Probiotics and mood enhancement -
A fermented milk drink kefir that contained Lactobacillus casei strain Shirota prevented stress-related cortisol increases and raised serotonin levels in stressed medical students.
Lactobacillus helveticus NS8 was compared to the SSRI, citalopram, in rats with depression, anxiety, and cognitive dysfunction due to chronic stress.
It lowered cortisol and restored serotonin and other brain neurochemical levels to normal. Other Lactobacillus helveticus-containing probiotics have also been shown to reduce stress-related depression and anxiety by affecting serotonin, cortisol, and other neuroactive compounds in animal studies.
Prebiotics, like probiotics, can also act as important regulators of mood and brain function. In a recent study, prebiotics were found to decrease the secretion of the stress hormone, cortisol, and improve emotional processing in healthy volunteers.
Participants received one of two prebiotics fructooligosaccharides, FOS, or Bimuno-galactooligosaccharides, B-GOS or a placebo maltodextrin daily for three weeks. Morning cortisol levels were significantly lower after B-GOS intake compared with placebo.
Participants taking B-GOS also showed increases in the processing of positive versus negative attentional vigilance, which is an indication that the prebiotic had anti-anxiety effects.
No effects were found after the administration of FOS. Overall, the results of these studies show that psychobiotics have the potential to positively impact brain function, boost mood, treat depression and anxiety, and help you handle stress.
The best psychobiotics and the best dosages for those psychobiotics have yet to be determined, but a number of them used in the studies described above are commercially available in probiotic tablets. Good luck, and may your gut-brain connection be healthy! pii: S 15 As a service to our readers, University Health News offers a vast archive of free digital content.
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The questionnaire measures 10 bowel habits, asking whether there were improvements in the following parameters in the last 4 weeks: overall bowel health; frequency of defecation; amount of defecation; feeling of incomplete evacuation; stool odor; abdominal cramping; bowel sounds; number of gas passage; abdominal distention; and frequency of diarrhea.
Extracted genomic DNA was confirmed via gel electrophoresis and was quantified by spectrophotometer NanoDrop ND Thermo Scientific, Waltham, MA. Hypervariable regions V3-V4 of 16S ribosomal ribonucleic acid rRNA gene were amplified using barcoded universal primers for each sample.
Polymerase chain reaction PCR was carried out by using BioFact F-Star taq DNA polymerase BioFACT, Seoul, Republic of Korea. PCR reactions were amplified using a GeneAmp PCR system Applied Biosystems, Foster City, CA.
The PCR program was as follows: initial for 5 minutes hold at 94°C, followed by 28 cycles of denaturation 30 seconds, 95°C , annealing 30 seconds, 60°C , and extension 30 seconds, 72°C , with a final extension step 10 minutes, 72°C followed by holding at 4°C.
The amplified products were purified with PureLink Quick Gel Extraction and PCR Purification Combo Kit Invitrogen, Carlsbad, CA and quantified by the Qubit 2. The size of library was assessed by BioAnalyzer Agilent Technologies, Santa Clara, CA.
The amplicons were pooled and sequenced with an Illumina MiSeq sequencing system Illumina, San Diego, CA. Microbial sequences were processed using QIIME2 version Briefly, sequences were denoised to remove the sequences with low-quality score and chimeras via DADA2.
A phylogenetic tree was generated using MAFFT and FastTree method for diversity analyses. Downstream analyses on alpha diversity were carried out to measure dissimilarities in richness and evenness of microbial community.
Comparisons of relative abundance between groups were performed to identify the differential features across the samples. A validated item self-reported questionnaire was used to ask the level of stress in a category of burn-out, depression, and anger during the past 1 month Total scores were calculated, and higher scores mean higher level of stress.
The quality of life QoL was measured with the Satisfaction With Life Scale SWLS , a validated subjected report of global life satisfaction Responses were summed and higher scores indicate higher QoL. The Korean version of Geriatric Depression Scale GDS-K was used to evaluate the level of depression.
The GDS-K is a item self-reported questionnaire which is a validated instrument for the diagnosis of clinical depression The cumulative score is calculated, and the higher score means the higher level of depression.
The Positive Affect and Negative Affect Schedule PANAS is a validated self-report instrument in the assessment of positive and negative affect The PANAS is comprised of two item scales which measure both positive and negative affect, respectively.
The summed scores from each positive and negative affect indicate the level positive and negative affect, respectively. Briefly, sample or standard was added to a plate coated with capture antibody and the plate was incubated for 2 hours at room temperature. After washing the plate sufficiently, detection antibody was added to the plate and the plate was incubated for 2 hours at room temperature.
Streptavidin conjugated to horseradish peroxidase was added to each well and the plate was incubated for 20 minutes at room temperature.
Then substrate solution was added to each well for 20 minutes of incubation at room temperature following sufficient washing with wash buffer, and the plate was ready for determining the optical density at nm wavelength using a microplate reader SpectraMax iD3, Molecular Devices, Austria.
The normality assumption and homogeneity of variance were tested by Kolmogrov—Smrinov test for study variables. To compare the difference between groups at each time point and delta value between the visits Δ Week 4—Week 0 , Δ Week 8—Week 0 , and Δ Week 12—Week 0 between the two groups, we performed unpaired t -test, Mann—Whitney U test or generalized linear model GLM.
To compare the difference between baseline and the data from end point Week 12 , we used paired t -test or Wilcoxon signed rank test. To detect the difference between groups over the visits, we used a mixed-model analysis of variance ANOVA or Friedman test as a nonparametric alternative to the repeated measures ANOVA.
Correlations were assessed by Spearman rank correlation analysis. Correction for multiple testing was performed based on the false discovery rate or Bonferroni correction.
All statistical analyses were conducted using Partek version 6. A total of volunteers were screened for eligibility and 63 subjects were enrolled for the study Supplementary Figure 1.
Sixty-three participants were randomized, with 31 and 32 subjects in the placebo and probiotics group, respectively. Fifty-three individuals completed the study and 10 participants withdrew the consent and discontinued the study, and no clinically relevant adverse events were reported during the intervention.
Therefore, we confirmed that randomization was successful. In all analyses, we included data from participants who completed the intervention. Demographic and clinical characteristics at baseline are summarized in Supplementary Table 2.
Average age was The ratio of male to female and BMI did not significantly differ between two groups. Socioeconomic characteristics, including educational level, marital status and type of household, and other health-related characteristics such as cigarette use, alcohol use, physical activity, and self-evaluated health status, were not different between the placebo and probiotics group.
In addition, cognitive functions and depression scores, determined by MMSE and Geriatric Depression Scale GDS-K , were not different between two groups at the baseline.
To assess the effect of probiotics on intestinal health, participants filled questionnaire at 4th week, 8th week, and 12th week, respectively, asking whether there were improvements in bowel habits in the last 4 weeks. Bowel habits such as frequency and amount of defecation; feeling of incomplete evacuation; stool odor; number of gas passage; bowel sounds; and abdominal distention were not significantly improved both in the placebo and probiotics group during the intervention period data not shown ; however, scores in frequency of gas passage and abdominal distention showed significant improvements in the probiotics group compared with the placebo 3.
Beneficial influence of probiotic supplementation on intestinal health and gut microbiota. A,B Improvement scores in frequency of gas passage and abdominal distention measured at each visit are shown.
Data are presented as mean SEM. C Relative abundance of the gut microbiota at the phylum level and D—G at the genus level was measured throughout the intervention.
Data are presented as mean min-max. In order to address whether the improvement was driven by any changes in the intestinal bacterial communities, gut microbiome profiling analysis was performed in all participants. Bacterial genomic DNAs from stool samples collected at baseline, 4th week, 8th week, and 12th week were sequenced using 16S rRNA sequencing technology.
After preprocessing of bacterial sequences for quality control as described in the Methods, we obtained a total of 10,, raw reads and average reads of 80, per sample. We found no significant changes in the diversity both in the placebo and probiotics group during the intervention period Supplementary Figure 2A—D.
Further, we compared the relative abundance of OTUs and specific bacterial taxa at the different phylogenetic levels. Microbial composition at OTU level showed no significant differences during the intervention both in the placebo and probiotics group Supplementary Figure 2E.
At the phylum level, no significant changes in relative abundance were detected during the intervention both in the placebo and probiotics group Figure 1C.
However, at the genus level, we found significant changes in the gut microbial composition in the probiotics group and no changes in the control group Figure 1D—G. The assessment was performed at baseline, Week 4, and Week 12 Table 1. The changes at the fourth week from baseline in the probiotics group were not different from those in the placebo group for all the domains of the cognitive assessment; however, the changes at Week 12 from baseline in the scores of mental flexibility test were significantly different between placebo and probiotics group Table 2.
In addition, study subjects filled series of questionnaires to evaluate the impact of probiotics on mood status including quality of life, stress, depression, and positive and negative affect. The week consumption of probiotics did not change the scores of quality of life, GDS-K, and PANAS; however, it did affect the stress score Table 2.
While the stress score was increased in the placebo group 1. Note : Data are presented as mean SD. Improved cognitive and mental functioning after probiotic supplementation.
A Change from baseline of cognitive performance score in the mental flexibility test is shown. Reduction in the performance score of mental flexibility indicates improved attention and executive function. B Change from baseline of stress level is shown.
Reduction in the change indicates a reduced level of mental stress. The observations that probiotic supplementation improved the cognitive function and mental stress prompted us to determine the level of BDNF in blood. BDNF is a neurotrophic factor known to be crucial for learning, memory function, and stress.
In addition, to address the question of whether changes in intestinal bacterial communities be related to the serum level of BDNF, we conducted correlation analysis between the relative abundance of each genera and the level of BDNF.
These findings suggest that reduction in the relative abundances of Eubacterium and Clostridiales in the gut driven by probiotic supplementation closely related to the increase in the serum BDNF, thereby improving brain functions. Elevated level of serum BDNF after probiotic supplementation.
A Change from baseline of serum BDNF is shown. B Scatter diagrams with regression lines show the relationship between relative abundance of shifted gut bacteria after probiotic supplementation and the level of serum BDNF.
Measurements were rank-normalized and plotted separately for the placebo and probiotics group. Correlation coefficient R S and p -values based on Spearman rank correlation analysis.
In the present study, we conducted a randomized, double-blind, placebo-controlled, multicenter trial to address the impact of probiotics on intestinal health and how they contribute to ameliorating cognitive and mental decline in the older adults.
Our findings demonstrate that probiotics have system-wide effects on the gut—brain axis in healthy community-dwelling older adults by promoting cognitive and mental health and changing the gut microbial composition.
However, while neurodegenerative disorders and psychological distress are a common threat to well-being in old age, nutritional intervention to prevent or delay age-associated decline in brain function in the general older population is still underexplored.
In fact, there is only one report on the effects of probiotic consumption in healthy older adults, showing that milk fermented by Lactobacillus helveticus IDCC improved cognitive functions in healthy older adults However, the sample size was too small and the criteria for the study participants did not represent the general population of older adults.
Therefore, the critical need for clinical studies in the general population has been raised. To the best of our knowledge, this is the first well-controlled clinical study demonstrating system-wide effects of probiotics on the gut—brain axis, which encompasses the large-scale analysis of the gut microbiota and multiple aspects of brain functions in healthy older population.
Randomized controlled trials RCTs are very challenging for several issues but the most rigorous method, which provides the most reliable evidence for clinical practice; however, there are few RCTs specifically designed for older adults because it is difficult to recruit older people, particularly community-dwelling older adults 34— Therefore, older adults have been excluded from clinical trials and most studies focused on older group of patients In addition, it is relatively hard to follow up and contact older people during a trial which increases the risk of dropout and reduces compliance 36 , Despite these challenges, in the present study, participant compliance was good as the average rate of compliance to intervention was Moreover, it is important to note that the present study recruited older adults without diseases, not focusing on good responders to a treatment effect such as patients with neurological disorders, which makes our findings more applicable as a generalized health care strategy in community-dwelling older population.
In the probiotics group, the gut microbial composition shifted gradually, and the most relevant change was the reduction in the abundance of bacteria that cause inflammation including Eubacterium , Allisonella , and Prevotellaceae. It has been identified that Eubacterium and Prevotellaceae species, which were significantly reduced after probiotic consumption, are proinflammatory microbiota associated with autoimmune disease and chronic intestinal inflammation in mice 38 , Of note, the genus Allisonella , whose abundance was significantly reduced in the probiotics group, produces histamine, a biogenic monoamine inducing proinflammatory response both centrally and systemically Therefore, these findings may parallel our hypothesis that probiotic supplementation in the older adults may negatively affect inflammaging, a characteristic of chronic low-grade inflammatory status in older adults, via the modulation of microbial composition.
However, further studies are required to assess whether the probiotic supplementation affects immunological mechanisms. The findings of the present study suggest that interaction between the gut microbiota and the central nervous system may underlie the improvements in cognitive and cerebral functioning upon probiotic supplementation and explain the concomitant changes in peripheral neuromodulators.
BDNF, a neurotrophic factor vital for synaptic formation, plasticity, and neuroimmune responses, has long been studied to assess its critical role in learning, memory formation, and affective disorders 41 , Previously, the influence of diet and nutrition on BDNF has been explored; and serum BDNF has been shown to be increased in response to dietary supplements in humans.
For example, a 1-week of oral consumption of α-linolenic acid increased the level of serum BDNF in healthy young adults Also, a 6-week supplementation with natural extracts rich in flavonoids and polyphenolic compounds enhanced serum BDNF levels in physically active men In the present study, it was notable that the beneficial impact of a week probiotic intervention on serum BDNF levels was evident in older population.
More recently, BDNF has emerged as a pivotal link in the gut—brain axis 41 , Several studies demonstrated that gut dysbiosis correlates with reduced expression of BDNF, which alters cognitive function and triggers anxiety-like behavior in germ-free animals 45 , 46 , supporting a role of BDNF in the gut—brain axis.
Interestingly, we observed that the relative abundance of significantly shifted gut microbes correlated with the level of serum BDNF in the probiotics group only. This indicates that administration of probiotics may affect the interaction between the gut microbiome and the host BDNF, thereby improving brain functions.
Overall, the evidence from this study shows that the shifts in microbial community mirrored changes in the cognitive and mental scores. Several mechanisms could explain the interaction between changes in abundance of commensal bacteria and brain function observed in the probiotics group.
First, it is plausible that the production of neurotransmitters, such as γ-Aminobutyric acid GABA , dopamine, acetylcholine, serotonin, by commensal bacteria, and neurochemicals including BDNF, may directly or indirectly modulate cognition and mood status As shown in a previous study, probiotic administration influences GABA receptor throughout the brain, with reduced stress-induced anxiety- and depression-like behaviors in rodents Moreover, inflammation-mediated pathways might initiate the pathogenesis of neurodegeneration via the microbiota—gut—brain axis.
With respect to inflammaging during normal aging, chronic low-grade inflammation in older adults may affect neuroinflammation by modulating glial cells, which stimulates cognitive impairment One of the routes to translate systemic inflammatory signals into the brain is stimulation of microglia by peripheral cytokines that cross the blood—brain barrier, leading to a proinflammatory status in the brain and dysregulation of neurological processes Additionally, the immunomodulatory roles of circulating immune cells in neuroplasticity also affect the expression of BDNF 49 , Therefore, it is plausible that mitigation of inflammaging in older adults with probiotic intervention might positively impact on cognitive and mental functions via the modulation of BDNF signaling.
Further studies are needed to clearly demonstrate the effect of probiotics on inflammatory status and gut microbiome at the functional level.
The present study is not without limitations. First, direct evidence of improvement in peripheral and cerebral inflammation by probiotic consumption is lacking, which might be the crucial interface linking the gut—brain axis in the present study.
Therefore, further mechanistic studies might be needed to elucidate the role of probiotic supplementation by finding biomarkers to link the axis.
Additionally, although our results indicated that the benefit of 3 months duration of probiotic intervention was evident, there were no significant changes in some of the cognitive functions in the neuropsychological assessment battery in which we assume that the study duration was not enough to monitor the improvements.
Therefore, further studies are required with a longer period of intervention. Despite these limitations, this is the first study examining the effects of probiotic supplementation on brain functions in community-dwelling older adults. In conclusion, our study showed that probiotic supplementation is beneficial for improving cognitive and mental health in community-dwelling healthy older adults with changes in gut microbial composition.
These results provide evidence that probiotics have health-promoting properties as part of a healthy diet in the general population of independently living older adults.
designed the research; C. conducted the research; C. performed bioinformatic and statistical analyses and analyzed the data; W. conducted the statistical analysis; and C. and D. wrote the manuscript. The authors thank S. for his assistance in recruitment of participants; D. had primary responsibility for final content.
All authors read and approved the final manuscript. This research was funded by the National Research Foundation of Korea NRFS1A5A2A , the Promoting Regional specialized Industry R , the Ministry of Trade, Industry and Energy MOTIE , and Korea Institute for Advancement of Technology KIAT , Republic of Korea.
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