CN114231470A - Lactobacillus acidophilus capable of relieving ulcerative colitis and application thereof - Google Patents
Lactobacillus acidophilus capable of relieving ulcerative colitis and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses lactobacillus acidophilus capable of relieving ulcerative colitis and application thereof, and belongs to the technical field of microorganisms. The invention screens out a lactobacillus acidophilus CCFM1200, and the lactobacillus acidophilus CCFM1200 has the function of relieving ulcerative colitis caused by DSS, and is specifically represented by the following steps: (1) reducing weight loss in ulcerative colitis mice; (2) the colon length of an ulcerative colitis mouse is obviously increased; (3) the level of immune factors in the colon of the ulcerative colitis mouse is obviously improved; (4) the expression level of a cell chemotactic factor receptor in the colon of an ulcerative colitis mouse is obviously reduced; (5) the content of short-chain fatty acid in the excrement of the ulcerative colitis mouse is obviously improved; (6) regulating intestinal flora of mice with ulcerative colitis. The lactobacillus acidophilus CCFM1200 has a huge application prospect in the aspect of preparing foods or medicines for preventing and/or treating ulcerative colitis.
Description
Technical Field
The invention relates to lactobacillus acidophilus capable of relieving ulcerative colitis and application thereof, in particular to a strain capable of relieving ulcerative colitis, which can be added into various health foods and belongs to the technical field of microorganisms.
Background
Inflammatory Bowel Disease (IBD) mainly includes Crohn's Disease (CD) and ulcerative colitis, a chronic inflammation of the intestinal tract characterized by an impaired intestinal epithelial barrier function and a dysregulated intestinal immunity. When the integrity of the intestinal epithelium is compromised, bacteria and their products in the gut can penetrate the intestinal barrier, resulting in abnormal immune responses and inflammation. In recent years, the number of people suffering from IBD in China is on a rapid rise. Currently, the etiology of inflammatory bowel disease is not well defined. Therapeutic drugs commonly used in inflammatory bowel disease are: aminosalicylic acids, hormones and immunosuppressants. However, the three drug treatment modes have more side effects and more limitations for treating the inflammatory bowel disease.
In recent years, the use of probiotics for the alleviation of inflammatory bowel disease by probiotics has become a focus of research. Many studies have shown that many probiotic strains have some degree of relief from inflammatory bowel disease. The probiotics can be colonized in a human body, the composition of host intestinal flora is changed, and beneficial metabolites are produced at the same time, so that beneficial effects are produced on the host. Compared with common medicines, the probiotics has the advantages of high safety, no side effect, no drug resistance, low cost and the like. Therefore, it is urgently needed to find a probiotic strain capable of relieving ulcerative colitis.
Disclosure of Invention
The invention provides Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200 which is classified and named as Lactobacillus acidophilus and is preserved in Guangdong province microorganism strain preservation center in 12 and 15 months 2021, wherein the preservation number is GDMCC No. 62128, and the preservation address is No. 59 building 5 of Mieli Zhonglu 100, Guangzhou city.
In one embodiment, the lactobacillus acidophilus CCFM1200 has the 16S rDNA sequence shown in SEQ ID No.1, and the colonies on MRS medium appear white, rough on the surface, and dull.
The invention also provides a probiotic agent containing the lactobacillus acidophilus CCFM 1200.
In one embodiment, the probiotic comprises lactobacillus acidophilus CCFM1200 with a viable count not less than 1 × 109CFU/mL or 1X 109CFU/g。
In one embodiment, the probiotic is a suspension of lactobacillus acidophilus CCFM 1200.
The invention also provides a starter containing the lactobacillus acidophilus CCFM 1200.
In one embodiment, the preparation method of the leavening agent is as follows: inoculating lactobacillus acidophilus CCFM1200 into a culture medium, and culturing at 37 ℃ for 24-48 h to obtain a culture solution; centrifuging the culture solution to obtain thalli; the cells were resuspended in physiological saline to obtain a starter culture.
In one embodiment, the medium is MRS medium.
The invention also provides a medicament containing the lactobacillus acidophilus CCFM 1200.
In one embodiment, the number of viable bacteria of Lactobacillus acidophilus CCFM1200 in said medicament is not less than 1 × 109CFU/mL or 1X 109CFU/g。
In one embodiment, the medicament comprises said lactobacillus acidophilus CCFM1200 and a pharmaceutically acceptable carrier.
The invention also provides application of the lactobacillus acidophilus CCFM1200 in preparing a medicament for preventing and/or treating ulcerative colitis.
In one embodiment, the prevention and/or treatment of ulcerative colitis comprises at least one of the following effects:
(1) alleviating weight loss due to ulcerative colitis;
(2) improving colon length reduction due to ulcerative colitis;
(3) increasing immune factor IL-10 levels in the colon;
(4) reducing the mRNA expression level of cell chemotactic factor receptors CCR2 and CCR1 in colon.
In one embodiment, the medicament is also for increasing the content of short chain fatty acids in the intestinal tract.
In one embodiment, the medicament is also for modulating the intestinal flora.
In one embodiment, said modulating gut flora comprises decreasing the relative abundance of proteobacteria in the gut flora.
In one embodiment, the modulating gut flora comprises decreasing the relative abundance of escherichia _ Shigella in the gut flora.
Has the advantages that:
1. the invention screens out a Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200, and the Lactobacillus acidophilus CCFM1200 has the function of relieving ulcerative colitis caused by dextran sodium sulfate and is specifically embodied in that:
(1) reducing weight loss in ulcerative colitis mice;
(2) the colon length of an ulcerative colitis mouse is obviously improved;
(3) the level of immune factors in the colon of the ulcerative colitis mouse is obviously improved;
(4) the content of short-chain fatty acid in the excrement of the ulcerative colitis mouse is obviously improved;
(5) the diversity of intestinal flora of the ulcerative colitis mouse is obviously improved;
(6) reducing the relative abundance of proteobacteria in intestinal flora of ulcerative colitis mice;
(7) reducing the relative abundance of Escherichia _ Shigella in intestinal flora of mice with ulcerative colitis;
(8) significantly reducing the expression level of mRNA of a cell chemotactic factor receptor in the colon of an ulcerative colitis mouse;
based on the relieving and treating application of lactobacillus acidophilus CCFM1200 to ulcerative colitis mice, the lactobacillus acidophilus CCFM1200 of the invention has huge application prospect in preparing products (such as food or medicine) for preventing and/or treating ulcerative colitis.
2. The culture process of the lactobacillus acidophilus only needs to control a culture medium and some culture conditions, the cost is relatively low, and the industrial production is easy to realize.
Biological material preservation
Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200 is classified and named Lactobacillus acidophilus, is preserved in Guangdong province microorganism strain preservation center 12 and 15 days 2021, and has the preservation number of GDMCC No. 62128, and the preservation address of No. 59 building 5 of Ji-Hai-100 of Jieli Zhou-Luo, Guangzhou city.
Drawings
FIG. 1: body weight change in different groups of mice.
FIG. 2: colon length status in different groups of mice.
FIG. 3: the IL-10 content in the colon of different groups of mice.
FIG. 4: the content of acetic acid in the feces of different groups of mice.
FIG. 5: the content of butyric acid in the feces of different groups of mice.
FIG. 6: the alpha diversity of the intestinal flora of different groups of mice is Chao1 index.
FIG. 7: the alpha diversity Shannon index of the intestinal flora of different groups of mice.
FIG. 8: relative abundance of the enteromorpha proteobacteria of different groups of mice.
FIG. 9: relative abundance of intestinal flora escherichia _ Shigella in different groups of mice.
FIG. 10: relative expression of CCR2 mRNA in colon of different groups of mice.
FIG. 11: relative expression of CCR1 mRNA in colon of different groups of mice.
Detailed Description
The invention is further elucidated with reference to a specific embodiment and a drawing.
C57BL/6J mice referred to in the examples below were purchased from Viton, Zhejiang; dextran sulfate sodium referred to in the examples below was purchased from MP Biomedicals; lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200 referred to in the following examples was isolated from the Biotechnology center of the food institute of south Jiangnan university; lactobacillus acidophilus NCFM was obtained from dupont china group ltd; studies have demonstrated that lactobacillus acidophilus strain NCFM is a type of probiotic that is present in healthy humans (mainly in the human gut environment). Since the lactobacillus acidophilus strain NCFM is separated, extensive research is carried out, and the lactobacillus acidophilus strain NCFM has the effects of adjusting and improving gastrointestinal tract functions, improving the immunity of organisms, relieving ulcerative colitis and the like. Lactobacillus acidophilus NCFM has become a commercial strain and has been reported to have beneficial effects in alleviating ulcerative colitis caused by sodium dextran sulfate. Therefore, in the following examples, Lactobacillus acidophilus NCFM was used as a positive control for the strain.
The detection reagents referred to in the following examples are as follows:
ELISA kits for IL-10 (cat # DY417) and IL-17 (cat # DY421) were purchased from R & D Systems.
The media involved in the following examples are as follows:
MRS solid medium: 10g/L of peptone, 10g/L of beef extract, 20g/L of glucose, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate2PO4·3H2O 2.6g/L、MgSO4·7H2O 0.1g/L、MnSO40.05g/L, Tween 801 mL/L and agar 15 g/L.
MRS liquid medium: 10g/L of peptone, 10g/L of beef extract, 20g/L of glucose, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate2PO4·3H2O 2.6g/L、MgSO4·7H2O 0.1g/L、MnSO40.05g/L and Tween 801 mL/L.
Example 1: screening and strain identification of lactobacillus acidophilus CCFM1200
1. Screening
Taking a healthy human body excrement sample from Wen Wuhu City of Anhui province, taking 0.5g of the sample stored in 30% (v/v) glycerol, and adding the sample into a 10mL centrifuge tube filled with 4.5mL of normal saline under a sterile environment to obtain 10-1Diluting the solution, repeating the above dilution steps to obtain 10-2、10-3、10-4、10-5、10-6Diluting the solution; respectively sucking 100 mu L of gradient dilution liquid with different gradients, coating the gradient dilution liquid on an MRS solid culture medium, and culturing for 72h at 37 ℃ to obtain a diluted coating plate; selecting typical colonies on the diluted coating plate, streaking on an MRS solid culture medium, and culturing at 37 ℃ for 48h to obtain purified colonies; and (3) selecting a purified colony, inoculating the colony into an MRS liquid culture medium, and culturing at 37 ℃ for 48h to obtain the strain CCFM 1200.
2. Identification
The genome of CCFM1200 was extracted, the 16S rDNA of CCFM1200 was amplified and sequenced (by Jinzhi Biotech, Suzhou), and the 16S rDNA sequence of CCFM1200 (the 16S rDNA sequence of CCFM1200 is shown in SEQ ID NO. 1) obtained by sequencing analysis was aligned in GenBank, and the strain was Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM 1200.
Example 2: preparation of Lactobacillus acidophilus suspension
The preparation method of the lactobacillus acidophilus NCFM or lactobacillus acidophilus CCFM1200 bacterial liquid comprises the following steps:
streaking lactobacillus acidophilus liquid on an MRS solid culture medium, and culturing at 37 ℃ for 48h to obtain a single colony;
selecting a single colony, inoculating the single colony into an MRS liquid culture medium, and culturing for 24h at 37 ℃ to obtain an activating solution; inoculating the activated solution into an MRS liquid culture medium according to the inoculation amount of 1% (v/v), and culturing at 37 ℃ for 24h to obtain a first-level seed solution;
inoculating the primary seed liquid into an MRS liquid culture medium according to the inoculation amount of 1% (v/v), and culturing at 37 ℃ for 24h to obtain a secondary seed liquid;
inoculating the secondary seed liquid into an MRS liquid culture medium according to the inoculation amount of 1% (v/v), and culturing at 37 ℃ for 24h to obtain a bacterial liquid; centrifuging 6000g of the bacterial liquid for 15min, and collecting precipitates; washing the precipitate with physiological saline buffer solution twice, and centrifuging again at 6000g for 10min to obtain thallus; resuspending the lactobacillus cells in physiological saline to a cell concentration of 1X 109CFU/mL to obtain Lactobacillus acidophilus liquid.
Example 3: effect of Lactobacillus acidophilus CCFM1200 on body weight and colon Length in ulcerative colitis mice
Taking 32 male pathogen free (SPF) C57BL/6J mice 6-8 weeks old, breeding the mice for 1 week under the conditions of room temperature of 22-24 ℃, humidity of 40-60%, day and night alternation of 12h/12h, free eating and drinking, and then randomly dividing the mice into 4 groups, wherein each group comprises 8 mice, and the 4 groups comprise: a control group, a modeling group, a Lactobacillus acidophilus (Lactobacillus acidophilus) NCFM group and a Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200 group.
The experiment was started 1 week after the animals were acclimatized for a total of 2 weeks. The specific treatment is as follows:
control group: mice normally drink water throughout the experimental period and are gavaged with 200 μ L of physiological saline as a control;
building a module: normally drinking water on days 1-7, drinking 2.5% (w/v) dextran sodium sulfate solution from day 8, continuing for 7 days after molding, and perfusing 200 mu L normal saline in the whole experiment process;
lactobacillus acidophilus NCFM group: normally drinking water on day 1-7, drinking 2.5% (w/v) dextran sodium sulfate solution from day 8, continuing for 7 days after molding, and perfusing mouse with 200 μ L bacterial suspension every day in the whole experiment process to make the perfusateThe amount is 1X 109CFU/only/day.
Lactobacillus acidophilus CCFM1200 group: normally drinking water on day 1-7, drinking 2.5% (w/v) dextran sodium sulfate solution from day 8, continuing for 7 days after molding, and intragastrically administering 200 μ L bacterial suspension to mice every day in the whole experiment process to make intragastrically administered dose be 1 × 109CFU/only/day.
Measuring the weight of each group of mice by a weighing machine during the molding and after the molding is finished; after the experiment is finished, taking the colon of the mouse, measuring the length of the colon of the mouse, and respectively showing the measurement results in figures 1-2.
As can be seen from fig. 1, the body weight ratios (body weight ratio on day 7 to day 1) of the control, model, NCFM, and CCFM1200 mice were 1.02, 0.91, 0.93, and 0.94, respectively, after the model was made up to day 7. The NCFM group and CCFM1200 group reduced the weight loss of mice when molded with dextran sodium sulfate.
As can be seen from fig. 2, the colon length of the control and CCFM1200 mice was significantly higher than that of the model mice (122% and 115%, respectively), but the colon length of the NCFM mice was not significantly different from that of the model mice. Therefore, the Lactobacillus acidophilus CCFM1200 can relieve the weight reduction of the ulcerative colitis mouse, obviously improve the symptom of colon length reduction, and has better effect than Lactobacillus acidophilus NCFM.
Example 4: effect of Lactobacillus acidophilus CCFM1200 on immune factor levels in Colon of ulcerative colitis mice
The mice were grouped and modeled as in example 3. After the experiment is finished, blood is taken and mice are killed, the colons of the mice are taken and placed in phosphate buffer saline solution and homogenized, the content of IL-10 in the colons of each group of mice is measured by an ELISA kit, and the detection result is shown in figure 3.
As shown in FIG. 3, the concentration of IL-10 in the colon of the control mice was 26.22pg/mg protein, and the IL-10 level of the model group was decreased to 14.05pg/mg protein compared to the control group. Compared with the model-making group mice, the level of IL-10 in the colon of the NCFM group and the CCFM1200 group is obviously improved, namely 36.18pg/mg protein and 44.55pg/mg protein respectively, and the effect of the CCFM1200 group is obviously better than that of the NCFM group.
Therefore, the Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200 can improve the level of the immune factor IL-10 in the colon of the ulcerative colitis mouse, and the effect is better than that of the Lactobacillus acidophilus (Lactobacillus acidophilus) NCFM.
Example 5: effect of Lactobacillus acidophilus CCFM1200 on short-chain fatty acid content in feces of mice with ulcerative colitis
The mice were grouped and modeled as in example 3. After the experiment is finished, collecting the mouse excrement, placing the mouse excrement in liquid nitrogen, transferring the liquid nitrogen to a refrigerator at the temperature of-80 ℃, taking out the mouse excrement before detecting the content of the short-chain fatty acid, carrying out vacuum freeze drying, accurately weighing 0.05g of the freeze-dried excrement sample, dissolving the freeze-dried excrement sample in 0.5mL of saturated sodium chloride solution, soaking for 30min, homogenizing the tissue homogenizer, adding 0.02mL of sulfuric acid with the concentration of 10%, shaking for 30s, accurately adding 0.8mL of ether solution into the excrement solution in a ventilation cabinet, shaking for 30s, centrifuging for 15min (8000g and 4 ℃), transferring supernatant into a centrifugal tube containing 0.25g of anhydrous sodium sulfate, shaking uniformly, centrifuging for 15min (8000g and 4 ℃), taking the supernatant into a gas volumetric flask, detecting the content of the short-chain fatty acid through GCMS, and detecting results are shown in figures 4 and 5.
As shown in FIGS. 4 and 5, the contents of acetic acid and butyric acid (46.77. mu. mol/g, 37.38. mu. mol/g, respectively) in the feces of the mice in the molding group are reduced compared with those of the mice in the control group (54.69. mu. mol/g, 46.04. mu. mol/g, respectively); the content of acetic acid and butyric acid in the feces of mice in the CCFM1200 group (77.74 mu mol/g, 59.15 mu mol/g) is obviously up-regulated compared with that of the model group; the contents of acetic acid and butyric acid in the feces of the mice in the NCFM group are not obviously different from those of the model group.
Therefore, the content of acetic acid and butyric acid in the excrement of the mice with ulcerative colitis can be obviously improved by Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200, and the effect is not achieved by Lactobacillus acidophilus (Lactobacillus acidophilus) NCFM.
Example 6: influence of lactobacillus acidophilus CCFM1200 on diversity of intestinal flora of ulcerative colitis mice
The mice were grouped and modeled as in example 3. After completion of the experiment, mouse feces were collected, genomic DNA in the feces was extracted with the FastDNA Spin Kit (MP Bio-pharmaceuticals, USA), and V3-V4 as genomic DNA was extracted
The regions were subjected to specific PCR amplification, 16S rDNA sequencing, and the fecal flora alpha diversity (Chao 1 and Shannon) was analyzed, and the results are shown in FIGS. 6 and 7.
As can be seen from fig. 6, in terms of the Chao1 index, Lactobacillus acidophilus (21.5) NCFM (47.25) and Lactobacillus acidophilus (50.875) did not significantly differ between the control group (21.5) and the model group (24.38) and significantly increased the diversity of the mouse intestinal flora.
As can be seen from fig. 7, in terms of Shannon index, Lactobacillus acidophilus (Lactobacillus acidophilus) NCFM (2.84), Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200(2.80) also significantly improved the diversity of the mouse intestinal flora relative to the control group (2.25) and the model group (2.52).
Example 7: effect of Lactobacillus acidophilus CCFM1200 on Enterobacteriaceae Proteobacteria of ulcerative colitis mice
The mice were grouped and modeled as in example 3. After the experiment is finished, collecting mouse feces, extracting genomic DNA in the feces by using a FastDNA Spin Kit (MP biological medicine company, USA), carrying out specific PCR amplification on a V3-V4 region of the extracted genomic DNA, carrying out 16S rDNA sequencing, analyzing the change of proteobacteria in the fecal flora, and obtaining an analysis result shown in figure 8.
As can be seen from FIG. 8, the abundance of Proteobacteria (4.65%) in the feces of the mice in the molding group was significantly increased compared to the control group (0.52%). Lactobacillus acidophilus (Lactobacillus acidophilus) NCFM (2.61%), Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200 (2.50%) were able to down-regulate the relative abundance of proteobacteria in mouse feces by 2.04% and 2.15%, respectively.
Example 8: effect of Lactobacillus acidophilus CCFM1200 on intestinal Escherichia _ Shigella in mice with ulcerative colitis
The mice were grouped and modeled as in example 3. After completion of the experiment, mouse feces were collected, genomic DNA in the feces was extracted with the FastDNA Spin Kit (MP Bio-pharmaceuticals, USA), and V3-V4 as genomic DNA was extracted
The region was subjected to specific PCR amplification, 16S rDNA sequencing, and the change in Escherichia _ Shigella in fecal flora was analyzed, and the results of the analysis are shown in FIG. 9.
As can be seen from FIG. 9, the relative abundance of Escherichia _ Shigella in the feces of the model-made mice (3.12%) was increased by 2.74% relative to the control (0.38%). Lactobacillus acidophilus (Lactobacillus acidophilus) NCFM (0.34%), Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200 (0.38%) were able to down-regulate the relative abundance of Escherichia _ Shigella in mouse feces by 2.78% and 2.74%, respectively.
Example 9: effect of Lactobacillus acidophilus CCFM1200 on the expression levels of intestinal CCR2 and CCR1 in ulcerative colitis mice
The mice were grouped and modeled as in example 3. After the experiment is finished, blood is taken and mice are killed, colon of the mice is taken to extract RNA, reverse transcription is carried out by using the kit, the expression levels of the cell chemotactic factors CCR2 and CCR1 in the colon of each group of mice are measured, and the detection result is shown in a figure of 10-11.
As shown in fig. 10, the expression level of CCR2 in colon sites of mice after DSS molding was significantly increased compared to the blank group of mice. After being treated by lactobacillus acidophilus NCFM and lactobacillus acidophilus CCFM1200, the relative expression amount is respectively and obviously reduced by 71.67 percent and 84.41 percent compared with that of the model building component.
As shown in fig. 11, the expression level of CCR1 in colon sites of mice after DSS molding was significantly increased compared to the blank group of mice. After being treated by lactobacillus acidophilus NCFM, the relative expression quantity is reduced by 44.21 percent compared with that of the model building group. After being treated by lactobacillus acidophilus CCFM1200, the relative expression quantity of the lactobacillus acidophilus CCFM is obviously reduced by 54.83 percent compared with that of a model group.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
SEQUENCE LISTING
<110> university of south of the Yangtze river
<120> lactobacillus acidophilus capable of relieving ulcerative colitis and application thereof
<130> BAA211620A
<160> 1
<170> PatentIn version 3.3
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<211> 1430
<212> DNA
<213> Lactobacillus acidophilus
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catcggaaac tgtttttctt gagtgcagaa gaggagagtg gaactccatg tgtagcggtg 660
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cgctgaggct cgaaagcatg ggtagcgaac aggattagat accctggtag tccatgccgt 780
aaacgatgag tgctaagtgt tgggaggttt ccgcctctca gtgctgcagc taacgcatta 840
agcactccgc ctggggagta cgaccgcaag gttgaaactc aaaggaattg acgggggccc 900
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Claims (10)
1. Lactobacillus acidophilus (Lactobacillus acidophilus) CCFM1200, which has been deposited in Guangdong province collection center for microorganism strains in 12 months and 15 days in 2021 with the deposit number being GDMCC No. 62128.
2. A probiotic comprising lactobacillus acidophilus CCFM1200 according to claim 1.
3. The probiotic preparation according to claim 2, characterized in that the viable count of Lactobacillus acidophilus CCFM1200 in the probiotic is not less than 1 x 109CFU/mL or 1X 109CFU/g。
4. Food product comprising lactobacillus acidophilus CCFM1200 according to claim 1.
5. A pharmaceutical composition comprising the Lactobacillus acidophilus CCFM1200 of claim 1.
6. The medicament of claim 5, comprising Lactobacillus acidophilus CCFM1200 of claim 1 and a pharmaceutically acceptable carrier.
7. The medicament according to claim 5 or 6, which isIs characterized in that the viable count of the lactobacillus acidophilus CCFM1200 is not less than 1 multiplied by 109CFU/mL or 1X 109CFU/g。
8. Use of lactobacillus acidophilus CCFM1200 according to claim 1 for the preparation of a medicament for the prevention and/or treatment of ulcerative colitis.
9. Use according to claim 8, wherein the prevention and/or treatment of ulcerative colitis comprises at least one of the following effects:
(1) alleviating weight loss due to ulcerative colitis;
(2) improving colon length reduction due to ulcerative colitis;
(3) increasing immune factor IL-10 levels in the colon;
(4) reducing the mRNA expression level of cell chemotactic factor receptors CCR2 and CCR1 in colon.
10. The use according to claim 8, wherein the medicament is also for increasing the content of short chain fatty acids in the intestinal tract.
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