Skip to main content
Download PDF

The potential of novel gut microbiota supplement in mitigating gut inflammation, alleviating oxidative stress linked to aging, and improving cognitive function in aged mice

BMC Complementary Medicine and Therapies volume 25, Article number: 137 (2025) Cite this article

Abstract

Background

Aging is a physiological process that impacts multiple systems of organs. Alzheimer’s disease (AD) is the most common form of dementia in the elderly, and it is a major problem in aging societies. The development of AD is linked to an accumulation of amyloid beta and Tau proteins, which impair cognition and cause memory loss.

Purpose

We studied whether probiotics strains could protect and how effectively probiotics might delay age-related changes.

Methods

Two probiotics, Lactobacillus paracasei MSMC39-1 and Bifidobacterium animalis MSMC83 strain, were administered orally to mice beginning in middle age and continuing into aged mice. The mice were subsequently monitored and assessed for inflammation and oxidative stress in the colon, brain, and liver tissues, as well as for overall health, over a period of 16 weeks.

Results

We found aged mice received the combination of these probiotics showed a lower level of inflammatory markers and improved overall health compared to the control group. MSMC39-1 and MSMC83 enhance gut integrity and general well-being in aged mice and result in improved cognitive memory.

Conclusion

Our findings suggest that these probiotics supplements may be particularly useful in strategies for the prevention of age-related pathologies by reducing inflammation and oxidative stress, which in turn would slow disease progression.

Clinical trial number

Not applicable.

Peer Review reports

Introduction

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder specifically characterized by slow deterioration in cognition, memory and activities of daily living [1, 2]. For example, in Alzheimer’s disease one of the major factors involved is build-up of abnormal proteins across brain regions resulting in neuron loss along with chronic inflammation within nervous tissue. The accumulation of amyloid-β (Aβ) peptide in the brain causes pathological processes such as neurofibrillary tangles (NFTs), inflammatory responses, increased oxidative stress causing neuronal tissue disruption [3]. Hyperphosphorylation of Tau protein can trigger the activation of inflammatory mediators such as activation of Toll-like receptor (TLR)-4 in human macrophages, resulting in neuroinflammation in tauopathies [4]. Ongoing of aging causes not only brain disruption but also has influence on a metabolism, metabolic syndrome in the elderly is associated with increased obesity, β-cell dysfunction, and sarcopenia, in addition to dementia.

Metabolic dysfunction may lead to the development of insulin resistance, type 2 diabetes, and obesity. Probiotic supplements research has gained attention because of its potential therapeutic implications for various conditions particularly in the setting of aging [5]. As the global population continues to age, there is an increasing focus on maintaining health and well-being in older adults. Probiotics, which are live microorganisms that confer health benefits when consumed in adequate amounts, have emerged as a promising area of research in this context. Moreover, there is growing evidence to suggest that probiotics may have a positive impact on mental health, which is particularly relevant for the aging population. The gut-brain axis, a complex communication network linking the gut and the brain, indicates that gut health can influence mood and cognitive function. The brain and gut communicate through several factors, which all have an impact on intestinal permeability, endocrine signaling, and immune system activation [6]. Evidence suggests that Lactobacillus paracasei holds promise for improving cognitive deficits associated with aging and Alzheimer’s disease, as observed in experimental mouse models. For example, supplementation with L. paracasei PS23 in senescence-accelerated mouse models (SAMP8) has demonstrated the ability to slow the advancement of cognitive decline associated with aging [7, 8]. Anti-senescent effect was achieved by reducing anxiety-like behaviors and memory impairment, likely through the induction of antioxidant and anti-inflammatory actions. Corpuz and colleagues have indicated that prolonged dietary supplementation with L. paracasei K71 was achieved by mitigating age-related cognitive decline by enhancing the expression of BDNF in the hippocampus [9]. Another study has demonstrated that heat-killed L. paracasei D3-5 is effective in both preventing and treating age-related leaky gut and inflammation, leading to enhancements in physical and cognitive functions in both elderly mice and humans [10]. Moreover, research suggests that Bifidobacterium animalis could be beneficial in mitigating the symptoms of Alzheimer’s disease (AD) as observed in mouse models. The combination of B. animalis and Bifidobacterium longum protects older mice from cognitive decline due to D-galactose treatment, alleviating symptoms like anxiety, uncoordinated movement, hippocampal aging, neuroinflammation, and oxidative stress by upregulating Sirt 1 [11]. Collectively, the positive outcomes of this study are probably a result of its effectiveness in modulating inflammation, managing oxidative stress, and enhancing gut-brain communication. In our previous study, we demonstrated that L. paracasei MSMC39-1 has considerable anti-inflammatory capabilities [12]. In studies involving rats with dextran sulfate sodium (DSS)-induced colitis, indicated that this probiotic strain successfully diminished tumor necrosis factor-alpha (TNF-α) secretion in both colon and liver tissues. This suppression was associated with reduced levels of aspartate transaminase (AST) and alanine transaminase (ALT) in the serum, indicating a protective effect against inflammation-induced tissue damage [12]. B. animalis MSMC83 is a probiotic strain studied for its health benefits, particularly against oxidative stress. Our previous research shows that its supplementation significantly enhances antioxidant properties. In rat experiments with D-galactose-induced oxidative stress, this probiotic increased the activity of key antioxidant enzymes like superoxide dismutase, catalase, and glutathione peroxidase, while reducing malondialdehyde levels, a marker of oxidative stress [13]. Therefore, we speculated that the aging-related phenotypic features might be improved by incorporating these probiotic strains that efficiently lower inflammation and oxidative stress. We here show these probiotics boosted liver and brain health by reducing inflammation, oxidative stress, and cognitive decline in our study on elderly mice.

Materials and methods

Probiotic strains and culture condition

Probiotic strains, Lactobacillus paracasei MSMC39-1 and Bifidobacterium animalis MSMC83 were obtained from the Center of Excellence in Probiotics, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand. L. paracasei MSMC39-1 and B. animalis MSMC83 were cultured in de Man-Rogosa-Sharpe (MRS) medium (HiMedia Lab., Mumbai, India) at 37 °C under anaerobic conditions for 24 and 48 h. Subsequently, the density of bacterial suspensions was adjusted to.

10 9 colony forming units (CFU)/ml for the oral administration in mice.

In vivo treatment

Male C56BL/6 mice (Mus musculus) were purchased from Nomura Siam International Co, Ltd (Bangkok, Thailand). All animal procedures were performed according to the guidelines of the Ethics and Research Standardization Section, Srinakharinwirot University (approval number: COA/AE-015-2564). Mice were maintained at room temperature (22 ± 1 °C), 55 ± 10% humidity, and on 12 h light/12 h dark cycle. The study began when the mice reached the age of two months. Mice were divided into four groups for the study: old control (OC) group, young control (YC) group as a control group while middle-aged group (MP) and aged group (AP) mice were supplemented with L. paracasei MSMC39-1 and B. animalis MSMC83. In the AP group received 200 µL of L. paracasei MSMC39-1 and B. animalis MSMC83. oral gavage at 109 CFU/mL starting at 14 months, while the MP group began supplementation at.

10 months of age until 20 months of age. The OC and YC groups were euthanized and subsequently analysed at 20 months and 6 months of age, respectively. Mice were monitored on a daily basis and were euthanized via cardiac puncture under isoflurane anaesthesia. Mice were initially anesthetized in an induction chamber containing 5% isoflurane while their awakening state and respiratory movements were closely monitored. Abdomen of the mice was accessed by incising the diaphragm, followed by the removal of the pericardium. A cardiac puncture was then carried out on the left ventricle, enabling the collection of 1-1.5 ml of blood. After this procedure, the mice peacefully died.

Morris water maze Spatial test

A pool with white color and the diameter of 140 cm and a depth of 40 cm was used in this study and filled with water (25 ± 2 °C). The pool had four quadrants, one of which was a target quadrant with a plexiglass platform placed in it. The platform was invisible and placed 1 cm under water. Mice were trained for five days, and a probe test was conducted in one day. Each training day contained four trials that took 60 s and mice were allowed to rest for 15 s, and then the next trial started. Finally, in probe test (6th day of the test), the platform was removed from the pool, and rats were released randomly to one quadrant and they were allowed to swim for 60 s.

Biochemical analysis

At the end of the experiment, mice were fasted for 12 h prior to euthanasia. Plasma was collected and centrifuged of heparinized blood at 1,500× g, 4 ◦C for 15 min. Biochemical parameters, including total sugar, total cholesterol, triglyceride, high-density lipoprotein (HDL), low-density lipoprotein (LDL), aspartate transaminase (AST), and alanine transaminase (ALT), were measured by the Professional Laboratory Management Corp, Co., Ltd. (Bangkok, Thailand).

Evaluating oxidative stress in mice

Tissue homogenates of the brain, liver, and colon were prepared to assess oxidative stress levels. Oxidative stress markers, including superoxide dismutase (SOD) and malondialdehyde (MDA), were measured following the instruction procedure. The superoxide dismutase kit (706002, Cayman Chemical Company, Ann Arbor, Michigan), and thiobarbituric acid reactive substances (TBARS) assay kit (#10009055, Cayman Chemical Company, Ann Arbor, Michigan) were utilized for this purpose.

Tissue homogenates cytokine measurement by Enzyme-Linked immunosorbent assay

(ELISA)

Kidney tissue was meticulously dissected and briefly washed with chilled 1X PBS to remove any remaining blood. The tissue was then cut into smaller pieces on ice. Following this, the tissue was transferred to a homogenizer along with RIPA buffer that included a protease inhibitor. The tissue sample underwent centrifugation at 10,000 x g for 20 min at a temperature of 4 °C to sediment the cellular debris, after which the supernatant was carefully transferred to a new microfuge tube and stored at -80 °C.The levels of TNF-α and IL-1β in the supernatant of brain, liver, and colon homogenates were quantified using the TNF-α DuoSet® ELISA kit (#DY410-05, R&D systems, Minnepolis, MN) and mouse IL-1 beta/IL-1F2 DuoSet ELISA mouse IL-1 beta/IL-1F2 DuoSet ELISA (#DY401-05, R&D systems, Minnepolis, MN) following the manufacturer instructions. The reaction was detected by adding streptavidin-HRP (#DY999, R&D systems, Minnepolis, MN) and their substrate color reagent A and B (#DY999, R&D systems, Minnepolis, MN). Plate was read at 450 nm using a microplate reader Biochrom (Anthos, 2010).

Histological and immunohistochemistry analysis

Liver, brain (hippocampus) and colon were harvested, rinsed, and weighed after blotting them dry. After that, the liver, hippocampus, and colon were fixed with 4% (v/v) formaldehyde. The brain, liver, and colon tissue were labeled with hematoxylin and eosin (H&E) for further staining. In addition, the liver tissue with Oil Red O and the colonic tissue were labeled with zonula occludens (ZO)-1 (used at 1:100, #617300, ThermoFisher Scientific) and occludin (used at 1:100, #711500, ThermoFisher Scientific), for further immunohistochemical staining. The morphology was observed under a light microscope (Olympus BX53, Olympus Corporation, Tokyo, Japan).

Gene expression analysis by qRT-PCR

Total RNA was extracted using the RNA-spin™ total RNA extraction kit (#17211, iNtRON Biotechnology, Inc) following under the manufacturer’s protocols. RNA concentration was confirmed using NanoDrop™ 2000/2000c Spectrophotometers (#ND-2000, ThermoFisher Scientifi). After that, total RNA was reverse transcribed to cDNA using Maxime RT premix kit (#25131, iNtRON Biotechnology, Inc) following the manufacturer’s protocols. mRNA expression was analyzed by Bio-rad CFX96 real-time system & c1000 (Bio-Rad Laboratories, Inc.) on RealMOD™ Green W² 2x qPCR mix. After that, threshold (Ct) was used for calculating gene expression. The primer sequences used for qRT-PCR were β-actin (internal reference), sense 5’- AGAGGGAAATCGTGCGTGAC-3’ and antisense 5’- CAATAGTGATGACCTGGCCGT-3’, Toll-like Receptor 2 (tlr2), sense 5’- AGACAAAGCGTCAAATCTCAG-3’and antisense 5’-GCATCACATGACAGAGACTCC-3’, Toll-like Receptor 4 (tlr4), sense 5’-TCCCTGCA TAGAGGTAGTTCC-3’ and antisense 5’ CAAGGGGTTGAAGCTCAGATC-3’, Amyloid Beta Precursor protein (app), sense 5’-TGGCCAACATGATTAGTGAACC-3’, and antisense 5’-AAGATGGCATGAGAGCATCGT-3’, Microtubule Associated Protein Tau (tau), sense 5’-CTCAGCATCCCACCTGTAAC-3’, and antisense 5’-CCAGTTGTGTATGTCCACCC-3’.

Statistical analysis

The data were analyzed using GraphPad Prism 9 software. The bars on the graph depict mean ± SD. The statistical analysis of the data was carried out using student t test and one-way analysis of variance (ANOVA). Values of P < 0.05 *, P < 0.01 **, and P < 0.001 *** were considered statistically significant.

Results

The addition of probiotics improved the colonic structure in mice that are aging naturally

Aging is characterized by a decline in cellular function and metabolism. This decline results in heightened oxidative stress and inflammation, both of which are linked to degenerative diseases. We sought to investigate the impact of different durations of probiotic treatment on aged mice, specifically comparing middle-aged and aged mice. We administered a combination of two probiotic strains to middle-aged (10 month old) and aged mice (14 month old), as specified in the experimental protocol (Fig. 1A). Following the administration of probiotics to aged mice, we noted that both the MP and AP groups exhibited a normal physical condition, with no substantial alterations in body weight when compared to the OC group. A histological examination of colon tissues, accompanied by H&E staining, revealed a reduced quantity of crypt structures in the OC group, alongside an increased presence of immune cell infiltration in the OC group relative to the MP and AP groups (Fig. 1B). The length of the crypt was observed to be greater in the OC group when compared to the YC group but was normalized in both the MP and AP groups (Fig. 1C).

Fig. 1
figure 1

Potential of probiotics protects against structural changes in the intestines of elderly mice. AP: aged control group, YC: young control group, MP: middle-aged group and OC: old control group. (A) The schematic diagram illustrates the protocol for treating elderly mice with probiotics, as well as the control group of mice, along with a timeline. (B) Representative of histological colon tissue from elderly mice that were administered probiotics were compared to those from their respective control groups. The images show cross-sectional views of the colon tissue at 4x magnification (upper panel) and 20x magnification (lower panel). (C) The quantitative graph depicting the length of crypts shows discrepancies among four separate groups (number of fields = 5 in each section, number of mice (n = 3 per group) The results shown in this figure are expressed as the mean ± SEM. An one-way ANOVA was used to analyze the comparisons of more than two groups followed by Tukey’s multiple comparison tests. P < 0.05 *, P < 0.01 **, and P < 0.001 ***

Full size image

Elderly mice exhibited inflammation in the gastrointestinal tract, while the addition of probiotics resulted in a reduction of gut inflammation

The intestinal barrier functions as the principal physical safeguard against microbial invasion within the gut, and its structural integrity is essential for preventing inflammation caused by pathogens. Our research indicated that the expression of intestinal barrier proteins, specifically Occludin and ZO-1, did not exhibit notable variations between the AP and MP groups in comparison to the OC group. Nevertheless, the expression levels of these markers were observed to be elevated in the OC group relative to the YC group (Fig. 2A). The mRNA expression levels for tlr2 and tlr4 were markedly increased in OC, but returned to baseline in both MP and AP following the administration of a combination probiotic. In the colons of aged MP and AP mice, the levels of proinflammatory cytokines IL-1β and TNF-α were seen to decrease following probiotic treatment, showing similar concentrations in both groups (Fig. 2B). This implies that probiotic supplementation could effectively reduce gut inflammation in aged mice.

Fig. 2
figure 2

The influence of probiotics on gut barrier and inflammatory-related molecules expression in aged mice. AP: aged control group, YC: young control group, MP: middle-aged group and OC: old control group. (A) In the upper panel, the immunohistochemistry of Occludin expression is depicted, while in the lower panel, the ZO-1 expression in colon tissue of elderly mice that were administered probiotics (from n = 3 sections) (B) Relative mRNA expression levels of inflammatory related molecules in colon tissue. (n = 4 mice in each group) (C) Proinflammatory cytokine expression in the colon tissue of elderly mice that were administered probiotics was compared to that of their respective control groups. (n = 4–5 mice in each group) The results shown in this figure are expressed as the mean ± SEM. An one-way ANOVA was used to analyze the comparisons of more than two groups followed by Tukey’s multiple comparison tests. P < 0.05 *, P < 0.01 **, and P < 0.001 ***

Full size image

The impact of probiotic on lipid metabolism in elderly mice

The rise of sterile inflammation is correlated with advancing age and is linked to the onset of numerous diseases. A crucial element of the aging process includes alterations in lipid metabolism. To investigate the potential benefits of a probiotic combination on lipid metabolism, we assessed lipid metabolic markers, such as total cholesterol and triglyceride concentrations, in the bloodstream of aged mice. We later found that administering probiotics led to lower levels of those markers in aged mice from both the AP and MP groups (Fig. 3A). The levels of cholesterol and triglycerides were similar to that observed in the YC group (Fig. 3A). In both the AP and MP groups, the levels of high-density lipoprotein (HDL) were effectively restored to normal, similar to what was found in the YC group. Moreover, the low-density lipoprotein (LDL) levels were significantly decreased, reaching levels that were comparable to those in the YC group.

Fig. 3
figure 3

Effect of probiotics on the blood lipid profile of elderly mice receiving supplementation. AP: aged control group, YC: young control group, MP: middle-aged group and OC: old control group. (A) Measurement of serum cholesterol and triglyceride concentrations in elderly mice administered probiotics as opposed to their respective control cohorts. (n = 5–6 mice in each group) (B) Measurement of serum HDL and LDL concentrations in elderly mice administered probiotics as opposed to their respective control cohorts. (n = 5–6 mice in each group) The results shown in this figure are expressed as the mean ± SEM. An one-way ANOVA was used to analyze the comparisons of more than two groups followed by Tukey’s multiple comparison tests. P < 0.05 *, P < 0.01 **, and P < 0.001 ***

Full size image

The administration of probiotic supplements reduced liver inflammation in aged mice

Elderly experience changes in lipid metabolism that lead to increased levels of lipids, cholesterol, and triglycerides in their blood. These elevated lipid levels can accumulate in different organs, especially resulting in ectopic lipid deposition in the liver. In the liver tissue of OC mice, there was a significant increase in fibrosis regions and leukocyte infiltration compared to the YC group. (Upper panel, Fig. 4A). The liver tissue of YC mice demonstrated a typical distribution pattern of glycogen accumulation (Upper panel, Fig. 4A). The fibrotic regions showed significant improvement in mice from the AP and MP groups, where hepatocytes typically store glycogen (Upper panel, Fig. 4A). Additionally, we observed lipid accumulation in the livers of OC mice through oil red O staining, which was significantly mitigated by the administration of probiotics (Lower panel, Fig. 4A). In particular, polyunsaturated fatty acids are susceptible to oxidative destruction possibly caused by reactive oxygen species (ROS)-mediated lipid peroxidation. The OC group showed a significant increase in lipid peroxidation measured by malondialdehyde (MDA) levels and reduction of superoxide dismutase (SOD) activity (Fig. 4B). The probiotic combination resulted in a significant decrease in MDA, SOD (Fig. 4B), An excess of lipid storage in the liver is often associated with the beginning of liver inflammation. In this context, we measured the concentrations of the proinflammatory cytokines IL-1β and TNF-α in the liver, finding a significant reduction in both the MP and AP groups (Fig. 4C). Reduction in SOD level was inversely linked to increased levels of the liver enzymes AST and ALT, as illustrated in Fig. 4D in OC mice while liver enzymes were restored in AP and MP group. This suggests that the probiotic blend may help reduce liver inflammation by lowering oxidative stress and liver damage.

Fig. 4
figure 4

Potential probiotics restore normal lipid levels in the liver, reduce oxidative stress, and alleviate inflammation in elderly mice. (A) The upper panel depicts the presence of excessive lipid accumulation, while the lower panel shows the staining of lipid accumulation in liver tissue of elderly mice that were administered probiotics, as compared to their respective control groups. (n = 3–5 section from 5 mice in each group) (B) Quantitative of anti-oxidant SOD level and oxidative stress MDA level. (n = 5–6 mice in each group) (C) Quantitative of proinflammatory cytokine level (D) Quantitative of liver enzymes AST and ALT level in liver tissue of elderly mice received probiotics in comparison to control groups (n = 5 mice in each group). The results shown in this figure are expressed as the mean ± SEM. A one-way ANOVA was used to analyze the comparisons of more than two groups followed by Tukey’s multiple comparison tests. P < 0.05 *, P < 0.01 **, and P < 0.001 ***

Full size image

Supplementing of combination probiotics improve age-related cognitive decline in elderly mice

Aging is marked by high levels of oxidative stress and neuroinflammation, both of which are critical factors that lead to a decrease in cognitive functions like learning and memory. The administration of a combined probiotic to elderly mice in the MP and AP groups led to a significant elevation in the brain levels of the antioxidant enzyme superoxide dismutase (SOD). These levels were comparable to the concentrations observed in the YC group (Fig. 5A). Oxidative stress analysis in the brain revealed a significant decrease in brain MDA levels in older mice that received a combined probiotic intervention (Fig. 5A). Furthermore, the YC group showed increased levels of proinflammatory cytokines, particularly IL-1β and TNFα. The combined probiotic treatment significantly lowered these levels (Fig. 5B). As shown in Fig. 5C, the results indicate that the use of a combination of probiotics greatly enhanced neurogenesis in the hippocampus of aged mice in both the MP and AP groups, effectively matching their neurogenesis levels with those of the YC group. To further assess how these combined probiotics might enhance cognitive functions like learning and memory in older mice, a Morris Water Maze test was performed. The findings showed that older mice in the MP and AP groups maintained their cognitive abilities after receiving the combined probiotic. The mRNA expression of app and tau were determined, which showed the lower levels in elderly probiotics-treated mice (Fig. 5E). Taken together, these results suggest that probiotic could prevent age-associated memory impairment involving in reduction of oxidative stress and inflammation in elderly mice.

Fig. 5
figure 5

Potential probiotics restore normal levels of brain oxidative stress and reduce inflammation, thereby enhancing cognitive function in elderly mice. AP: aged control group, YC: young control group, MP: middle-aged group and OC: old control group. (A) Quantitative of anti-oxidant SOD level and oxidative stress MDA level. (n = 3 mice in each group) (B) Comparison of the quantitative levels of proinflammatory cytokines in the brain tissue of elderly mice that were given probiotics with those of the control groups. (n = 3 mice in each group). (C) Representative of histological hippocampus from elderly mice received probiotics in comparison to their control groups, 20x magnification. (D) The cognitive recognition assessment through the Morris water maze spatial test yielded a specific evaluation score (n = 5 mice in each group in duplicated). (E) Quantitative of neurogenesis-related genes expression level in elderly mice received probiotics in comparison to control groups (n = 3–4 mice in each group). The results shown in this figure are expressed as the mean ± SEM. An one-way ANOVA was used to analyze the comparisons of more than two groups followed by Tukey’s multiple comparison tests. P < 0.05 *, P < 0.01 **, and P < 0.001 ***

Full size image

Discussion

As individuals age, they often experience a decline in physical abilities and biological functions, usually associated with increased oxidative stress and inflammation. Therefore, anti- aging research investigations focus on alleviating such factors to reduce the risk of diseases such as Alzheimer. We show in our study, a combination of these two strains reduces oxidative stress and inflammation of the brain and liver while maintaining gut integrity in elderly mice.

The gut is crucial for probiotic development, and monitoring gut changes such as gut structure can assess probiotic effects. Research indicates that aged mice, whether treated with D-galactose or naturally aged, show gut structural abnormalities, such as fewer and shorter intestinal crypts, reduced goblet cells, and lower intestinal stem cell activity. These changes suggest a strong connection between gut structure and the aging process [6, 14]. In this study, our combination strains of probiotics, L. paracasei MSMC39-1 and B. animalis MSMC83. demonstrated an inhibitory effect on the characteristics of colonic aging in older mice by enhancing the quantity of crypts, despite a reduction in their relative size. This phenomenon is likely attributable to an improved turnover of epithelial cells or a decrease in apoptosis. Previous investigations have reported that the administration of probiotics to aged mice resulted in an increased number of crypts without a significant increase in their length, indicating a healthier intestinal condition in these aged subjects [15, 16] which are consistent with our results. Aged mice experience gut architecture degradation linked to inflammatory responses in the gastrointestinal tract. This study found that a specific probiotic formulation positively impacts intestinal health in aged mice by reducing expression of inflammation-related molecules in gut tissues, correlating with decreased gut inflammation. Previous research supports the role of probiotics in alleviating intestinal inflammation and normalizing crypt length and quantity [17, 18]. However, this study did not establish a clear relationship between inflammation and gastrointestinal injury, which may indicate gut structure is being altered while gut inflammation is increased.

Our results demonstrated that the probiotics administered did not have a significant impact on the expression of tight junction proteins. This may be due to probiotics protecting the gut barrier through other mechanisms. As an example, research carried out by Caballero-Franco et al. revealed that the administration of probiotics causes increased mucus production thus enhancing mucus protection of the body [19]. Other study, Forsyth et al., showed that probiotics effectively reduce gut inflammation and protect the gut barrier in a colitis animal model [20]. Additionally, Peng et al. found that butyrate, improves that function through directing the growth and shaping the gut epithelium [21]. Elevated lipid or cholesterol levels in elderly mice can lead to hepatic issues, but our combination of probiotics can reduce lipid accumulation possibly by boosting antioxidant levels, and modulate inflammation. Incorporating probiotics into the diets of aged mice improves gastrointestinal health and lipid metabolism, leading to lower cholesterol, triglycerides, HDL, and LDL levels. Other studies suggest these advantages were found to be linked to activity of bile salt hydrolase and regulatory mechanisms controlling metabolism pathways [22, 23]. In addition to their promising effects on metabolic profile probiotic supplementation has also been shown to improve the cognitive effect in aged mice. Similarly, research shows that probiotics lower neuroinflammatory markers IL-1β, IL-6, and TNF-α [24] reduce oxidative stress markers such as MDA, and increase SOD and catalase [25], in the brain, leading to improved cognitive health.

We found that probiotic supplementations could modulate the inflammatory and oxidative stress state in the brain of aged mice. In this case, beneficial effects of probiotics on neurogenesis that were seen in hippocampus and cognitive function are at least meditated by decreasing levels of tau and app mRNA expression. Together with our results, it may be speculated that probiotics could modulate cognitive function by reducing neuroinflammation and oxidative stress.

Conclusion

In conclusion, these beneficial effects are attributed to a decrease in intestinal inflammation through decreased expression of proinflammatory cytokines that accompanied by an increased antioxidative efficiency and an improvement of the metabolic profile as well as improved cognitive function in aged mice. It implies potential synergies of probiotics to counteract age-related diseases. In the future, microbiome and metabolome analysis will be necessary to gain a deeper understanding the interaction of our combination probiotics and gut environment and how their metabolites involve in reducing gut inflammation in aged mice.

Data availability

Data is provided within the manuscript or supplementary information files.

References

  1. Zhao H, Wu L, Yan G, Chen Y, Zhou M, Wu Y, Li Y. Inflammation and tumor progression: signaling pathways and targeted intervention. Signal Transduction and Targeted Therapy; 2021. p. 6.

  2. Hajam Y, Rani R, Ganie S, Sheikh T, Javaid D, Qadri S, Pramodh S, Alsulimani A, Alkhanani M, Harakeh S, Hussain A, Haque S, Reshi M. 2022. Oxidative Stress in Human Pathology and Aging: Molecular Mechanisms and Perspectives. Cells, 11.

  3. Sochocka M, Koutsouraki E, Gąsiorowski K, Leszek J. Vascular oxidative stress and mitochondrial failure in the pathobiology of Alzheimer’s disease: a new approach to therapy. CNS Neurol Disord Drug Target. 2013;12(6):870–81.

    Article  CAS  Google Scholar 

  4. Klenerman D, Meng J, Zhang Y, Saman D, De S, Sang J, Jiang K, Humphrey J, Julian L, Hidari E, Floto R, Lee S, Bryant C, Benesch J, Ye Y. Hyperphosphorylated Tau self-assembles into amorphous aggregates eliciting TLR4-dependent inflammatory responses. Nat Commun. 2022;13:2692.

  5. Bellary S, Kyrou I, Brown J, Bailey C. Type 2 diabetes mellitus in older adults: clinical considerations and management. Nat Reviews Endocrinol. 2021;17:534–48.

    Article  Google Scholar 

  6. Soty M, Gautier-Stein A, Rajas F, Mithieux G. Gut-Brain glucose signaling in energy homeostasis. Cell Metabol. 2017;25(6):1231–42.

    Article  CAS  Google Scholar 

  7. Huang S, Chen L, Wang M, Hsu C, Chan C, Li J, Huang H. 2018. Lactobacillus paracasei PS23 Delays Progression of Age-Related Cognitive Decline in Senescence Accelerated Mouse Prone 8 (SAMP8) Mice. Nutrients, 10.

  8. Cheng L, Chou P, Hou A, Huang C, Shiu W, Wang S. Lactobacillus paracasei PS23 improves cognitive deficits via modulating the hippocampal gene expression and the gut microbiota in D-galactose-induced aging mice. Food & function; 2022.

  9. Corpuz H, Ichikawa S, Arimura M, Mihara T, Kumagai T, Mitani T, Nakamura S, Katayama S. 2018. Long-Term Diet Supplementation with Lactobacillus paracasei K71 Prevents Age-Related Cognitive Decline in Senescence-Accelerated Mouse Prone 8. Nutrients,10.

  10. Wang S, Ahmadi S, Nagpal R, Jain S, Mishra S, Kavanagh K, Zhu X, Wang Z, McClain D, Kritchevsky S, Kitzman D, Yadav H. Lipoteichoic acid from the cell wall of a heat killed Lactobacillus paracasei D3-5 ameliorates aging-related leaky gut, inflammation and improves physical and cognitive functions: from C. elegans to mice. GeroScience. 2019;42:333–52.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Xia C, Cao X, Cui L, Liu H, Wang S, Chen T. Anti-aging effect of the combination of bifidobacterium longum and B. animalis in a d-galactose-treated mice. J Funct Foods. 2020;69:10398.

  12. Ladda B, Jantararussamee C, Pradidarcheep W, Kasorn A, Matsathit U, Taweechotipatr M. 2023. Anti-Inflammatory and Gut Microbiota Modulating Effects of Probiotic Lactobacillus paracasei MSMC39-1 on Dextran Sulfate Sodium-Induced Colitis in Rats. Nutrients, 15.

  13. Vitheejongjaroen P, Kasorn A, Puttarat N, Loison F, Taweechotipatr M. Bifidobacterium animalis MSMC83 improves oxidative stress and gut microbiota in D-Galactose-Induced rats. Antioxidants. 2022;11. https://doiorg.publicaciones.saludcastillayleon.es/10.3390/antiox11112146.

  14. Wang J, Hu J, Song Y, Yin J, Wang Y, Peng B, Zhang B, Liu J, Dong L, Wang S. 2022. 2′-Fucosyllactose Ameliorates Oxidative Stress Damage in d-Galactose-Induced Aging Mice by Regulating Gut Microbiota and AMPK/SIRT1/FOXO1 Pathway. Foods, 11.

  15. Khalesi S, Bellissimo N, Vandelanotte C, Williams S, Stanley D, Irwin C. A review of probiotic supplementation in healthy adults: helpful or hype. Eur J Clin Nutr. 2018;73:24–37.

    Article  PubMed  Google Scholar 

  16. Matsumoto M, Kurihara S, Kibe R, Ashida H, Benno Y. Longevity in mice is promoted by Probiotic-Induced suppression of colonic senescence dependent on upregulation of gut bacterial polyamine production. PLoS ONE 2011;6(8):e23652.

  17. Phillippi D, Daniel S, Nguyen K, Penaredondo B, Lund A. 2022. Probiotics Function as Immunomodulators in the Intestine in C57Bl/6 Male Mice Exposed to Inhaled Diesel Exhaust Particles on a High-Fat Diet. Cells, 11.

  18. Kato S, Hamouda N, Kano Y, Oikawa Y, Tanaka Y, Matsumoto K, Amagase K, Shimakawa M. Probiotic bifidobacterium bifidum G9-1 attenuates 5‐fluorouracil‐induced intestinal mucositis in mice via suppression of dysbiosis‐related secondary inflammatory responses. Clin Exp Pharmacol Physiol. 2017;44:1017–25.

    Article  CAS  PubMed  Google Scholar 

  19. Dai C, Zhao D, Jiang M. VSL#3 probiotics regulate the intestinal epithelial barrier in vivo and in vitro via the p38 and ERK signaling pathways. Int J Mol Med. 2011;29(2):202–8.

    PubMed  Google Scholar 

  20. Forsyth C, Farhadi A, Jakate S, Tang Y, Shaikh M, Keshavarzian A. Lactobacillus GG treatment ameliorates alcohol-induced intestinal oxidative stress, gut leakiness, and liver injury in a rat model of alcoholic steatohepatitis. Alcohol. 2009;43(2):163–72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Peng L, Li Z, Green R, Holzman I, Lin J. Butyrate enhances the intestinal barrier by facilitating tight junction assembly via activation of AMP-activated protein kinase in Caco-2 cell monolayers. J Nutr. 2009;139(9):1619–25.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Kumar R, Grover S, Batish V. Hypocholesterolaemic effect of dietary inclusion of two putative probiotic bile salt hydrolase-producing Lactobacillus plantarum strains in Sprague–Dawley rats. Br J Nutr. 2010;105:561–73.

    Article  PubMed  Google Scholar 

  23. Park J-E, Oh S-H, Cha Y-S. Lactobacillus plantarum LG42 isolated from Gajami sik-hae decreases body and fat pad weights in diet-induced obese mice. J Appl Microbiol [online]. 2014;116(1):145–56.

    Article  CAS  PubMed  Google Scholar 

  24. Azm S, Djazayeri A, Safa M, Azami K, Ahmadvand B, Sabbaghziarani F, Sharifzadeh M, Vafa M. Lactobacilli and bifidobacteria ameliorate memory and learning deficits and oxidative stress in β-amyloid (1–42) injected rats. Appl Physiol Nutr Metab. 2018;43(7):718–26.

    Article  Google Scholar 

  25. Davari S, Talaei S, Alaei H, Salami M. Probiotics treatment improves diabetes-induced impairment of synaptic activity and cognitive function: behavioral and electrophysiological proofs for microbiome–gut–brain axis. Neuroscience. 2013;240:287–96.

Download references

Acknowledgements

Department of Microbiology the Srinakharinwirot University, Department of Anatomy, The Center of Excellence in Probiotics and Animal House, Srinakharinwirot University.

Funding

Fundamental Fund, Thailand Science Research and Innovation (grant number 028/2567), Faculty of Medicine and HRH Princess Maha Chakri Sirindhorn Medical Center, Srinakharinwirot University (grant number149/2566, grant number 505/2566), and Excellence Center in Probiotics Srinakharinwirot University, grant number 055/2567).

Author information

Authors and Affiliations

  1. Innovative Anatomy Program, Department of Anatomy, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand

    Kaikwa Wuttisa

  2. Center of Excellence in Probiotics, Srinakharinwirot University, Bangkok, 10110, Thailand

    Pol Sookpotarom, Benjamaporn Poopan, Chantanapa Chantarangkul, Praewpannarai Jamjuree & Malai Taweechotipatr

  3. Doctor of Medicine Program, Faculty of Medicine, Srinakharinwirot University, Bangkok, 10110, Thailand

    Pol Sookpotarom & Malai Taweechotipatr

  4. Futuristic Science Research Center, School of Science, Walailak University, Thasala, Nakhon Si Thammarat, 80160, Thailand

    Jirapat Namkeaw & Thiranut Jaroonwitchawan

  5. Research Excellence Center for Innovation and Health Products (RECIHP), School of Allied Health Sciences, Walailak University, Nakhon Si Thammarat, 80160, Thailand

    Thiranut Jaroonwitchawan

  6. Clinical Research Center, Faculty of Medicine, Srinakharinwirot University, Ongkharak, Nakhon Nayok, 26120, Thailand

    Malai Taweechotipatr

Authors
  1. Kaikwa Wuttisa
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Pol Sookpotarom
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Benjamaporn Poopan
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Chantanapa Chantarangkul
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Praewpannarai Jamjuree
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. Jirapat Namkeaw
    View author publications

    You can also search for this author inPubMed Google Scholar

  7. Thiranut Jaroonwitchawan
    View author publications

    You can also search for this author inPubMed Google Scholar

  8. Malai Taweechotipatr
    View author publications

    You can also search for this author inPubMed Google Scholar

Contributions

Wuttisa K performed the writing of the original draft, designed the methodology, and contributed to the study investigation and initial prepared Figs. 1, 2, 3, 4 and 5, and curation, formal analysis, and visualization of the data; Sookpotarom Pol: contributed to the study investigation; Poopan B: contributed to the study investigation; Chantarangkul C: contributed to the study investigation; Jamjuree P: contributed to the study investigation; Namkeaw J: contributed to the study investigation, formal analysis and prepared figure 1; Jaroonwitchawan T: performed the writing, prepared Figs. 1, 2, 3, 4 and 5, revised the manuscript and formal analysis, contributed to the study conception and design; Taweechotipatr M; C secured funding for this study, supervised the experiments, contributed to the study conception and design. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Thiranut Jaroonwitchawan or Malai Taweechotipatr.

Ethics declarations

Ethics approval and consent to participate

Institutional review board statement: This study was approved by the Institutional Review Board of Srinakharinwirot University (No. SWU-A-015-2564). Institutional animal care and use committee statement: All protocols were carried out in accordance with relevant guidelines and regulations.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Material 1

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wuttisa, K., Sookpotarom, P., Poopan, B. et al. The potential of novel gut microbiota supplement in mitigating gut inflammation, alleviating oxidative stress linked to aging, and improving cognitive function in aged mice. BMC Complement Med Ther 25, 137 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12906-025-04881-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12906-025-04881-3

Keywords

BMC Complementary Medicine and Therapies

ISSN: 2662-7671

Contact us