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Observation
Therapeutic potential of the genus Bacteroides in hypertension: an intervention of long-
term intermittent fasting

Junhong Su1, Fanglin Li2, Wenlian Bai3*, Zhongren Ma2*, Maikel P. Peppelenbosch1*

1. Department of Gastroenterology and Hepatology, Erasmus MC – University Medical

Center Rotterdam, Rotterdam, The Netherlands.
2. Engineering Research Center for Key Technology and Industrialization of Cell-based
vaccine, Ministry of Education, Biomedical Research Center, Northwest Minzu University,
Lanzhou, China.
3. Department of Cardiology, People’s Hospital of Linxia Hui Autonomous Prefecture, Linxia,
China.

Corresponding authors:

Maikel P. Peppelenbosch, MD, PhD

Dr. Molenwaterplein 40, 3015 GD, Rotterdam, The Netherlands
Email: [email protected]
Tel: +31107035448

Zhongren Ma, PhD

No. 1 Xibeixincun, 730030, Lanzhou, China
Email: [email protected]
Tel: +86 9312938310

Wenlian Bai, MD
No. 110 Binnan South Road, Linxia, China
Email: [email protected]

Running title: Bacteroides and hypertension

Word count: 1011 (Max:1200; excluding figure legend and references).

NOTE: This preprint reports new research that has not been certified by peer review and should not be used to guide clinical practice.
medRxiv preprint doi: https://doi.org/10.1101/2025.04.10.25325255; this version posted April 14, 2025. The copyright holder has placed this
preprint (which was not certified by peer review) in the Public Domain. It is no longer restricted by copyright. Anyone can legally share,
reuse, remix, or adapt this material for any purpose without crediting the original authors.

Observation
Abstract
Intermittent fasting has shown promise in the management of hypertension, but the
mechanistic explanation for this effect remains largely obscure. Studies in experimental
animals suggest that intermittent fasting acts on hypertension by modifying the microbiome
and particularly by increasing levels of intestinal Bacteroides. Human data, however, are
lacking. Here we conducted a clinical trial [ChiCTR2000034646] to investigate the effects of
15-week intermittent fasting on individuals with hypertension. We observe that long-term
intermittent fasting effectively counteracts high blood pressure, as demonstrated by
significant reduction in systolic blood pressure (144±4.8 [S.E.M.] mmHg at baseline vs.
129±5.6 mmHg at 15 weeks, p=0.004) and diastolic blood pressure (94±5.2 mmHg vs. 79±2.9
mmHg, p=0.005), as well as serum uric acid (410 ± 38 mmol/L at baseline vs. 307 ± 5.5 mmol/L
at 15 weeks, p=0.032), which is a strong risk marker for hypertension. Importantly, this effect
is associated with significant remodeling of the fecal microbiome (p=0.041). Mirroring earlier
data in experimental rodents, we observe a strong inverse correlation between levels of genus
Bacteroides and blood pressure (R=-0.608, p=0.04). Our results strongly support the notion
that the genus Bacteroides is a major determinant of blood pressure in hypertensive
individuals.

Importance: While intermittent fasting is generally recognized to be beneficial for patients

with high blood pressure, the mechanistic basis for this effect is not resolved. Based on
animal data, however, a role of the microbiome and especially the genus Bacteroides has
been suggested. Thus prompted, we conducted a clinical trial in hypertensive individuals to
link the fecal microbiome to changes in blood pressure. We observed strong correlations
between improved blood pressure and fecal levels of the genus Bacteroides. In conjunction
with the body of contemporary biomedical literature our data suggest that the effect of
intermittent fasting on blood pressure is mediated through Bacteroides opening a novel
avenue for rational treatment of this condition.

Key words: Intermittent fasting; Cardiometabolic risks; 16S rRNA gene sequencing; The gut
microbiome; Bacteroides
medRxiv preprint doi: https://doi.org/10.1101/2025.04.10.25325255; this version posted April 14, 2025. The copyright holder has placed this
preprint (which was not certified by peer review) in the Public Domain. It is no longer restricted by copyright. Anyone can legally share,
reuse, remix, or adapt this material for any purpose without crediting the original authors.

Observation
OBSERVATION
Hypertension is a significant public health issue worldwide, responsible for approximately
54.5% of ischemic heart disease and 50.0-58.3% of stroke deaths each year. Poor lifestyle
choices, including obesity and an unhealthy diet, are driving factors for hypertension1.
Although it is well established that the pathogenesis of hypertension is primarily characterized
by decreased vasodilation and increased blood volume, recent studies indicate that dysbiosis
of the gut microbiome is potentially an important factor in its pathogenesis, potentially driving
changes in vasotension2. In this context, it is important to note that drugs used to manage
hypertension can also affect the microbiome, aggravating the dysbiosis and thus
counteracting efficacy3, 4. Identifying the elements in the microbiome that interact with
hypertensive disease would, therefore, be valuable for refining strategies to manage
hypertensive patients effectively.

More specifically, the depletion of short-chain fatty acid (SCFA)-producing bacteria is a critical
feature of a dysbiotic gut microbiota in hypertension2. Notably, Bacteroides, which is a major
producer of intestinal propionate and a significant contributor to the enterotypes in healthy
individuals, is substantially reduced in hypertension5-7. Building on this, research
demonstrates that enhancing the levels of intestinal Bacteroides in animals can help reduce
blood pressure through mechanisms involving the regulation of intestinal steroid hormone
levels5 and the restoration of bile acid metabolism, suggesting that enhancing the levels of
Bacteroides may hold promise in addressing clinical hypertension. However, to date, no
clinical trials have reported associations between altered levels of Bacteroides and the clinical
course of hypertension. This lack of data substantially impedes progress in the field.

Our previous work demonstrated that one month of 16-hour intermittent fasting significantly
increases the level of intestinal SCFAs-producing bacteria in healthy volunteers8. Hence, we
hypothesized that intermittent fasting may improve blood pressure by increasing intestinal
SCFA producers. To directly investigate this notion, we designed a study in which calorie intake
was restricted from 8:00 am to 14:00 pm continuously for 15 weeks (15wksIF) in hypertensive
individuals (Figure 1A). Patients were only allowed to consume no more than 3 bananas or 3
apples or the combination when they felt very hungry before going to bed. Patient baseline
information is shown in Supplementary Table 1.

At the end of 15 wksIF, participants showed substantial improvement in both systolic blood
pressure (SBP) (144±4.8 [S.E.M.] mmHg at baseline vs. 129±5.6 mmHg at 15 weeks, p=0.004)
and diastolic blood pressure (DBP) (94±5.2 mmHg vs. 79±2.9 mmHg, p=0.005) (Figure 1B).
Improvements in clinical hypertension were further objectified by a significant reduction in
circulating uric acid (410 ± 38 mmol/L at baseline vs. 307 ± 5.5 mmol/L at 15 weeks, p = 0.032;
Supplementary Figure 1B), a marker for hypertension9, and indirectly validated by
improvements in two independent risk factors: BMI and liver function (Figure 1C and 1D).
However, changes in blood pressure and associated clinical parameters were not significant
after 4 weeks of fasting, indicating that prolonged fasting is a necessary determinant for the
clinical success of intermittent fasting in lowering blood pressure.
medRxiv preprint doi: https://doi.org/10.1101/2025.04.10.25325255; this version posted April 14, 2025. The copyright holder has placed this
preprint (which was not certified by peer review) in the Public Domain. It is no longer restricted by copyright. Anyone can legally share,
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Observation

To investigate the notion that changes in the gut microbiome-especially with regard to the
levels of the genus Bacteroides-might be instrumental for mediating the effects of fasting on
blood pressure10, the gut microbiome was examined before fasting (week-8) and during
fasting (week 0, 4,9,15) using fecal sampling and 16S sequencing (details in Supplementary
Materials). Employing the data obtained, compositional analysis using Bray-Curtis distance-
based principal coordinate analysis revealed significant changes in the intestinal microbiome
of hypertensive participants after 15wksIF (p = 0.041 by PERMANOVA test) (Figure 1C),
demonstrating substantial shifts in the levels of specific high-abundance taxa.

More specifically, Sankey plot showed that the Firmicutes (71.6 ± 5.92 [S.E.M.] %) and
Proteobacteria (11.93 ± 3.83%), which were the dominating phyla at baseline, decreased to
46.67 ± 11.39% (p=0.0122) and 1.74 + 0.64% (p=0.0401), respectively, following 15wksIF
(Supplementary Figure 1A). In contrast, Bacteroidetes increased from 5.58 ± 1.58% to 44.66
± 11.27 % (p=0.0241), becoming the most abundant phylum. This increase in Bacteroidetes
could be further attributed to a significant rise in sequences from the genus Bacteroides
(Figure 1E), which were 8.01 times higher than before fasting and included primarily B.
stercoris, B. uniformis and B. vulgatus). The change of this genus was validated by Lefse
analysis (Figure 1F) and hierarchical heat tree analysis (Supplementary Figure 1B). Noticeably,
Bacteroides levels were inversely correlated with systolic blood pressure (R=-0.608, P=0.04),
suggesting a causal connection between this bacterium and the control of blood pressure
(Supplementary Figure 1C), supporting the findings in animals by others that changes in
Bacteroides abundance relate to hypertension5.

Interestingly, the administration of living Bacteroides to experimental animals was found to

reduce intestinal endotoxin levels by decreasing the size of E. Shigella compartment11, which
aligns well with the notion that the reduction in E. Shigella in our patients reflects increased
size of the Bacteroides compartment, in turn suppressing Escherichia (Supplementary Figure
1D; Spearman’s ρ=-0.517, p=0.0195). Additionally, intake of rice and wheat-based food during
fasting might also influence microbiome regulation (Supplementary Figure 1E), which
warrants further investigation.

To rule out the possibility that patients who maintained intermittent fasting throughout the
entire study period might have a different baseline microbiome, which could obscure the
effects of fasting, we compared the gut microbiome between non-completers and completers
at baseline (0 weeks). No significant differences were observed in microbiome diversity
(P=0.4209) (Supplementary Figure 1F), microbiome composition (P=0.304 by ANOSIM test)
(Supplementary Figure 1G) or the relative abundance of the Bacteroides (P=0.2622)
(Supplementary Figure 1H). Hence the most straightforward explanation is that a strong
negative correlation exists between blood pressure and levels of intestinal Bacteroides.

Conclusion
We observed that 15wksIF is effective in improving hypertension by increasing
medRxiv preprint doi: https://doi.org/10.1101/2025.04.10.25325255; this version posted April 14, 2025. The copyright holder has placed this
preprint (which was not certified by peer review) in the Public Domain. It is no longer restricted by copyright. Anyone can legally share,
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Observation
intestinal Bacteroides levels, although patient’s motivation and capacity to maintain such a
regimen may be limited, particularly in males12. Furthermore, this study provides a promising
perspective that Bacteroides may serve as a novel bacterial candidate for restoring an
imbalanced gut microbiome and improving blood pressure. This insight opens new avenues
for the rational design of beneficial Bacteroides-based therapeutic strategies to address this
condition, with potential to complement current treatment that carry side effect and
exacerbate dysbiosis of the gut microbiome.

Competing interests
The authors declare that they have no competing interests.

Acknowledgements
The authors would like to acknowledge Yueying Wang and Yuxin Su for their assistance in
sample collection and questionary survey. The authors are grateful to all patients who
contributed to this work. This study was supported in part by grant (KICH2.V4P.22.015) of the
Dutch Organization for Scientific Research and the Dutch Cancer Foundation.

Data availability
The sequence data have been deposited with links to BioProject accession No.
PRJNA1118570 in the DNA Data Bank of BioProject database
(https://www.ncbi.nlm.nih.gov/bioproject/PRJNA1118570). Source data of figures is
included in the supporting files.

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Observation
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preprint (which was not certified by peer review) in the Public Domain. It is no longer restricted by copyright. Anyone can legally share,
reuse, remix, or adapt this material for any purpose without crediting the original authors.

Observation
Figure 1. Effects of intermittent fasting on blood pressure and the gut microbiome in
hypertensive patients.

(A) Study flowchart. The change in blood pressure (B), an independent risk factor BMI (C)
and liver function indicator ALT (D). Data are expressed as mean ± S.E.M. P values are
calculated using paired student’s t-test (one-tailed). (E) The overall microbiome composition
in patients was represented visually using principal coordinate analysis (PCoA) of genus-level
relative abundance. The samples that are clustered as individual centroids are from different
time point denoted by different colors. P values were calculated using PERMANOVA test. (F)
Sankey plots showing the longitudinal change of genus level abundance during intermittent
fasting. P values are calculated using paired student’s t-test (one-tailed). (G) Taxa that
differed at baseline at week 0 versus end of fasting at week 15 are depicted and visualized in
bars using LEfSe with a log LDA (linear discriminant analysis) score above 4.0. Blue bar
represents taxa where increases in its relative abundance or prevalence associated with
15wksIF, while red bar means taxa decreased during 15wksIF.