TWI900986B - Faecalibacterium prausnitzii, postbiotics, and use thereof - Google Patents

Abstract

The present invention relates to a Faecalibacterium prausnitzii, postbiotics, and use thereof. The Faecalibacterium prausnitziiis selected from a group consisting of NCKUMT001 strain and NCKUMT002 strain. The postbiotics of the Faecalibacterium prausnitziiis also existed in a conditioned medium thereof and comprises extracellular vesicles and short chain fatty acids. The Faecalibacterium prausnitziiand the postbiotics of the Faecalibacterium prausnitziihave effects of protecting intestinal of a subject, ameliorating inflammation and regulation expression of lipid metabolism-related genes.

Description

Clostridium tenella, its postbiotics and uses thereof

The present invention relates to Clostridium tenella, its postbiotics and uses thereof.

Probiotics broadly refer to bacteria that are beneficial to the body. They are currently believed to contribute to intestinal health and even the immune system. Recent studies have shown that postbiotics, the metabolic products of probiotics, are crucial molecules for achieving their beneficial effects. Postbiotics include extracellular vesicles, organic acids, short-chain fatty acids, exopolysaccharides, lipopolysaccharides, probiotic lysates, vitamins, and amino acids. Postbiotics can enhance the growth of probiotics or inhibit the growth of harmful microorganisms, thereby improving intestinal flora. They also exhibit biological activities such as reducing inflammation, providing antioxidant properties, and alleviating hyperglycemia. Furthermore, as an increasing number of different bacteria are proven to contribute to health, the term "probiotics" encompasses an ever-expanding range of bacterial species.

However, probiotics of different strains, as well as probiotics of the same strain but different isolates, do not all provide the same beneficial effects on organisms. Therefore, isolating probiotics with distinct bioactivities has always been an important research and development direction for researchers in related fields.

The present invention relates to a Clostridium prausnitzii strain, a postbiotic thereof, and uses thereof. The Clostridium prausnitzii strain is selected from the group consisting of strains NCKUMT001 and NCKUMT002, wherein the accession number of strain NCKUMT001 is NITE BP 03861, and the accession number of strain NCKUMT002 is NITE BP 03862.

The present invention also relates to a postbiotic for Clostridium tenella, comprising a postbiotic of at least one of Clostridium tenella NCKUMT001 and Clostridium tenella NCKUMT002.

The present invention also relates to a composition containing postbiotics from Clostridium tenella NCKUMT001 or NCKUMT002.

Furthermore, the present invention also relates to the use of Clostridium tenella and Clostridium tenella postbiotics in preparing compositions for anti-inflammation, intestinal protection, improved lipid metabolism, protection against damage induced by intestinal infections, alleviation of intestinal infection symptoms, and reduction of intestinal infection mortality. The compositions comprise at least one of Clostridium tenella and Clostridium tenella postbiotics, and the compositions can be used to inhibit the expression of genes associated with inflammation, maintain intestinal cell integrity, and promote the expression of genes associated with lipid metabolism.

In one embodiment, the postbiotics of Clostridium tenella include the conditioned medium, extracellular vesicles, or a combination thereof of the NCKUMT001 strain or the NCKUMT002 strain.

In one embodiment, the postbiotics of Clostridium tenella comprise short-chain fatty acids.

In one embodiment, the composition containing postbiotics from Clostridium tenella NCKUMT001 or NCKUMT002 further comprises at least one of a food or pharmaceutically acceptable carrier, excipient, adjuvant, antioxidant, or food additive.

In one embodiment, the inflammation-related genes include serum amyloid A1, interleukin-1β, macrophage inflammatory protein-2 (MIP-2), and interleukin-8.

In one embodiment, the lipid metabolism-related gene comprises the peroxisome proliferator-activated receptor γ gene.

In one embodiment, the intestinal bacterial metabolic genes include: bile salt hydrolase (BSH) gene, hydroxysteroid dehydrogenase (HSDH) gene and butyrate transaminase (But) gene.

In one embodiment, Clostridium tenella postbiotics reduce intestinal inflammation and damage to intestinal cell close connections caused by pathogen infection, protect against damage induced by intestinal infection, alleviate symptoms of intestinal infection, and reduce mortality from intestinal infection.

In one embodiment, the Clostridium tenella postbiotic promotes cell tight junction protein 1 (CLDN1) gene expression.

Thus, the Clostridium tenella and its postbiotics, such as its secretosomes, of the present invention have multiple functions, including reducing inflammatory responses, protecting intestinal health, regulating lipid metabolism, protecting against damage induced by intestinal infections, alleviating symptoms of intestinal infections, and reducing mortality from intestinal infections.

In order to provide a more complete and clear disclosure of the effects that can be achieved by the technical means of the present invention, a detailed description is given below, and please refer to the disclosed drawings.

1. Isolation of Clostridium tenella strains

The NCKUMT001 and NCKUMT002 strains tested in this example were isolated from human fecal samples. They were cultured in BHIS (Brain-Heart Infusion Broth) at 37°C in an anaerobic environment.

Furthermore, 16S rRNA sequencing of strains NCKUMT001 and NCKUMT002 revealed 98-99% similarity between their 16S rRNA sequences and those of standard strains of Clostridium tenellae. Therefore, strains NCKUMT001 and NCKUMT002 were identified as Clostridium tenellae. The standard strains of Clostridium tenellae used in this experiment were Clostridium tenellae ATCC27768 (hereinafter referred to as ATCC27768) and Clostridium tenellae APC918/95b (hereinafter referred to as APC918/95b), respectively. The ATCC27768 strain is registered in the American Type Culture Collection as ATCC27768, and the APC918/95b strain is registered in the Japan Collection of Microorganisms as JPMC-918/95b. The number of the strain is JCM39207. See the first figure, which shows Gram-stained images of strains NCKUMT001, NCKUMT002, and ATCC 27768. All three strains are Gram-positive and exhibit similar growth patterns.

Next, genomic DNA was extracted from strains NCKUMT001 and NCKUMT002, and polymerase chain reaction (PCR) was performed using primers specific for Clostridium tenella type I to determine the genotypes (Phylogroups) of the two strains. See the agar electrophoresis image in Figure 2 (A), which shows that strains ATCC27768 and NCKUMT002 belong to type I, while strain NCKUMT001 belongs to type II. Figure 2 (B) shows the results of random amplified polymorphic DNA PCR analysis. This assay uses primers targeting different bacterial genes, conducts PCR, and then subjects the products to agar gel electrophoresis to observe the DNA fragments amplified by each strain. Figure 2 (B) shows that the DNA fragments amplified by strains NCKUMT002 and ATCC 27768 are similar, while the DNA fragments amplified by strain NCKUMT001 differ from those of the aforementioned two strains, confirming that strain NCKUMT001 and the other two strains are distinct genotypes.

Both strains NCKUMT001 and NCKUMT002 have been deposited with the NITE Patent Microorganisms Depositary National Institute of Technology and Evaluation in Japan. The deposit number for strain NCKUMT001 is NITE BP 03861, and the deposit number for strain NCKUMT002 is NITE BP 03862.

2. Isolation and Analysis of Nutritional Secretions of Clostridium tenella

Seed cultures of strains NCKUMT001 and NCKUMT002 were cultured in mBHIS (modified Brain-Heart Infusion Broth) medium under anaerobism for 48 hours. The culture medium was centrifuged at 4000 rpm for 10 minutes, and the supernatant was collected. This supernatant is the conditioned medium of Clostridium tenella and is subsequently referred to as "supplemental nutrient solution (SUP)." Figure 1 (B) shows an electron microscopic photograph of extracellular vesicles (EVs) observed in the SUP of ATCC 27768 strain. Also, see Table 1. ATCC 27768 can be measured using the particle concentration method and the qNano instrument. The concentrations of exosomes and proteins in the nutrient secretions (SUP) of strains 27768, NCKUMT001, and NCKUMT002 are shown in Table 1. The results show that all three strains can produce exosomes and that the exosomes contain proteins.

Table 1 strain ATCC 81047 NCKUMT001 NCKUMT002 Bacterial concentration (CFU/ml) 2.85X10 ⁷ 2.9X10 ⁵ 3.5X10 ⁶ Exosome particle number (/ml) 2.03X1012 5.92X1011 8.27X1011 The number of cytosomes per bacterium (/CFU) 1.4X10 ⁵ 4.9X10 ⁵ 4.2X10 ⁴ Exosome protein concentration (mg/ml) 0.19 0.21 0.22

Please see the third figure, which shows the results of quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) analysis of the relative expression levels of short-chain fatty acids (SCFAs) in the three strains. The results show that the SCFAs in strain NCKUMT001 are higher than those in strains ATCC27768 and NCKUMT002.

Please see Table 2 for the analysis of short-chain fatty acid concentrations in the nutritional secretions (SUP) of strains NCKUMT001 and NCKUMT002. The butyrate content in the SUP of both strains was significantly higher than that in the blank culture medium. Furthermore, the formic acid, ethyl acetate, and acetic acid content in the SUP of strain NCKUMT001 were also significantly higher than those in the blank culture medium.

Table 2 Formic acid (μM) Acetic acid (μM) Propionic acid (μM) Butyrate (μM) Valeric acid (μM) Blank culture medium 564 18472 40 68 0.6 NCKUMT001 1141 27216 61 1870 0.9 NCKUMT002 661 19680 50 1232 0.5

Table 3 shows the minimum inhibitory concentration (MIC) test results for different antibiotics against NCKUMT001 and NCKUMT002 strains. The antibiotics tested included Ertapenem, Imipenem, Clindamycin, Metronidazole, and Ampicillin-Sulbactum. The results showed that both strains were not resistant.

Table 3 Ertapenem Imipenem Clindamycin Metronidazole Ampicillin-Sulbactum NCKUMT001 0.125 0.25 0.25 0.125 < 0.0625 NCKUMT002 0.125 2 0.125 0.125 < 0.0625

3. Bioactivity Test of NCKUMT001 Strain and NCKUMT001 Strain

(1) Inhibit inflammatory response

In this study, human colorectal cancer cell line (HT29 cell line) was used for testing. First, 100 mg/mL of lipopolysaccharide (LPS) and 10 Cells were stimulated with 100 ng/mL of interferon-γ (IFN-γ). One hour after stimulation, a nutrient secretion (SUP) containing 10% (v/v) NCKUMT001 was added to the cell culture medium. The cells were cultured for an additional hour, harvested, and RNA was extracted. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was then used to analyze the expression of interleukin-8 (IL-8) mRNA in the cells. As shown in Figure 4 (A), the IL-8 expression levels in the SUP-supplemented groups were significantly lower than those in the SUP-unsupplemented groups. These results indicate that the nutrient secretion (SUP) of the NCKUMT001 strain has the effect of reducing the expression of inflammatory substances.

Please also refer to Figure 4 (B), which shows differences in IL-8 mRNA expression in HT29 cells after co-stimulation with LPS and IFN-γ. One hour after stimulation, 5% to 40% (v/v) of the NCKUMT001 strain's nutrient secretion (SUP) was added to the cells. Figure 4 (B) shows that IL-8 expression in the group supplemented with 5% (v/v) SUP was significantly lower than in the group without SUP. Furthermore, as the proportion of SUP added increased, IL-8 expression decreased, demonstrating a dose-dependent effect.

Furthermore, in Figure 4 (C), HT29 cells were co-stimulated with LPS and IFN-γ and then co-cultured with a culture medium containing 10% (v/v) NCKUMT001 bacteria. The expression of IL-8 mRNA in each cell group was observed. The low-concentration culture medium group had an absorbance of 1 at 600 nm (O.D. 600), while the high-concentration culture medium group had an O.D. 600 of 2. As shown in Figure 4 (C), IL-8 expression in both culture medium-supplemented groups was lower than that in the unsupplemented group, indicating that the culture medium containing NCKUMT001 bacteria did not promote inflammation and even had a slightly inhibitory effect.

(2) Protection against Clostridium difficile-induced damage

Next, the protective effects of NCKUMT001's nutrient secretions (SUP) or endosomes (EV) against the ATCC43255 strain of Clostridioides difficile (CDI) were tested. In this assay, HT29 cells were cultured for 6 hours in a culture supernatant containing 10% (v/v) C. difficile (CDI(SUP)). Then, 20% (v/v) of NCKUMT001's nutrient secretions (SUP) or endosomes (EV) were added, and the relative expression of cellular IL-8 mRNA was measured. As shown in Figure 5 (A), cell lines supplemented with either nutrient secretions (SUP) or endosomes (EVs) of the NCKUMT001 strain expressed significantly lower levels of IL-8 than those without such supplementation. This suggests that nutrient secretions (SUP) or endosomes (EVs) of the NCKUMT001 strain do have the effect of reducing the expression of inflammatory substances.

Figure 5 (B) shows the efficacy of Clostridium tenella in protecting cell membrane integrity. In the experiment, HT-29 cells were first cultured in the culture supernatant of Clostridium difficile (CDI(SUP)), and 20% (v/v) of nutrient secretions (SUP) or endosomes (EV) of the NCKUMT001 strain were added at the same time. The cells were then cultured for 2 hours. Then, the cells were stained with FITC (Fluorescein isothiocyanate). Fluorescent dye staining. If the cell membrane is damaged, the fluorescent dye will enter the cell and stain the cell. Collect the stained cells and measure the fluorescence intensity of each group of cells. The stronger the fluorescence intensity detected by the cells, the more serious the cell membrane damage. According to Figure 5 (B), the nutritional status of the NCKUMT001 strain is treated. The fluorescence intensity of cells in the groups treated with either nutrient secretions (SUP) or endosomes (EV) was significantly lower than that of the untreated group. Furthermore, the fluorescence intensity of the EV-treated group was the lowest among the three groups, indicating that both nutrient secretions (SUP) and endosomes (EV) of the NCKUMT001 strain can protect the cell membrane from cell membrane damage caused by Clostridium difficile.

Furthermore, Figure 5 (C) shows the expression of claudin (CLDN1) mRNA, a protein involved in tight cell junctions, in cells from each group under the same treatment. As shown in Figure 5 (C), cells treated with NCKUMT001 strain nutrient secretions (SUP) or endosomes (EV) showed significantly higher CLDN1 expression than those in the untreated group, indicating that C. tenella nutrient secretions (SUP) or endosomes (EV) increase CLDN1 expression and, in turn, promote tight cell junctions.

(3) Animal experimental models

In this study, C57BL/6J mice were used as an animal model to observe the effects of Clostridium tenella on their intestinal health. Please refer to Figure 6 (A) for details. The start date of this experiment is defined as "Day 0," the day before the experiment is defined as "Day -1," the day after the experiment is defined as "Day 1," and so on. The mice were divided into the following groups:

(1) Blank control group: The experimental mice did not receive any treatment;

(2) CDI group: Five days before the start of the experiment, the experimental mice were given antibiotics orally (please provide the type and dosage of antibiotics used: Kanamycin 0.4 mg/mL; Gentamycin 0.0035 mg/mL; Colistin 0.057 mg/mL; Metronidazole 0.215 mg/mL; Vancomycin 0.045 mg/mL) to destroy the normal intestinal flora of the experimental mice. After the start of the experiment, the mice were infected with ¹⁰⁴ CFU/ml of Clostridium difficile spores.

(3) CDI+EV group: 5 days before the start of the experiment, the experimental mice were given 200uL of NCKUMT001 strain EVs daily at a concentration of 10 ¹¹ /ml. After infection with Clostridium difficile, 200uL of NCKUMT001 strain EVs were continued daily until the end of the experiment.

On day 3 after infection, surviving mice were sacrificed and their intestinal tissues were collected for subsequent analysis.

Please see Figure 6 (B) for the analysis of the survival rates of the experimental mice in each group. None of the experimental mice in the blank control group died at the end of the experiment, so their survival rate was 100%. On day 3 after infection, the survival rate of the experimental mice in the CDI group was 60%, while the survival rate of the experimental mice in the CDI+EV group was the same as the blank control group, at 100%. This result shows that the cytosomes of the NCKUMT001 strain can indeed prevent the death of experimental mice caused by infection with Clostridium difficile.

Figure 6 (C) shows the analysis of intestinal length in each group of experimental mice. In this experiment, the small intestine length of the experimental mice was measured. Compared with the blank control group, the large intestine length of the experimental mice in the CDI group was significantly decreased, while the intestinal length of the experimental mice in the CDI+EV group was significantly increased compared with the CDI group. This result shows that the exosomes of the NCKUMT001 strain can effectively alleviate intestinal damage caused by bacterial infection in mice.

Please refer to Figures 7 (A) and (B), which show the expression levels of serum amyloid A1 (SAA1) and interleukin-1β (IL-1β) in the intestinal tissues of experimental mice, respectively. According to the test results, the expression levels of SAA1 and IL-1β in the intestinal tissues of experimental mice in the CDI group were significantly higher than those in the blank control group. However, the expression levels of these two cytokines in the intestinal tissues of experimental mice in the CDI+EV group were significantly lower than those in the CDI group. This result shows that the cytosomes of Clostridium tenella effectively reduce the expression of inflammatory proteins in intestinal cells.

(IV) Clostridium tenella affects intestinal bacterial metabolism

The purpose of the following experiment was to test the biological activity of Clostridium tenella related to intestinal bacterial metabolism. This experiment utilized an in vitro fermentation system developed by the inventors of this case. Human intestinal bacteria were cultured within this system to form a bacterial community composition that simulated the human intestinal flora (hereinafter referred to as the intestinal flora composition). The expression of metabolic genes related to intestinal flora was observed under different conditions.

In this experiment, there were three groups: (1) blank control group, in which the intestinal flora composition in the in vitro fermentation system did not receive any treatment; (2) antibiotic group, 150 mg/mL clindamycin was added to the culture medium of the intestinal flora composition in the in vitro fermentation system and cultured together for 1 day; and (3) SUP group, in which 150 mg/mL clindamycin antibiotic and 10% (v/v) nutrient secretion (SUP) of the NCKUMT001 strain were added to the culture medium of the in vitro fermentation system for 1 day. After the experiment, DNA from the intestinal flora of each group was collected and analyzed using real-time quantitative polymerase chain reaction to detect and analyze the levels of genes related to bile acid and butyrate metabolism in the samples.

Please see Figure 8 (A), which shows the gene count analysis of intestinal bacteria in each group. Compared to the blank control group, the gene count of intestinal bacteria in the antibiotic-treated group decreased significantly. However, the gene count of intestinal bacteria in the SUP group was significantly higher than that in the antibiotic group. This result shows that the nutrient secretion (SUP) of the NCKUMT001 strain can mitigate the reduction of intestinal bacteria caused by antibiotics.

Please see Figure 8 (B), which shows the gene abundance analysis of secondary cholate dehydrogenase (hydroxysteroid dehydrogenase) in the intestinal flora of each group. Compared to the blank control group, the gene abundance of secondary cholate dehydrogenase in the intestinal flora of the antibiotic-treated group decreased significantly, but the gene abundance of secondary cholate dehydrogenase in the intestinal flora of the SUP group was higher than that of the antibiotic group.

Please refer to Figure 8 (C), which shows the analysis of butyryl-CoA:acetate CoA-transferase gene abundance in the intestinal flora of each group. Compared to the blank control group, the number of butyryl-CoA:acetate CoA-transferase genes in the intestinal flora of the antibiotic-treated group decreased significantly, but the intestinal flora of the SUP group was significantly higher than that of the antibiotic group.

The results in Figure 8 show that Clostridium tenella in this case can reduce the death of intestinal flora caused by antibiotic use, restore the metabolic function of intestinal bacteria damaged by antibiotic use, and promote the ability of intestinal flora to metabolize bile acid and short-chain fatty acids, thereby maintaining intestinal health and regulating intestinal inflammatory responses.

Please refer to Figure 9, which uses HT-29 cells to test the effect of Clostridium tenella nutrient secretion (SUP) on the protection of cells from C. difficile infection. The experiment includes four groups: (1) blank control group, in which cells did not receive any treatment; (2) CDI group, in which cells were infected with C. difficile at a multiplicity of infection (MOI) of 5; (3) SUP group, in which cells were infected with C. difficile and cultured in a medium containing 10% (v/v) (4) EV group, cells were infected with Clostridium difficile and cultured in culture medium containing 10% (v/v) EV of NCKUMT001 strain for 4 h.

The results in Figure 9 show that in cells infected with B. difficile, the expression of peroxisome proliferator-activated receptor-γ (PPAR-γ) decreased significantly, falling significantly below that of the blank control group. However, PPAR-γ levels in the SUP and EV groups increased significantly, even exceeding those of the blank control group. PPAR-γ is a key factor in regulating the human immune response. In addition to its ability to reduce inflammation, it also promotes fat metabolism and reduces body weight.

(5) Clostridium tenella regulates inflammatory response

This study was conducted using GES-1 gastric epithelial cells. The GES-1 gastric epithelial cells were first stimulated with lipopolysaccharide and then co-cultured with NCKUMT001 bacterial solution (hereinafter referred to as the "bacterial solution group") or NCKUMT001 exosomes (hereinafter referred to as the "EV group"). The IL-8 expression of cells in each group was observed one day after culture. The amount of NCKUMT001 added to the bacterial solution group was an MOI (multiplicity of infection) of 100, while the exosomes added to the EV group was an MOI of 10. ⁴ cell secretomes/cell; please see Figure 10. Figure 10 (A) shows the IL-8 concentration analysis results of the cell culture supernatant, and Figure 10 (B) shows the relative expression analysis results of IL-8 mRNA in cells of each group. According to Figures 10 (A) and (B), LPS stimulates cells to express IL-8, but the IL-8 mRNA expression and secretion levels of cells in the bacterial solution group and EV group are significantly reduced. That is, when cells are co-cultured with NCKUMT001 bacterial solution or co-cultured with NCKUMT001 cell secretomes, LPS-induced IL-8 is inhibited. The mRNA expression and IL-8 protein expression levels also indicate that the NCKUMT001 bacterial solution and its endosomes not only do not promote inflammatory responses, but may even significantly inhibit inflammatory responses.

In summary, the present invention discloses Clostridium tenella, its postbiotics, and uses thereof, comprising two novel isolates of Clostridium tenella. The nutritional cultures and secretions of these two strains have the ability to reduce gastrointestinal inflammation, maintain gastrointestinal health, and regulate lipid metabolism. In this embodiment, the nutritional secretions of the Clostridium tenella strains contain a variety of short-chain fatty acids that protect the integrity of intestinal cell membranes. Furthermore, the postbiotics of Clostridium tenella can promote the expression of genes related to lipid metabolism, thereby regulating lipid metabolism in an organism.

In summary, the Clostridium tenella, its postbiotics, and their uses of the present invention can achieve the intended efficacy through the disclosed embodiments. Furthermore, this invention, which had not been disclosed prior to filing this application, fully complies with the provisions and requirements of the Patent Law. Therefore, we hereby file this invention patent application in accordance with the law and earnestly request your review and grant of the patent. We are truly grateful for this.

However, the above description is only a preferred embodiment of the present invention and is not intended to limit the scope of protection of the present invention. Other equivalent changes or modifications made by persons skilled in the art based on the characteristic scope of the present invention should be considered as not departing from the design scope of the present invention.

without

Figure 1: Gram stain analysis, gene sequence analysis, and electron microscopy images of Clostridium tenellae secretomes. Figure 2: Phylogroup analysis of Clostridium tenellae. Figure 3: Analysis of short-chain fatty acid translocase expression in Clostridium tenellae. Figure 4: Analysis of inflammatory response substances regulated by nutritional secretions of Clostridium tenellae and Clostridium tenellae. Figure 5: Analysis of how nutritional secretions and secretions of Clostridium tenellae affect cellular inflammatory substances, membrane potential, and expression of the tight junction protein (CLDN1) gene. Figure 6: Analysis of the effects of postbiotics from Clostridium tenellae on survival and cecal length in animals subjected to intestinal infection. Figure 7: Analysis of the effects of postbiotics from Clostridium tenella on inflammatory response substances in experimental animals with intestinal infection. Figure 8: Analysis of the effects of postbiotics from Clostridium tenella on the abundance of intestinal bacteria, and the expression of genes involved in bile acid metabolism and short-chain fatty acid metabolism. Figure 9: Analysis of the effects of postbiotics from Clostridium tenella on the expression of genes related to lipid metabolism in experimental animals. Figure 10: Analysis of the effects of postbiotics from Clostridium tenella on gastric wall cell inflammation.

Depository: NITE Patent Microorganisms Depositary, National Institute of Technology and Evaluation, Japan Deposit Date: March 20, 2023 Deposit Number: The deposit number for strain NCKUMT001 is NITE BP 03861, and the deposit number for strain NCKUMT002 is NITE BP 03862.

Claims (10)

A Faecalibacterium prausnitzii strain that maintains intestinal cell integrity is selected from the group consisting of Faecalibacterium prausnitzii strains NCKUMT001 and NCKUMT002, wherein the NCKUMT001 strain has a deposit number of NITE BP 03861 and the NCKUMT002 strain has a deposit number of NITE BP 03862. A postbiotic for Faecalibacterium prausnitzii that maintains intestinal cell integrity is present in a conditioned medium of C. prausnitzii strain NCKUMT001 or C. prausnitzii strain NCKUMT002, wherein the NCKUMT001 strain has a deposit number of NITE BP 03861 and the NCKUMT002 strain has a deposit number of NITE BP 03862. The postbiotic according to claim 2, comprising extracellular vesicles of Clostridium tenella. The postbiotic as described in claim 2 comprises short-chain fatty acids. A composition for maintaining intestinal cell integrity comprises Clostridium tenella or at least one of its postbiotics, wherein the Clostridium tenella is selected from the group consisting of Clostridium tenella NCKUMT001 strain or Clostridium tenella NCKUMT002 strain, wherein the NCKUMT001 strain has the accession number NITE BP 03861, and the NCKUMT002 strain has the accession number NITE BP 03862; and wherein the postbiotic is present in a conditioned culture medium of the Clostridium tenella. The composition of claim 5, wherein the postbiotic comprises secretosomes of Clostridium tenella. The composition of claim 5, wherein the postbiotic comprises short-chain fatty acids. The composition of claim 5 comprises at least one of a food or a pharmaceutically acceptable carrier, excipient, adjuvant, antioxidant, or food additive. A use of Clostridium tenella or at least one of its postbiotics for preparing a composition for maintaining intestinal cell integrity, wherein the composition increases claudin expression, inhibits the expression of genes associated with gastrointestinal inflammation, maintains intestinal cell integrity, promotes the expression of genes associated with fatty acid metabolism, promotes the expression of genes associated with bile acid metabolism, and reduces antibiotic-induced intestinal bacterial death, wherein the Clostridium tenella is selected from the group consisting of Clostridium tenella NCKUMT001 strain and Clostridium tenella NCKUMT002 strain, wherein the NCKUMT001 strain has a registration number of NITE BP 03861 and the NCKUMT002 strain has a registration number of NITE BP 03861. The use as described in claim 9, wherein the inflammation-related gene comprises serum amyloid A1, interleukin-1β, and interleukin-8, and wherein the lipid metabolism-related gene comprises the peroxisome proliferator-activated receptor γ gene, the fatty acid metabolism-related gene comprises the short-chain fatty acid transferase gene, and the bile acid metabolism-related gene comprises the secondary bile acid transferase gene.