteria and decreased the abundance of potentially harmful species [164]. ...
... Trigonella foenum-graecum L. and Polygonatum sibiricum Redoutè are medicinal plants with effects on mental health that contain substantial amounts of steroidal saponins. T. foenum-graecum substantially corrected the dysbiotic effect of a high-fat diet (HFD) in mice, especially regarding the Firmicutes phylum [177]. The addition of T. foenum-graecum to feed positively influenced the gut microbiome composition and immune parameters in weaning piglets [178], and in cultivation-based plate count assays, a saponin-rich P. sibiricum extract increased the abundance of probiotic bacteria and decreased the abundance of potentially harmful species [164]. ...
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Background: Various neurocognitive and mental health-related conditions have been associated with the gut microbiome, implicating a microbiome-gut-brain axis (MGBA). The aim of this systematic review was to identify, categorize, and review clinical evidence supporting medicinal plants for the treatment of mental disorders and studies on their interactions with the gut microbiota. Methods: This review included medicinal plants for which clinical studies on depression, sleeping disorders, anxiety, or cognitive dysfunction as well as scientific evidence of interaction with the gut microbiome were available. The studies were reported using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. Results: Eighty-five studies met the inclusion criteria and covered thirty mental health-related medicinal plants with data on interaction with the gut microbiome. Conclusion: Only a few studies have been specifically designed to assess how herbal preparations affect MGBA-related targets or pathways. However, many studies provide hints of a possible interaction with the MGBA, such as an increased abundance of health-beneficial microorganisms, anti-inflammatory effects, or MGBA-related pathway effects by gut microbial metabolites. Data for Panax ginseng, Schisandra chinensis, and Salvia rosmarinus indicate that the interaction of their constituents with the gut microbiota could mediate mental health benefits. Studies specifically assessing the effects on MGBA-related pathways are still required for most medicinal plants.
The beneficial effects of Lactobacillus plantarum LP104 was investigated on lipid level and the disordered gut microbiota of HFD feeding C57BL/6N mice. C57BL/6N mice were divided into three groups, ordinary diet (Control), high-fat diet (HFD) and HFD with LP104, respectively. After 8 weeks, LP104 alleviated the body weight, liver index, colon length, TC, TG, LDL, TNFα and LPS levels. High-throughput sequencing analysis has found that long-term HFD destroy microbial abundance and composition in the gut. Nevertheless, dietary supplementation of LP104 reduced the ratio of Firmicutes to Bacteroides and regulate the gut microbiota community structure positively. Meanwhile, the intake of LP104 also increased the abundance of beneficial bacteria Bifidobacterium in the cecum, which improves the intestinal flora disorder induced by a high-fat diet. The present study shows that LP104 supplementation improved the amelioration HFD induced hyperlipidemia by positively regulating intestinal microorganisms.
Fenugreek (Trigonella foenum-graecum), known as methi in much of South Asia, is a widely used spice and vegetable crop. Fenugreek is a multiuse legume crop grown in dry and semiarid regions of the developing world. It is an annual, dicotyledonous, self-pollinated plant belonging to the family Fabaceae. It is a diploid with 2n = 16 and is estimated to have 685 Mbp of genome size. The genus Trigonella L. includes about 135 species worldwide and is native to South-Eastern Europe and West Asia. Most widely known as a spice in Europe, it is also widely used medicinally and as a green vegetable or sprout and as a forage crop. Surprisingly it does not yet have a published genome, and refocusing attention on its uses may stimulate much-needed research on this underutilized crop. Key questions a genome will help address are trade-offs in performance among the various uses of fenugreek and improved understanding of its unique secondary metabolite profile.KeywordsSpice domesticationForage cropFood securityNutrition
Diabetes mellitus comprises a group of heterogeneous disorders, which are usually subdivided into type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM). Both genetic and environmental factors have been implicated in the onset of diabetes. Type 1 diabetes primarily involves autoimmune insulin deficiency. In comparison, type 2 diabetes is contributed by the pathological state of insulin deficiency and insulin resistance. In recent years, significant differences were found in the abundance of microflora, intestinal barrier, and intestinal metabolites in diabetic subjects when compared to normal subjects. To further understand the relationship between diabetes mellitus and intestinal flora, this paper summarizes the interaction mechanism between diabetes mellitus and intestinal flora. Furthermore, the natural compounds found to treat diabetes through intestinal flora were classified and summarized. This review is expected to provide a valuable resource for the development of new diabetic drugs and the applications of natural compounds.
The Chemistry inside Spices and Herbs: Research and Development brings comprehensive information about the chemistry of spices and herbs with a focus on recent research in this field. The book is an extensive 2-part collection of 20 chapters contributed by experts in phytochemistry with the aim to give the reader deep knowledge about phytochemical constituents in herbal plants and their benefits. The contents include reviews on the biochemistry and biotechnology of spices and herbs, herbal medicines, biologically active compounds and their role in therapeutics among other topics. Chapters which highlight natural drugs and their role in different diseases and special plants of clinical significance are also included. Part II continues from the previous part with chapters on the treatment of skin diseases and oral problems. This part focuses on clinically important herbs such as turmeric, fenugreek, ashwagandha (Indian winter cherry), basil, Terminalia chebula (black myrobalan). In terms of phytochemicals, this part presents chapters that cover resveratrol, piperine and circumin.
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The purpose of this experiment was to study the effects of fenugreek seed extract (FSE) on the growth performance, intestinal morphology, intestinal immunity and cecal micro-organisms in yellow-feathered broilers. A total of 240 one-day-old male yellow-feathered broilers were selected and randomly assigned to four treatments with six replicates per group and ten broilers per replicate. Started from the 3rd day, birds were fed with basal diet (CON group) or basal diet supplemented with 30 mg/kg Zinc bacitracin (ZB group), or basal diet supplemented with 50 (D-FSE group) or 100(H-FSE group) mg/kg FSE, respectively. The experiment lasted for 56 days. The results showed that dietary FSE supplementation improved average daily weight gain (ADG) and ratio of feed to weight gain (F: G) (P<0.01), increased intestinal villus height (VH), and villus height to crypt depth ratio (V/C) (P<0.05), serum concentrations of IL-10 and the contents of secretory immunoglobulin A (sIgA) (P<0.05), and decreased the activity of iNOS(P<0.05). The high-throughput sequencing results showed that dietaryFSEsupplementationincreased the alpha diversity of cecal microbes, and Firmicutes, Bacteroidetes, Verrucomicrobia and Proteobacteria taken up 95% of all phyla detected, FSE significantly reduced Campylobacter, Synergistes and Lachnoclostridium abundance (P≤0.05). There were significant difference in more than 30 KEGG pathways between FSE added group and control group or ZB group.FSE supplementation, in other words, maintained gut microbiotahomeostasis while improving broiler growth performance. As a result, FSE has the potential to replace prophylactic antibiotic use in poultry production system.
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Recently, the gut microbiota has emerged as a crucial factor that influences cholesterol metabolism. Ever since, significant interest has been shown in investigating these host-microbiome interactions to uncover microbiome-mediated functions on cholesterol and bile acid (BA) metabolism. Indeed, changes in gut microbiota composition and, hence, its derived metabolites have been previously reported to subsequently impact the metabolic processes and have been linked to several diseases. In this context, associations between a disrupted gut microbiome, impaired BA metabolism, and cholesterol dysregulation have been highlighted. Extensive advances in metagenomic and metabolomic studies in this field have allowed us to further our understanding of the role of intestinal bacteria in metabolic health and disease. However, only a few have provided mechanistic insights into their impact on cholesterol metabolism. Identifying the myriad functions and interactions of these bacteria to maintain cholesterol homeostasis remain an important challenge in such a field of research. In this review, we discuss the impact of gut microbiota on cholesterol metabolism, its association with disease settings, and the potential of modulating gut microbiota as a promising therapeutic target to lower hypercholesterolemia.
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Increased dietary fiber consumption has been associated with many beneficial effects, including amelioration of obesity and insulin resistance. These effects may be due to the increased production of short chain fatty acids, including propionate, acetate and butyrate, during fermentation of the dietary fiber in the colon. Indeed, oral and dietary supplementation of butyrate alone has been shown to prevent high fat-diet induced obesity and insulin resistance. This review focuses on sources of short chain fatty acids, with emphasis on sources of butyrate, mechanisms of fiber and butyrate metabolism in the gut and its protective effects on colon cancer and the peripheral effects of butyrate supplementation in peripheral tissues in the prevention and reversal of obesity and insulin resistance.
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To assess the metabolically beneficial effects of fenugreek (Trigonella foenum-graecum), C57BL/6J mice were fed a low- or high-fat diet for 16 weeks with or without 2% (w/w) fenugreek supplementation. Body weight, body composition, energy expenditure, food intake, and insulin/glucose tolerance were measured regularly, and tissues were collected for histological and biochemical analysis after 16 weeks of diet exposure. Fenugreek did not alter body weight, fat mass, or food intake in either group, but did transiently improve glucose tolerance in high fat-fed mice. Fenugreek also significantly improved high-density lipoprotein to low-density lipoprotein ratios in high fat-fed mice without affecting circulating total cholesterol, triglycerides, or glycerol levels. Fenugreek decreased hepatic expression of fatty acid-binding protein 4 and increased subcutaneous inguinal adipose tissue expression of adiponectin, but did not prevent hepatic steatosis. Notably, fenugreek was not as effective at improving glucose tolerance as was four days of voluntary wheel running. Overall, our results demonstrate that fenugreek promotes metabolic resiliency via significant and selected effects on glucose regulation, hyperlipidemia, and adipose pathology; but may not be as effective as behavioral modifications at preventing the adverse metabolic consequences of a high fat diet.
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Obesity is a global epidemic that has shown a steady increase in morbimortality indicators; it is considered a social problem and entails serious health risks. One of the alternatives in the treatment of obesity is the traditional use of medicinal plants, which supports the research and development of obesity phytotherapy. In this article, we provide information about ethnopharmacological species used to treat obesity, through an electronic search of the periodical databases Web of Science, Scopus, PubMed and Scielo, considering the period 1996-2015 and using the descriptors "plants for obesity", "ethnopharmacology for obesity" and "anti-obesity plants" in both Portuguese and English. We analyzed and organized data on 76 plant species, cataloged per the taxonomy, geographic distribution, botanical aspects, popular use, and chemical and biological studies of the listed plants. The anti-obesity effect of the cataloged species was reported, describing actions on the delay of fat absorption, suppression of enzymatic activities, mediation of lipid levels and increase of lipolytic effects, attributed mainly to phenolic compounds. Given these findings, ethnopharmacological approaches are relevant scientific tools in the selection of plant species for studies that demonstrate anti-obesity action. Deeper botanical, chemical, pre-clinical and clinical studies are particularly necessary for species that present phenolic compounds in their chemical structure.
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Maternal obesity is known to predispose offspring to metabolic and neurodevelopmental abnormalities. While the mechanisms underlying these phenomena are unclear, high fat diets dramatically alter intestinal microbiota, and gut microbiota can impact physiological function. To determine if maternal diet-induced gut dysbiosis can disrupt offspring neurobehavioral function, we transplanted high fat diet- (HFD) or control low fat diet-associated (CD) gut microbiota to conventionally-housed female mice. Recipient mice were then bred and the behavioral phenotype of male and female offspring was tracked. While maternal behavior was unaffected, neonatal offspring from HFD dams vocalized less upon maternal separation than pups from CD dams. Furthermore, weaned male offspring from HFD dams had significant and selective disruptions in exploratory, cognitive, and stereotypical/compulsive behavior compared to male offspring from CD dams; while female offspring from HFD dams had increases in body weight and adiposity. 16S metagenomic analyses confirmed establishment of divergent microbiota in CD and HFD dams, with alterations in diversity and taxonomic distribution throughout pregnancy and lactation. Likewise, significant alterations in gut microbial diversity and distribution were noted in offspring from HFD dams compared to CD dams, and in males compared to females. Regression analyses of behavioral performance against differentially represented taxa suggest that decreased representation of specific members of the Firmicutes phylum predict behavioral decline in male offspring. Collectively, these data establish that high fat diet-induced maternal dysbiosis is sufficient to disrupt behavioral function in murine offspring in a sex-specific manner. Thus these data reinforce the essential link between maternal diet and neurologic programming in offspring and suggest that intestinal dysbiosis could link unhealthy modern diets to the increased prevalence of neurodevelopmental and childhood disorders.
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Plant-derived natural products have long-standing utility towards treating degenerative diseases. It is estimated that about two-third of world population depend on traditional medicine for primary medical needs. Fenugreek (Trigonella foenum-graecum Linn.), a short-living annual medicinal plant belonging to Fabaceae family, is used extensively in various parts of the world as herb, food, spice and traditional medicine. Fenugreek is considered as one of the oldest medicinal plants and its health-promoting effects have been cited in Ayurveda and traditional Chinese medicine. The investigations into the chemical composition and pharmacological actions have seen a renaissance in recent years. Extensive preclinical and clinical research have outlined the pharmaceutical uses of fenugreek as antidiabetic, antihyperlipidemic, antiobesity, anticancer, anti-inflammatory, antioxidant, antifungal, antibacterial, galactogogue and for miscellaneous pharmacological effects, including improving women's health. The pharmacological actions of fenugreek are attributed to diverse array of phytoconstituents. The phytochemical analysis reveals the presence of steroids, alkaloids, saponins, polyphenols, flavonoids, lipids, carbohydrates, amino acids and hydrocarbons. This review aims to summarize and critically analyze the current available literature to understand the potential of fenugreek for disease prevention and health improvement with special emphasis on cellular and molecular mechanisms. Current challenges and new directions of research on fenugreek are also discussed. This article is protected by copyright. All rights reserved
Gut microbiota based metabolism of choline produces trimethylamine (TMA) which is further converted to a pro-atherosclerotic metabolite, trimethylamine-N-oxide (TMAO) by flavin monooxygenase (FMO3). Trigonelline from the plant Trigonella foenum-graecum has been reported for the treatment of CVD. Aim of the present study was to check the effect of trigonelline on the gut microbiota based conversion of TMA to TMAO. Trigonelline was isolated from hydroalcoholic extract of seeds of Trigonella foenum-graecum. The isolated trigonelline was characterized through TLC and UPLC-MS. Anaerobic microbe responsible for the metabolism of choline to TMA was isolated by culturing the human gut microbiota in choline enriched medium. The isolated bacteria was identified at molecular level based on PCR amplification of 1500bp of 16S rRNA gene sequence. Isolated FMO3 was used for ex vivo conversion of TMA to TMAO. Further, we investigated the effect of trigonelline in isolated gut microbe based metabolism of choline, lipid profile and TMAO levels in mice with or without suppression of gut microbiota with antibiotics. Liquid-liquid purification and chromatographic analysis confirmed the trigonelline purity (87.26%) and which was also confirmed by mass spectroscopy with m/z 137.4 in positive ionization mode. A total of 30 anaerobic microbes responsible for TMA production were isolated and Citrobacter freundii was the superior among others for the production of TMA. In vitro culture of C. freundii in choline enriched medium supplemented with trigonelline resulted in significantly reduction TMA and followed by TMAO production. In ex vivo, a maximum of 85.3% TMAO production was reduced by trigonelline at concentration of about 300 μg/mL. Serum level of lipids and TMAO were significantly altered in choline fed animals with or without suppression of gut microbiota and this phenomenon was reversed upon the oral administration of trigonelline in a dose-dependent manner. This study demonstrates the effect of trigonelline on gut microbiota responsible for choline metabolism and this can be used as a model for evaluation of herbal drugs and its effect in gut microbiota prompted cardiovascular disorders.
Objective: Galactomannans derived from fenugreek confer known health benefits; however, there is little information regarding health benefits of citrus pectin (CP) and its association with gut microbiome metabolites. The aim of this study was to examine links between galactomannan and CP consumption, microbiota development, and glucose metabolism. Design: Male C57 BL/6 J mice ages 7 to 8 wk were fed ad libitum with a normal diet or one supplemented with 15% of either galactomannan or CP. At 3 wk, an oral glucose tolerance test was performed. Animals were sacrificed at 4 wk and relevant organs were harvested. Results: Fiber enrichment led to reductions in weight gain, fasting glucose levels, and total serum cholesterol (P < 0.05). Compared with mice fed the normal diet, microbiota populations were altered in both fiber groups and were found to be richer in Bacteroidetes rather than Firmicutes (P < 0.05). The modification was significantly greater in galactomannan-fed than in CP-fed mice (P < 0.0001). Also, enhanced levels of the short-chain fatty acid (SCFA) propionate were found in the cecal contents of CP-fed animals (P < 0.05). Protein expression levels of monocarboxylate transporter 1, which may promote transport of SCFA, were measured in the large intestines after fiber consumption. Enhanced adenosine monophosphate-activated protein kinase (AMPK) activation was observed in livers of galactomannan-fed mice (P < 0.05). Conclusion: Consumption of diets containing soluble fibers, as used in this study, resulted in gut microbiota comprising a healthier flora, and led to positive effects on weight, glycemic control, and liver β oxidation via AMPK.