A composition with digestive tract protection and repairing effects and its application

Abstract

The invention discloses a composition with digestive tract protection and repairing effect and its application, and belongs to the field of microbial engineering technology. The raw materials made of it include Lactobacillus paracasei N1115, 2'-fucosyllactose and lactose-N-neotetraose with a weight ratio of 1: 10000 ~ 15000: 3000 ~ 5000. A composition with digestive tract protection and repairing effect provided by the present invention, Lactobacillus paracasei N1115, 2'-FL and LNnT have a synergistic effect in anti-DSS-induced inflammatory response and repairing inflammatory damage, and the optimal content of Lactobacillus paracasei N1115 is also verified by a large number of experiments, and the excessively high concentration of bacteria content will reduce the effect. The present invention is suitable for preparing dairy products with digestive tract protection and repairing effect.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial engineering, and relates to a composition comprising Lactobacillus paracasei N1115, in particular to a composition with digestive tract protection and repairing effects and an application thereof. Background Art

[0002] Digestive tract damage is a common and recurrent digestive tract disease. Some drugs, such as nonsteroidal anti-inflammatory drugs (NSAIDs), can cause digestive tract damage by irritating the gastrointestinal mucosa, which can cause abdominal pain and diarrhea in mild cases, and can cause digestive system, circulatory system, immune system and other diseases in severe cases, causing patients to lose weight and reduce their quality of life.

[0003] In the prior art, non-biological drugs are usually used to alleviate or repair digestive tract damage, but such drugs have significant side effects and have an impact on the health of the body. Other biological drugs or health products, such as probiotics, have very different effects in terms of digestive tract protection and repair, and their quality varies greatly. Summary of the invention

[0004] The purpose of the present invention is to provide a composition having digestive tract protection and repair effects and its application, so as to provide a product with better digestive tract protection and repair effects.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is: A composition with digestive tract protection and repairing effects. The raw materials for making the composition include Lactobacillus paracasei N1115, 2'-fucosyllactose (2'-FL) and lactose-N-neotetraose (LNnT) in a weight ratio of 1:10000-15000:3000-5000.

[0006] As a limitation, the viable bacterial count of the Lactobacillus paracasei N1115 is 5×10 9 ~9×10 11 CFU / g.

[0007] The present invention also provides the application of the above composition, which is used for preparing probiotic products or dairy products with digestive tract protection and repair effects.

[0008] The present invention also provides a dairy product with digestive tract protection and repairing effects. The dairy product is prepared by adding the above composition into whole milk powder.

[0009] As a limitation, the addition amount of the composition is 6.5~15kg / t.

[0010] Due to the adoption of the above technical solution, the present invention has the following technical advances compared with the prior art: The present invention provides a composition with digestive tract protection and repair effects. Lactobacillus paracasei N1115, 2'-FL and LNnT have synergistic effects in resisting DSS (dextran sulfate sodium, commonly used in mouse colitis modeling)-induced inflammatory response and repairing inflammatory damage. The optimal content of Lactobacillus paracasei N1115 has also been verified by a large number of experiments. Too high a concentration of bacteria will reduce the effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 This is a graph showing the results of the mouse weight test in Example 1; Figure 2 This is a graph showing the DAI scoring results of mice in Example 1; Figure 3 This is a colonoscopy photo of mice in Example 1. DETAILED DESCRIPTION

[0012] The present invention is further described in detail below by specific examples. It should be understood that the described examples are only used to explain the present invention, and are not intended to limit the present invention.

[0013] Unless otherwise specified, the experimental methods used in the following examples are conventional methods in the art.

[0014] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0015] In the following embodiments: Lactobacillus paracasei N1115 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on March 17, 2011, with the deposit number of CGMCC No.4691 and the Latin name of Lacticaseibacillus paracasei , first disclosed in the Chinese invention patent with application number “201110357058.8” and titled “Lactobacillus casei N1115, its immunomodulatory effect and application”; Lactobacillus paracasei JMCC0012, the strain was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on November 23, 2015, with the deposit number CGMCC No.11726, and the Latin name is Lacticaseibacillus paracasei ; The storage address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. On August 25, 2022, the Department of Food Safety Standards, Monitoring and Assessment issued the "Announcement on the Update of the List of Bacteria That Can Be Used in Food and the List of Bacteria That Can Be Used in Infant Food", which adjusted the classification and naming of some bacteria that can be used in food, and renamed "Lactobacillus paracasei" to "Lactobacillus paracasei", which are essentially the same bacteria. Bifidobacterium bifidum i771, the strain was deposited in the General Microbiological Center of China Microbiological Culture Collection Administration on May 20, 2020, with the deposit number CGMCC NO.19855 and the Latin name Bifidobacterium bifidum , first disclosed in a Chinese invention patent with application number “202011361327.3” and titled “Bifidobacterium bifidum i771, separation and purification method and application thereof”; Bifidobacterium longum subspecies longum i772, the strain was deposited on May 20, 2020 at the General Microbiological Center of China Culture Collection Administration, located at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCCNO.19856, and the Latin name is Bifidobacterium longum subsp.longum , which was first disclosed in a Chinese invention patent with application number "202011361316.5" and name "Bifidobacterium longum subspecies longum i772, its separation and purification method and application".

[0016] Example 1

[0017] This embodiment provides an application of a composition having a digestive tract protective and repairing effect, the composition is used to prepare a dairy product having a digestive tract protective and repairing effect, and the effect is verified by experiments, specifically comprising the following steps: S1. Preparation of experimental materials 1) Sample preparation Sample 1: Take 2'-FL, LNnT and Lactobacillus paracasei N1115, add a certain amount of whole milk powder, and prepare a solution with physiological saline (wherein the concentration of 2'-FL is 0.9g / L, the concentration of LNnT is 0.3g / L, and the viable bacteria of Lactobacillus paracasei N1115 is 10 8 CFU / L; converted to human body, it is equivalent to 697.67mg / 100g of 2'-FL, 232.56mg / 100g of LNnT, and 4.8mg / 100g of Lactobacillus paracasei N1115 powder. Among them, the viable bacteria of Lactobacillus paracasei N1115 powder is 5×10 11 CFU / g), reserve; Sample 2: Take the composition, add an equal amount of whole milk powder, and prepare a solution with physiological saline (wherein the concentration of 2'-FL is 0.9 g / L, the concentration of LNnT is 0.3 g / L, and the amount of live bacteria of Lactobacillus paracasei N1115 is 10 6CFU / L; converted to human body, it is equivalent to 697.67mg / 100g of 2'-FL, 232.56mg / 100g of LNnT, and 0.048mg / 100g of Lactobacillus paracasei N1115 powder. Among them, the viable bacteria of Lactobacillus paracasei N1115 powder is 5×10 11 CFU / g), reserve; Sample 3: Take 2'-FL, LNnT and Bifidobacterium bifidum i771, add equal amounts of whole milk powder, and prepare a solution with saline (wherein the concentration of 2'-FL is 0.9 g / L, the concentration of LNnT is 0.3 g / L, and the viable bacterial count of Bifidobacterium bifidum i771 is 10 8 CFU / L; converted to human body, it is equivalent to 697.67mg / 100g of 2'-FL, 232.56mg / 100g of LNnT, and 4.8mg / 100g of Bifidobacterium bifidum i771 powder in milk powder. Among them, the amount of live bacteria in Bifidobacterium bifidum i771 powder is 5×10 11 CFU / g), reserve; Sample 4: Take 2'-FL, LNnT and Bifidobacterium bifidum i771, add equal amounts of whole milk powder, and prepare a solution with physiological saline (wherein the concentration of 2'-FL is 0.9 g / L, the concentration of LNnT is 0.3 g / L, and the viable bacterial count of Bifidobacterium bifidum i771 is 10 6 CFU / L; converted to human body, it is equivalent to 697.67mg / 100g of 2'-FL, 232.56mg / 100g of LNnT, and 0.048mg / 100g of Bifidobacterium bifidum i771 powder. Among them, the amount of live bacteria in Bifidobacterium bifidum i771 powder is 5×10 11 CFU / g), reserve; Sample 5: Take 2'-FL, LNnT and Bifidobacterium longum subsp. longum i772, add equal amounts of whole milk powder, and prepare a solution with saline (wherein the concentration of 2'-FL is 0.9 g / L, the concentration of LNnT is 0.3 g / L, and the viable bacterial count of Bifidobacterium longum subsp. longum i772 is 10 8 CFU / L; converted to human body, it is equivalent to 697.67mg / 100g of 2'-FL, 232.56mg / 100g of LNnT, and 4.8mg / 100g of Bifidobacterium longum subsp. i772 powder. Among them, the amount of viable bacteria in Bifidobacterium longum subsp. i772 powder is 5×10 11 CFU / g), reserve; Sample 6: Take 2'-FL, LNnT and Bifidobacterium longum subsp. longum i772, add equal amounts of whole milk powder, and prepare a solution with saline (wherein the concentration of 2'-FL is 0.9 g / L, the concentration of LNnT is 0.3 g / L, and the viable bacterial count of Bifidobacterium longum subsp. longum i772 is 10 6 CFU / L; converted to human body, it is equivalent to 697.67mg / 100g of 2'-FL, 232.56mg / 100g of LNnT, and 0.048mg / 100g of Bifidobacterium longum subsp. i772 powder. Among them, the amount of viable bacteria in Bifidobacterium longum subsp. i772 powder is 5×10 11 CFU / g), reserve; Sample 7: Take 2'-FL and LNnT, add an equal amount of whole milk powder, and prepare a solution with physiological saline (wherein the concentration of 2'-FL is 0.9 g / L and the concentration of LNnT is 0.3 g / L; converted to the human body, this is equivalent to 697.67 mg / 100 g of 2'-FL and 232.56 mg / 100 g of LNnT in milk powder), and set aside; Sample 8: Take 2'-FL, LNnT and Lactobacillus paracasei JMCC0012, add a certain amount of whole milk powder, and prepare a solution with physiological saline (wherein the concentration of 2'-FL is 0.9g / L, the concentration of LNnT is 0.3g / L, and the viable bacteria of Lactobacillus paracasei JMCC0012 is 10 8 CFU / L; converted to human body, it is equivalent to 697.67mg / 100g of 2'-FL, 232.56mg / 100g of LNnT, and 4.8mg / 100g of Lactobacillus paracasei N1115 powder. Among them, the viable bacteria of Lactobacillus paracasei N1115 powder is 5×10 11 CFU / g), reserve; Sample 9: Take 2'-FL, LNnT and Lactobacillus paracasei JMCC0012, add equal amounts of whole milk powder, and prepare a solution with saline (wherein the concentration of 2'-FL is 0.9 g / L, the concentration of LNnT is 0.3 g / L, and the viable bacterial count of Lactobacillus paracasei JMCC0012 is 10 6 CFU / L; converted to human body, it is equivalent to 2'-FL in milk powder is 697.67mg / 100g, LNnT is 232.56mg / 100g, and Lactobacillus paracasei JMCC0012 powder is 0.048mg / 100g, among which, the viable bacteria of Lactobacillus paracasei JMCC0012 powder is 5×10 11 CFU / g), and set aside.

[0018] 2) Experimental animals C57 suckling mice.

[0019] S2. Experimental methods and results 1) Animal grouping and feeding plan 66 suckling mice were numbered and divided into 11 groups: 9 experimental groups (respectively marked as Group 1 to Group 9), 1 blank group (marked as Blank) and 1 model group (marked as Contorl), with 6 mice in each group. Mixed cages were used (each cage contained mice from different groups) to ensure that the birth and subsequent feeding environment were consistent.

[0020] From birth to day 14 (within 2 weeks), all mice in the 11 groups were fed with breast milk; From the 15th day to the 21st day (week 3), in addition to continuing to feed breast milk, Group 1 to Group 9 were fed with equal amounts of samples 1 to 9 once in the morning and evening, while the blank group and the model group continued to feed breast milk and were fed with equal amounts of normal saline once in the morning and evening; From the 22nd day to the 49th day (the 4th week to the 7th week), Group 1 to Group 9 were fed with feed, and the same amount of samples 1 to 9 were added once in the morning and evening, respectively. The blank group and the model group were fed with feed, and the same amount of normal saline was added once in the morning and evening, respectively. From the 29th day, the feeding method of samples / normal saline was changed from feeding to gavage. After day 50, the feeding regimen was no longer changed (feed + oral feeding sample / saline solution); From the 50th day to the 54th day, for a period of 5 days, 2.5% DSS solution was added to the drinking water of mice in all groups except the blank group.

[0021] 2) Mouse weight test and results The mice were weighed once 2 days before DSS solution treatment and the initial weight was recorded. The mice were weighed every day after DSS solution treatment and the ratio of the weight to the initial weight was recorded. The measurement time period was consistent and the results were as follows: Figure 1 As shown (the 1st to 5th day in the figure is the time of DSS solution treatment, and DSS solution is no longer used for treatment afterwards); Depend on Figure 1 It can be seen that from the 1st to the 5th day, the weight of mice in each group remained basically stable. From the 6th day, under the action of DSS, the weight of mice in most groups began to decrease significantly. The weight loss of Group 1, Group 2 and Group 7 was less, while the weight loss of mice in other groups was greater, and reached the lowest point between the 8th and 9th days. After the 10th day, the weight of mice in each group gradually recovered, especially the mice in Group 1, Group 2 and Group 7, which recovered faster and approached or even exceeded the initial weight after the 11th day. In contrast, the weight of mice in other groups recovered more slowly, and even failed to fully recover to the initial weight. In addition, the weight recovery of the Control group was also relatively slow, indicating the success of the modeling. In summary, the samples of Group 1, Group 2 and Group 7 have a strong protective effect on the weight of mice, indicating that 2'-FL and LNnT, two kinds of human milk oligosaccharides, may have a good anti-DSS effect, which helps to slow down weight loss and speed up recovery; the combination of Lactobacillus paracasei N1115 and these two human milk oligosaccharides has a synergistic effect in resisting DSS, while other strains (such as Bifidobacterium bifidum i771 and Bifidobacterium longum subsp. longum i772, and even other subspecies of Lactobacillus paracasei) do not have a synergistic effect with these two human milk oligosaccharides, and may even hinder the efficacy of human milk oligosaccharides; Group 2 has better effects than Group 1 or Group 7, indicating that Lactobacillus paracasei N1115, 2'-FL and LNnT have a synergistic effect in resisting DSS, and low concentrations of Lactobacillus paracasei N1115 have better effects.

[0022] 3) Mouse DAI scoring and results The disease activity index (DAI) is a comprehensive score based on the percentage of weight loss, stool viscosity, and stool bleeding of the diseased mice. The DAI score was recorded every day after the start of DSS solution treatment. The results are as follows: Figure 2 As shown (the 1st to 5th day in the figure is the time of DSS solution treatment, and DSS solution is no longer used for treatment afterwards); Depend on Figure 2 It can be seen that after DSS treatment, the DAI scores of mice in Group 2 and Group 7 increased slightly, but the peak values ​​were lower (Group 2 was about 2 points, and Group 7 was about 4 points), indicating that the inflammatory response was mild, especially in Group 2, where the DAI score remained at a low level and quickly recovered to 0 points after the end of DSS treatment, showing a significant anti-inflammatory effect; After DSS treatment, the DAI scores of mice in other groups rose rapidly, reaching a peak between the 6th and 8th days (ranging from 7 to 10 points), and then gradually declined, but the recovery speed was slow. At the end of the experiment, the DAI scores of mice in Group 8 and Group 9 had not completely returned to zero, indicating that the inflammatory response was strong and the anti-inflammatory effect was poor. After DSS treatment, the DAI score of the Control group increased significantly and reached the highest value (about 9 points) on the 8th day, and then slowly decreased, but still maintained a high DAI score at the end of the experiment, indicating that the Control group was most sensitive to the DSS-induced inflammatory response. This group was successfully modeled and had a strong inflammatory response.

[0023] 4) Mouse colonoscopy test and results On the 56th day of feeding, 3 mice were selected from each of the 11 groups for colonoscopy and pathological sections after sacrifice. On the 63rd day of feeding, all mice were sacrificed, colonoscopy was performed, and pathological sections were preserved. Group 2 with the smallest inflammatory score and weight change, Group 3 with the largest inflammatory score and weight change, blank group and model group were selected for intestinal mucosal observation. The results are shown in the figure. Figure 3 As shown; Depend on Figure 3 It can be seen that after DSS treatment, the intestinal lining of mice in Group 2 showed slight redness and swelling, and the mucosal damage was relatively mild. At the end of the experiment, the intestinal lining was basically restored, showing a good protective effect on the digestive tract, which indicates that the samples of Group 2 may have a significant effect on resisting DSS-induced damage; After DSS treatment, the intestinal lining of mice in Group 3 was significantly damaged, with ulcers and redness in some areas. Although there was a certain degree of recovery at the end of the experiment, mucosal damage still existed, indicating that the samples in this group had a relatively weak protective effect on the digestive tract. After DSS treatment, the intestinal lining of the mice in the Control group was severely damaged, showing significant redness, swelling and ulcers, and the recovery effect was poor at the end of the experiment, indicating that the tolerance of the Control group to DSS was low. This group was successfully modeled, reflecting the damage effect induced by DSS.

[0024] Example 2 This embodiment provides a composition with digestive tract protection and repair effects, which comprises 0.1 g of Lactobacillus paracasei N1115 powder, 1 kg of 2'-FL and 300 g of LNnT (i.e., a weight ratio of 1:10000:3000), wherein the amount of live bacteria in the Lactobacillus paracasei N1115 powder is 8×10 10 CFU / g.

[0025] Example 3 This embodiment provides a composition with digestive tract protection and repair effects, which comprises 0.02 g of Lactobacillus paracasei N1115 powder, 300 g of 2'-FL and 100 g of LNnT (i.e., a weight ratio of 1:15000:5000), wherein the amount of live bacteria in the Lactobacillus paracasei N1115 powder is 5×109 CFU / g.

[0026] Example 4 This embodiment provides a composition with digestive tract protection and repair effects, which comprises 0.1 g of Lactobacillus paracasei N1115 powder, 1.3 kg of 2'-FL and 400 g of LNnT (i.e., a weight ratio of 1:13000:4000), wherein the amount of viable bacteria of Lactobacillus paracasei N1115 powder is 9×10 11 CFU / g.

[0027] Example 5 This embodiment provides a composition with digestive tract protection and repair effects, which comprises 0.01 g of Lactobacillus paracasei N1115 powder, 140 g of 2'-FL and 35 g of LNnT (i.e., a weight ratio of 1:14000:3500), wherein the amount of live bacteria of Lactobacillus paracasei N1115 powder is 7×10 10 CFU / g.

[0028] Example 6 This embodiment provides a dairy product with digestive tract protection and repair effects, which is prepared by adding the composition described in Example 2 to whole milk powder, with 6.5 kg of the composition added per 1 t of the dairy product.

[0029] Example 7 This embodiment provides a dairy product with digestive tract protection and repair effects, which is prepared by adding the composition described in Example 3 to whole milk powder, with 10 kg of the composition added to every 1 t of the dairy product.

[0030] Example 8 This embodiment provides a dairy product with digestive tract protection and repair effects, which is prepared by adding the composition described in Example 4 to whole milk powder, with 15 kg of the composition added per 1 t of the dairy product.

[0031] Example 9 This embodiment provides a dairy product with digestive tract protection and repair effects, which is prepared by adding the composition described in Example 5 to whole milk powder, with 13 kg of the composition added per 1 t of the dairy product.

Claims

1. A composition having digestive tract protection and repairing effects, characterized in that: The raw materials for making the lactobacillus paracasei N1115, 2'-fucosyllactose and lactose-N-neotetraose in a weight ratio of 1:10000-15000:3000-5000.

2. A composition having digestive tract protection and repairing effects according to claim 1, characterized in that: The viable bacterial count of the Lactobacillus paracasei N1115 is 5×10 9 ~9×10 11 CFU / g.

3. The use of the composition according to claim 1 or 2, characterized in that: The composition is used for preparing probiotic products or dairy products with digestive tract protection and repair effects.

4. A dairy product with digestive tract protection and repair effects, characterized in that: The dairy product is prepared by adding the composition according to claim 1 or 2 to whole milk powder.

5. The dairy product with digestive tract protection and repairing effects according to claim 4, characterized in that: The addition amount of the composition is 6.5-15 kg / t.

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