Faecalibacterium longum and its uses

Abstract

The present invention proposes the use of Faecalibacterium longum in ulcerative enteritis, and also provides a composition for treating and preventing ulcerative enteritis and related diseases thereof, including a medicine or an animal feed composition, etc. The Faecalibacterium longum was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on June 13, 2016, with a deposit number of CGMCC 1.5208, and a classification name of Faecalibacterium longum. Faecalibacterium longum has a significant therapeutic and preventive effect on ulcerative enteritis, which is specifically manifested in improving the apparent state of mice with ulcerative enteritis, reducing the disease activity index of mice, and reducing the inflammatory response of mice, which is significantly better than existing commercial ulcerative enteritis drugs.

Description

Technical Field

[0001] The present invention relates to the field of microorganisms, specifically, the present invention relates to long-dwelling Faecalibacterium and uses thereof, more specifically, the present invention relates to long-dwelling Faecalibacterium, uses of long-dwelling Faecalibacterium in preventing and treating ulcerative enteritis, uses of long-dwelling Faecalibacterium in preparing medicines or compositions, compositions, medicines, long-dwelling Faecalibacterium capsules and preparations. Background Art

[0002] Ulcerative colitis (UC) is a type of inflammatory bowel disease (IBD), a chronic intestinal inflammatory disease with an unknown pathogenesis. Clinical pathological studies have shown that the onset of UC is mainly related to individual susceptibility genes and the intestinal mucosal immune response triggered by intestinal microorganisms. Its symptoms are mainly persistent abdominal pain, diarrhea, and bloody stools, and the disease recurs, and can occur at any age.

[0003] At present, the main drugs used clinically for UC are salicylates, glucocorticoids, and immune preparations. Salicylic acid drugs can effectively inhibit the synthesis of prostaglandins and scavenge oxygen free radicals to relieve inflammatory responses. The most common salicylic acid drugs used clinically to treat UC are sulfasalazine (SASP), which is mainly used for patients with mild, moderate, and chronic UC. Glucocorticoids are the first choice for patients with severe or fulminant UC, such as betamethasone. Immunosuppressants such as cyclosporine can inhibit the production of T cell IL-2 and affect the progression of immune responses, thereby inhibiting UC.

[0004] The three existing types of drugs for UC can relieve UC to a certain extent, but they all have certain side effects. The side effects of salicylates are gastrointestinal reactions, headaches, reticulocytosis, sperm reduction, and rashes caused by allergic reactions, hepatotoxicity, leukopenia, anemia, etc. Glucocorticoids can cause metabolic disorders and water retention and other side effects. They can only be used as emergency medications and cannot be taken for a long time. Immunosuppressant treatment is highly dependent on drugs, has a long treatment cycle, and is prone to cause nephrotoxicity and secondary infections. It can only be used as an auxiliary treatment.

[0005] Currently, there is an urgent need in the art for an effective method and drug for treating and preventing UC with few side effects.

[0006] It is known that microorganisms have a certain connection with enteritis, and microorganisms will continue to be the focus of attention and research in enteritis to better understand their mechanism of action and their potential role in the treatment and prevention of enteritis. Summary of the invention

[0007] This application is based on the inventor's discovery and understanding of the following facts and problems:

[0008] The inventors of the present application screened a strain of Faecalibacterium longum (deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration on June 13, 2016, with a deposit number of CGMCC1.5208) (in the present application, the strain of Faecalibacterium longum is referred to as Faecalibacterium longum CM04-06). After conducting functional verification experiments, they were pleasantly surprised to find that Faecalibacterium longum CM04-06 was significantly better than existing commercial ulcerative enteritis drugs in inhibiting weight loss, increased disease activity index (DAI) and colon shortening in UC mice induced by dextran sulfate sodium (DSS).

[0009] To this end, in the first aspect of the present invention, the present invention proposes the use of Faecalibacterium longum in the treatment and prevention of enteritis. The long-living Faecalibacterium was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on June 13, 2016, with a deposit number of CGMCC 1.5208, and a classification name of: Faecalibacterium longum. The long-living Faecalibacterium CM04-06 according to the embodiment of the present invention has a significantly better therapeutic and preventive effect on DSS-induced UC mice in inhibiting weight loss, increased DAI and shortened colon symptoms in UC mice than existing commercial ulcerative enteritis drugs.

[0010] In the second aspect of the present invention, the present invention proposes the use of Faecalibacterium longum in the preparation of a medicine or a composition. The Faecalibacterium longum was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on June 13, 2016, with a deposit number of CGMCC 1.5208 and a classification name of: Faecalibacterium longum. The medicine or composition is used for at least one of the following: treating and preventing enteritis, inhibiting enteritis-induced weight loss, increased DAI, and improving intestinal changes.

[0011] According to an embodiment of the present invention, the above-mentioned use may further include at least one of the following additional technical features:

[0012] According to an embodiment of the present invention, the enteritis is ulcerative enteritis.

[0013] According to an embodiment of the present invention, the ulcerative enteritis is ulcerative colitis.

[0014] According to an embodiment of the present invention, the change in the intestine is shortening of colon tissue.

[0015] According to the embodiment of the present invention, Faecalibacterium longum CM04-06 has a good effect of inhibiting enteritis-induced weight loss, increased disease activity index (DAI) and improving enteritis-induced intestinal changes. The medicine and composition prepared using Faecalibacterium longum CM04-06 according to the embodiment of the present invention are effective in treating and preventing ulcerative colitis.

[0016] According to an embodiment of the present invention, the concentration of Faecalibacterium longum is 1.0×10 6 CFU / mL~1.0×10 9 CFU / mL. The drug prepared by using Faecalibacterium longifolium CM04-06 is more effective in treating, preventing or alleviating ulcerative enteritis, inhibiting enteritis-induced weight loss, increasing DAI and improving intestinal changes.

[0017] In the third aspect of the present invention, the present invention proposes a composition. According to an embodiment of the present invention, the composition comprises: the aforementioned Faecalibacterium longum, which was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on June 13, 2016, with a deposit number of CGMCC1.5208, and a classification name of: Faecalibacterium longum. The composition comprising Faecalibacterium longum CM04-06 according to an embodiment of the present invention is superior to existing ulcerative enteritis drugs in inhibiting enteritis-induced weight loss, increased DAI, and improving intestinal changes.

[0018] In the fourth aspect of the present invention, the present invention proposes a drug. According to an embodiment of the present invention, the drug comprises: Faecalibacterium longum, which was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration Committee on June 13, 2016, with a deposit number of CGMCC 1.5208, and a classification name of: Faecalibacterium longum. The drug containing Faecalibacterium longum CM04-06 according to an embodiment of the present invention is superior to existing ulcerative enteritis drugs in inhibiting enteritis-induced weight loss, DAI increase, and improving intestinal changes.

[0019] According to an embodiment of the present invention, the drug may further include at least one of the following additional technical features:

[0020] According to an embodiment of the present invention, the pharmaceutical composition further comprises: a pharmaceutically acceptable adjuvant.

[0021] According to an embodiment of the present invention, the pharmaceutically acceptable adjuvant includes at least one of a stabilizer, a wetting agent, an emulsifier, a binder, and an isotonic agent.

[0022] According to an embodiment of the present invention, the drug is in the form of at least one of tablets, granules, powders, capsules, solutions, suspensions, and freeze-dried preparations. Furthermore, the drug according to an embodiment of the present invention is convenient for administration and is suitable for maintaining the optimal activity of Faecalibacterium longiflorum.

[0023] According to the embodiments of the present invention, the dosage of the drug is not particularly limited, and in practical applications, it can be flexibly selected according to the health status of the subject.

[0024] According to some embodiments of the present invention, the medicine described in this embodiment is a compound probiotic.

[0025] In the fifth aspect of the present invention, the present invention proposes a long-lived Faecalibacterium capsule. According to an embodiment of the present invention, the long-lived Faecalibacterium capsule comprises: long-lived Faecalibacterium freeze-dried powder and maltodextrin, and the mass ratio of the long-lived Faecalibacterium freeze-dried powder to the maltodextrin is 1:1, wherein the long-lived Faecalibacterium was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on June 13, 2016, with a deposit number of CGMCC 1.5208, and a classification name of: Faecalibacterium longum. Capsules containing long-lived Faecalibacterium CM04-06 according to an embodiment of the present invention are superior to existing ulcerative enteritis drugs in inhibiting enteritis-induced weight loss, increased DAI, and improving intestinal changes.

[0026] According to an embodiment of the present invention, the above-mentioned Faecalibacterium longum capsule may further include at least one of the following additional technical features:

[0027] According to an embodiment of the present invention, the Faecalibacterium longum capsule further comprises: 0.5 g of Faecalibacterium longum freeze-dried powder and 0.5 g of maltodextrin.

[0028] According to an embodiment of the present invention, the lyophilized powder of Faecalibacterium longum is prepared by mixing the live bacteria content of 1 to 500×10 6 The CFU / mL of live Faecalibacterium longifolia was obtained by freeze-drying.

[0029] According to an embodiment of the present invention, the long-lived Faecalibacterium lyophilized powder includes: 0.8 to 1.2 parts by mass of the long-lived Faecalibacterium live bacteria, 1.8 to 2.2 parts by mass of lactose, 1.8 to 2.2 parts by mass of yeast powder, 0.8 to 1.2 parts by mass of peptone, 90 to 95 parts by mass of pure water and 0.3 to 1 parts by mass of vitamin C to maintain high activity.

[0030] In the sixth aspect of the present invention, the present invention proposes a long-living Faecalibacterium preparation. According to an embodiment of the present invention, the preparation comprises: 4 to 6 volumes of growth factor, 0.5 to 1.5 volumes of vitamin C, and 95 to 105 volumes of long-living Faecalibacterium fermentation liquid, wherein the long-living Faecalibacterium was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on June 13, 2016, with a deposit number of CGMCC 1.5208, and a classification name of: Faecalibacterium longum. The preparation containing the long-living Faecalibacterium CM04-06 according to the embodiment of the present invention is superior to existing ulcerative enteritis drugs in inhibiting enteritis-induced weight loss, increasing DAI, and improving intestinal changes.

[0031] According to an embodiment of the present invention, the above-mentioned Faecalibacterium longum preparation may further include at least one of the following additional technical features:

[0032] According to an embodiment of the present invention, the growth factor is a crude extract of milk growth factor.

[0033] According to an embodiment of the present invention, the growth factor is further at least one of a vitamin substance, a purine substance, and a pyrimidine substance.

[0034] According to an embodiment of the present invention, the growth factor is a crude milk growth factor extract obtained by sequentially mixing, centrifuging and ultrafiltering skim milk and casein.

[0035] According to an embodiment of the present invention, the fermented bacterial liquid of Faecalibacterium longum is obtained by subjecting the Faecalibacterium longum to anaerobic fermentation at 37° C. for 2 to 3 days.

[0036] According to an embodiment of the present invention, the Faecalibacterium longum preparation further comprises: maltodextrin.

[0037] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0039] Figure 1 is a neighbor-joining phylogenetic tree drawn using MegaX software for Faecalibacterium longum CM04-06 according to an embodiment of the present invention;

[0040] Figure 2 is a result diagram of the weight change of mice after Faecalibacterium longum CM04-06 intervened in mice according to an embodiment of the present invention; and

[0041] Figure 3 This is a result diagram of the change in DAI score of mice after Faecalibacterium CM04-06 intervened in mice according to an embodiment of the present invention. DETAILED DESCRIPTION

[0042] Embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and intended to be used to explain the present invention, but should not be understood as limiting the present invention.

[0043] The following examples screened out a strain of Faecalibacterium CM04-06 that can prevent and treat ulcerative enteritis through in vitro probiotic property evaluation. In order to verify the efficacy of the strain in treating and alleviating ulcerative enteritis in vivo, the inventors used an enteritis mouse model to carry out functional verification experiments. Dextran Sulfate Sodium (DSS) was used as an inducer to cause diarrhea in mice and induce intestinal inflammation to establish an enteritis mouse model. The DSS-induced mouse model is the most widely used animal model of acute ulcerative enteritis.

[0044] According to the following examples, it can be seen that the Faecalibacterium longum CM04-06 (CM04-06 was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on June 13, 2016, with a deposit number of CGMCC1.5208) selected by the present invention has a good effect in relieving ulcerative enteritis, which is specifically manifested in significantly relieving the apparent state of mice with ulcerative enteritis caused by DSS, and reducing the disease activity index and inflammatory response of mice.

[0045] The embodiments will be described in detail below.

[0046] Example 1 Screening and Identification of Faecalibacterium longum CM04-06

[0047] 1. Isolation and culture of CM04-06

[0048] The Faecalibacterium longum CM04-06 of the present invention is isolated from a fecal sample of a healthy child (male) in Shenzhen. The isolation and culture environment is strictly anaerobic conditions. The specific isolation process is: take about 0.2g of fecal sample in an anaerobic operation box, suspend and disperse it with 1mL of sterile PBS, shake and mix it thoroughly, and then dilute and apply it in a gradient. The culture medium uses anaerobic PYG culture medium. The specific components are shown in Table 1, where the composition of the inorganic salt solution is shown in Table 2:

[0049] Table 1: PYG medium formula

[0050] Components Content (1L) Peptone 5g Trypticase 5g Yeast powder 10g Beef extract 5g glucose 5g <![CDATA[K 2 HPO 4 ]]> 2g Sodium acetate 5g <![CDATA[Cysteine-HCl·H 2 The]]> 0.5g Heme 5mg Inorganic salt solution 40mL Resazurin 1mg Distilled water 950mL

[0051] Table 2: Inorganic salt solution formula

[0052]

[0053]

[0054] The coated plate was cultured at 37°C under anaerobic conditions for 3-4 days. After colonies grew on the surface of the plate, single colonies were picked for streaking and purification until pure culture strains were obtained. The isolated strains were frozen and stored at -80°C using glycerol.

[0055] 2. 16S rDNA identification of Faecalibacterium longum CM04-06

[0056] (1) Genome extraction: Cultivate the isolated strains until the bacterial concentration reaches 10 8 When the concentration is CFU / mL, take 2 mL of bacterial solution for genomic DNA extraction.

[0057] (2) PCR amplification of 16S rDNA: DNA was used as a template for 16S rDNA amplification. The universal amplification primers for 16S rDNA were 8F-1492R (5'-AGAGTTTGATCATGGCTCAG-3' and 5'-TAGGGTTACCTTGTTACGACTT-3'). The PCR amplification procedure was:

[0058]

[0059] (3) Purification and sequencing: The PCR product was purified by magnetic beads and then detected by electrophoresis. The band position of 16S rDNA was about 1.5k. The purified product was sequenced at 3730.

[0060] (4) Comparison of 16S rDNA sequence database: A 1372bp 16S rDNA sequence was obtained by sequencing. This sequence was compared with the EzTaxon-e database to obtain preliminary species classification information of the strain. The bacterium with the highest 16S rDNA homology to CM04-06 in the database is Faecalibacterium prausnitzii ATCC 27768 (hereinafter referred to as ATCC 27768), with a similarity of 96.87%. Based on the 16S rDNA information, it can be determined that CM04-06 is a new species belonging to the genus Faecalibacterium.

[0061] The 16S rDNA sequence of CM04-06 is:

[0062]

[0063] 3. 16S rDNA evolution analysis of CM04-06

[0064] The evolutionary analysis of CM04-06 was performed using 16S rDNA sequences. The 16S rDNA sequences of CM04-06 were compared with the EzBioCloud database to obtain species that were closely related to CM04-06. These species were compared with the 16S rDNA sequences of CM04-06, and then the neighbor-joining evolutionary tree was drawn using MegaX software ( Figure 1 ). According to the evolutionary tree, it can be further determined that CM04-06 belongs to the genus Faecalibacterium, and the species with the closest evolutionary relationship is ATCC 27768.

[0065] 4. Microbiological characteristics of CM04-06

[0066] (1) Morphological characteristics: CM04-06 was cultured at 37°C for 2-3 days in an anaerobic environment. The colonies were yellow-white, with a high water content, a little sticky, nearly round, opaque, flat with a convex center, and a colony diameter of about 2-3 mm.

[0067] (2) Microscopic characteristics: Under 1000x microscope magnification, the CM04-06 bacteria were long rod-shaped, Gram staining was negative, no spores or flagella were found, the diameter of the bacteria was about 1 μm, and the length was 4 to 10 μm.

[0068] (3) Physiological and biochemical characteristics: The oxidase and catalase reactions were negative, the growth temperature range was 30-45°C, the pH range was 4.0-9.0, and the optimum temperature and pH were 37°C and pH 7.0. It can tolerate 3% NaCl. The comparison of the physiological and biochemical reactions (including substrate utilization API 20A and enzyme reaction API ZYM) between CM04-06 and the closest related reference bacteria ATCC 27768 is shown in Table 3: (+, indicates positive reaction; -, indicates negative reaction; w indicates weak positive reaction).

[0069] Table 3: Biochemical characteristics of CM04-06 and ATCC 27768

[0070]

[0071] The above comparison of the physiological and biochemical reactions of CM04-06 with the reference bacteria showed that CM04-06 had significant differences in the utilization of raffinose and trehalose, hydrolysis of esculin and gelatin, and the activities of esterase (C4), valine arylaminease, cystine arylaminease, chymosin, naphthol-AS-BI-phosphohydrolase, β-uronidase, α-glucosidase and N-acetyl-glucosidase, which proved that CM04-06 was a new species different from the known bacteria.

[0072] 5. Analysis of Cellular Fatty Acids

[0073] The cells of CM04-06 and ATCC 27768 cultured to the stable phase were collected, and then the cell fatty acids were extracted and tested. The content and difference of the cell fatty acids of the two strains were analyzed by gas chromatography, and the specific results are shown in Table 4.

[0074] Table 4: Fatty acid content of CM04-06 and ATCC 27768 cells

[0075]

[0076] The analysis results of fatty acids showed that the main fatty acid composition (>10%) of CM04-06 was similar to that of the reference strain ATCC 27768, including 14-carbon saturated fatty acids, 16-carbon saturated fatty acids, and ω9c-18-carbon diunsaturated fatty acids, but the specific contents were somewhat different.

[0077] The above data on CM04-06 phenotype, 16S rDNA evolution analysis, physiological and biochemical reactions, and cytochemical composition showed that CM04-06 was a new species of the genus Faecalibacterium, and was named Faecalibacterium longum.

[0078] Example 2 Bioactive substances secreted by Faecalibacterium longum CM04-06

[0079] 1. Detection of short-chain fatty acids (SCFA)

[0080] (1) Sample preparation: Take 1 mL of CM04-06 bacterial culture solution that has been cultured for 48 hours, centrifuge at 12000 r / min for 5 minutes, and aspirate the supernatant for later use.

[0081] (2) Determination of SCFA: The determination of short-chain fatty acids was carried out by the external standard method, and acetic acid, propionic acid, butyric acid, and valeric acid were used to prepare the standard curve. Agilent gas chromatograph (GC-7890B, Agilent) was used, and HP-INNOWax (Cross-Linked PEG), 30m×0.25mm×0.25um capillary column was selected for analysis. The detector was a hydrogen flame ionization detector, and the GC parameters were set as column temperature: 180-200℃; vaporization chamber temperature: 240℃; detection temperature: 210℃; injection volume: 2μL; carrier gas flow rate: N 2 , 50mL / min; hydrogen flow rate: 50mL / min; air flow rate: 600~700mL / min.

[0082] (3) Results: The SCFA production was determined to be: formic acid (7.62 mmol / L), acetic acid (44.8 mmol / L), and butyric acid (40.03 mmol / L).

[0083] 2. Detection of organic acids

[0084] (1) Sample preparation: Same as above for SCFA detection

[0085] (2) Determination of organic acids: The standard substances for the detection of organic acids are: 3-methylbutyric acid, valeric acid, quinic acid, lactic acid, oxalic acid, malonic acid, benzoic acid, maleic acid, succinic acid, fumaric acid, malic acid, adipic acid, tartaric acid, shikimic acid, citric acid, isocitric acid and L-ascorbic acid. Agilent gas chromatograph (GC-7890B, Agilent) was used, and the chromatographic column was 122-5532G DB-5ms (40m×0.25mm×0.25um), column temperature: 270~290℃; injection port temperature: 250℃; gas flow rate: 0.86mL / min.

[0086] (3) Results: The organic acid yields obtained are shown in Table 5.

[0087] Table 5: Organic acid production of Faecalibacterium longum CM04-06

[0088] Organic Acid 3-Methylbutyric acid Valeric acid Quinic acid lactic acid oxalic acid Malonic acid Content (mmol / L) 0.26 0.49 0 30.53 0 0 Organic Acid benzoic acid Maleic acid Succinic acid Fumaric Acid Malic Acid Adipic acid Content (mmol / L) 1.79 0 0.88 0 0 0.60 Organic Acid tartaric acid Shikimic acid Citric Acid Isocitrate L-Ascorbic Acid Content (mmol / L) 0 0 0. 0 0.11

[0089] Example 3 Treatment of UC mice with Faecalibacterium longum CM04-06

[0090] The mouse model selected in this embodiment is: DSS-induced ulcerative enteritis mouse model, using C57bl / 6 mice (purchased from Guangdong Medical Experimental Animal Center), 8 weeks old, weighing 20g±2g, the mouse breeding environment is SPF grade, adaptive feeding for 1 week for DSS induction, and the mice are continuously fed with 2.5% DSS (molecular weight 36000-50000) for 7 days. VSL#3 (purchased from Sigma Tau, USA) was used as a positive control to compare the therapeutic effects of Faecalibacterium longum CM04-06 and VSL#3.

[0091] A total of 48 experimental mice were randomly divided into 4 groups, with 15 mice in each group, including a control group, a DSS-induced model group (each mouse was gavaged with 0.2 ml PBS per day), a Faecalibacterium CM04-06 treatment group, and a VSL#3 treatment group.

[0092] The treatment process of Faecalibacterium longitubaum CM04-06 was as follows: the Faecalibacterium longitubaum CM04-06 bacterial solution was cultured anaerobically for 24 hours, the cells were collected by centrifugation, suspended with PBS, and the bacterial concentration was adjusted to 1×10 9 CFU / mL, each mouse was gavaged with 200 μL of Faecalibacterium CM04-06 every day.

[0093] The VSL#3 treatment process is as follows: PBS was used to suspend the cells and the concentration was adjusted to 1×10 9 CFU / mL, each mouse was gavaged with 200 μL per day.

[0094] The treatment was carried out in the form of preventive administration. Probiotics were gavaged three days before DSS treatment. The weight, diet and drinking water of the mice were recorded every day. The fecal characteristics and fecal occult blood of the mice were observed. The DAI scores of the mice were calculated on the 1st, 3rd, 5th and 7th days of DSS treatment. The DAI scores are shown in Table 6. DSS treatment lasted for 7 days, and the daily gavage volume of probiotics and PBS was 200 μL. After the experiment, the mice were killed, and all mice were bled, decapitated, colons were removed, photographed, weighed, and colon lengths were measured. Colon tissues were stored in a -80°C refrigerator and in paraformaldehyde.

[0095] Table 6: DAI Index Score Table

[0096] Weight loss (%) Stool characteristics Occult blood in stool / bleeding blood in stool integral 0 normal normal 0 1-5 1 5-10 Loose Occult blood positive 2 10-15 3 >15 Loose stools Naked bloody stool 4

[0097] The stool characteristics in the table are: normal stool - formed stool; loose stool - pasty, semi-formed stool that does not adhere to the anus; loose stool - watery stool that can adhere to the anus. Among them, blood in the stool: normal mice are positive for blood in the stool; blood in the stool that can be seen is red or brown; occult blood positive is not obvious blood in the stool, which is detected using tetramethylbenzidine. The DAI index is equal to the sum of the three scores of body weight, stool characteristics, and occult blood / weak blood in the stool.

[0098] Test results:

[0099] (1) The weight changes of mice before and after treatment are shown in Table 7 and Figure 2 As shown:

[0100] Table 7: Effects of CM04-06 on body weight of mice

[0101]

[0102]

[0103] From Table 7 and Figure 2 It can be seen that the weight of mice in the control group basically maintained a slow increase trend, and the weight of mice in the DSS-induced model group gradually decreased. The weight loss was more significant from the 3rd day (*P<0.05), and the difference between the two was more obvious from the 5th day (**P<0.01). The treatment of CM04-06 and VSL#3 can slow down the weight loss of UC mice. On the 5th and 7th days, the inhibition of weight loss of CM04-06 and VSL#3 mice was significantly different from that of the model group (▲P<0.05). The above results show that the two groups of probiotics, CM04-06 and VSL#3, can inhibit the weight loss of mice caused by UC. By comparing the weight values ​​of each group on the 7th day, it can be found that the weight of mice in the CM04-06 group was slightly higher than that of VSL#3, and the inhibition of weight loss of mice was more significant (▲▲P<0.01), indicating that CM04-06 is better than VSL#3 in inhibiting the weight loss of UC mice.

[0104] (2) Changes in DAI scores of mice before and after treatment

[0105] The DAI index of mice with DSS-induced ulcerative enteritis changed due to weight loss, stool characteristics and blood in stool. The specific results are shown in Table 8 and Figure 3 :

[0106] Table 8: Effects of CM04-06 on DAI scores of mice

[0107] Grouping Day 1 Day 3 Day 5 Day 7 Control group 1.0±0.5 1.2±0.6 1.1±0.6 1.3±0.5 Model Group 1.2±0.4 3.5±1.1** 6.8±1.6** 8.9±1.9** CM04-06 1.1±0.5 3.1±1.2 <![CDATA[5.2±1.5 ▲▲ ]]> <![CDATA[7.0±1.8 ▲▲ ]]> <![CDATA[VSL # 3]]> 1.2±0.6 3.3±1.1 <![CDATA[5.8±1.8 ▲▲ ]]> <![CDATA[7.4±1.6 ▲ ]]>

[0108] Table 8 and Figure 3The data showed that the DAI scores of mice in the control group were maintained at a stable low level, while the DAI scores of mice induced by DSS gradually increased due to a series of pathologies. Starting from the third day, the DAI scores of mice in the model group were significantly different from those in the control group (**P<0.01), and the DAI scores of mice in the model group reached the highest level on the seventh day. The intervention of probiotics can inhibit the increase of DAI scores. The DAI scores of mice in the CM04-06 and VSL#3 groups on the fifth and seventh days were controlled to a certain extent compared with the model group (▲P<0.05, ▲▲P<0.01). The results on the seventh day showed that the DAI scores of mice in the CM04-06 group were slightly lower than those in the VSL#3 group, and the DAI scores of the CM04-06 group and the model group were significantly different (▲▲P<0.01), which shows that CM04-06 is better than VSL#3 in inhibiting the increase of DAI scores in UC mice.

[0109] (3) Changes in colon length

[0110] The colon tissue of UC model mice will change, mainly because the occurrence of ulcers and inflammation leads to shortening of the colon tissue. After the treatment, the colon length of the mice was dissected and measured. The experimental results are shown in Table 9.

[0111] Table 9: Effects of CM04-06 on colon length in mice

[0112] Grouping Colon length (cm) Control group 7.16±0.82 Model Group 4.58±0.98** <![CDATA[VSL # 3 groups]]> <![CDATA[5.43±0.90 ▲ ]]> CM04-06 Group <![CDATA[5.70±0.85 ▲ ]]>

[0113] The results in Table 9 show that the shortening of the colon tissue of mice (model group) induced by DSS for 7 days was more serious, and the difference was extremely significant compared with the control group (**P<0.01). The intervention of probiotics CM04-06 and VSL#3 can significantly inhibit the shortening of the mouse colon, and the differences between CM04-06 and VSL#3 and the model group were significant (▲P<0.05). From the data in Table 9, it can be found that the colon length of mice in the CM04-06 group is longer than that of mice in the VSL#3 group, indicating that CM04-06 is slightly better than VSL#3 in controlling the shortening of the colon of UC mice.

[0114] Example 4 Preparation of capsules containing Faecalibacterium longum CM04-06

[0115] Table 10: Capsule raw material formula

[0116]

[0117]

[0118] Lactose, yeast powder, and peptone were mixed evenly with purified water according to the formula ratio described in Table 10, preheated to 60-65°C, homogenized at 20 MPa pressure, sterilized at about 90°C for 20-30 minutes, cooled to 36-38°C, mixed with protective agent vitamin C, and inoculated with live Faecalibacterium CM04-06 bacteria (1-500×10 6 CFU / mL), fermented at 36-38°C to a pH of 6.0, centrifuged, and freeze-dried until the moisture content is less than 3%, thereby preparing a freeze-dried product of Faecalibacterium longiflorum CM04-06. Weigh 0.5 g of the freeze-dried product of Faecalibacterium longiflorum CM04-06 and mix it with maltodextrin in equal amounts and then put it into a capsule to prepare a pharmaceutical composition containing Faecalibacterium longiflorum CM04-06.

[0119] Example 5 Preparation of a preparation for treating ulcerative enteritis

[0120] 1. Preparation of bacterial solution: Faecalibacterium longum CM04-06 (1×10 9 CFU / mL) was used for anaerobic culture, and the anaerobic culture medium used PYG culture medium, which was fermented anaerobicly at 37°C for 2 to 3 days.

[0121] 2. Preparation of growth factors: skim milk and casein are mixed, centrifuged and ultrafiltered to obtain a crude extract of milk growth factors (containing vitamins, purines, pyrimidines and other ingredients).

[0122] 3. Preparation of drug dosage form: Add 5 volumes of growth factor and 1 volume of protective agent vitamin C to 100 volumes of CM04-06 fermented bacterial solution, stir well to mix, and then add starch auxiliary material (such as maltodextrin) to prepare drug dosage form.

[0123] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0124] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. Use of Faecalibacterium longum in preparing a composition for treating enteritis, wherein the Faecalibacterium longum was deposited in the General Microbiological Center of China Microbiological Culture Collection Administration on June 13, 2016, with a deposit number of CGMCC 1.5208, and the composition is used for at least one of the following: Inhibits enteritis-induced weight loss; Suppress the increase of disease activity index caused by enteritis; and Improve intestinal changes caused by enteritis, in, The enteritis is ulcerative enteritis; the intestinal changes are shortening of colon tissue.

2. The use according to claim 1, characterized in that The ulcerative enteritis is ulcerative colitis.

3. The use according to claim 1, characterized in that The enteritis was caused by DSS.

4. The use according to claim 1, characterized in that The concentration of the long-lived Faecalibacterium was 1.0×10 6 CFU / mL~1.0×10 9 CFU / mL.

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