Compound composition of bifidobacterium bifidum i771 as well as application and product thereof

Abstract

The invention relates to a compound composition of bifidobacterium bifidum i771 as well as application and a product of the compound composition. The compound composition is prepared from bifidobacterium bifidum i771, choline and phosphatidylserine. The active bifidobacterium bifidum i771 and a composite product containing the active bifidobacterium bifidum i771, choline and phosphatidylserine have a remarkable effect of improving cognition.

Description

Technical Field

[0001] The present invention relates to the field of microbial engineering technology, and more particularly to a compound composition comprising Bifidobacterium bifidum i771, and more particularly to a composition capable of improving cognitive function and its application. Background Art

[0002] Alzheimer's disease is a major disease that troubles the health of the elderly in my country. Due to the characteristics of Alzheimer's disease, which is insidious and progressive, only 15%-20% of patients in my country have been diagnosed and treated, and more than 80% of patients have never been diagnosed and treated. At the same time, drug treatment mainly relieves symptoms and delays the progression of the disease. In the case of large limitations in the effect of drug treatment, non-drug intervention in the early stage of the disease is very necessary.

[0003] The "gut-brain axis" has been confirmed as a research hotspot by many scholars. It is theoretically supported and feasible to screen probiotics or probiotic combinations through intestinal probiotic research to intervene in brain function and achieve the effect of improving cognition and alleviating Alzheimer's disease. Summary of the invention

[0004] The following is a summary of the subject matter described in detail herein. This summary is not intended to limit the scope of the claims.

[0005] The first aspect of the present application provides a composite composition, which contains Bifidobacterium bifidum i771 and can improve cognitive function, and can be used as an auxiliary support supplement for daily health care of the elderly and early intervention of Alzheimer's disease. Among them, Bifidobacterium bifidum i771 has been deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on May 20, 2020, with an address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, with a deposit number of CGMCCNO.19855, and the Latin name is Bifidobacterium bifidum.

[0006] In an exemplary embodiment, the composite composition comprises Bifidobacterium bifidum i771, choline and phosphatidylserine.

[0007] In an exemplary embodiment, the mass ratio of Bifidobacterium bifidum i771, choline and phosphatidylserine is 1:(5-20):(5-20).

[0008] In an exemplary embodiment, the number of viable bacteria per unit dose of Bifidobacterium bifidum i771 is 5×10 10 -5×10 11 CFU.

[0009] In an exemplary embodiment, the daily intake of Bifidobacterium bifidum i771 is 5×107 CFU-5×10 9 CFU.

[0010] In an exemplary embodiment, the daily intake of Bifidobacterium bifidum i771 is 1×10 9 CFU or 5 × 10 9 CFU.

[0011] In an exemplary embodiment, the daily intake of Bifidobacterium bifidum i771, choline and phosphatidylserine is 5×10 7 -5×10 9 CFU, 60-120mg and 60-120mg;

[0012] Preferably, the daily intake of Bifidobacterium bifidum i771, choline and phosphatidylserine is 1×10 9 CFU, 94.5mg and 99.4mg.

[0013] The second aspect of the present application provides a product, which comprises the above-mentioned compound composition.

[0014] In an exemplary embodiment, the product is a food, a health product, a probiotic, a medicine or a pharmaceutical composition.

[0015] In an exemplary embodiment, the product is formula milk powder for middle-aged and elderly people.

[0016] In one exemplary embodiment, the product is an instant probiotic powder.

[0017] The third aspect of the present application provides an application of Bifidobacterium bifidum i771 or the above-mentioned compound composition or the above-mentioned product containing the same, wherein the application includes the application of Bifidobacterium bifidum i771 or the above-mentioned compound composition or the above-mentioned product in the preparation of a product having improved cognitive function.

[0018] In an exemplary embodiment, the amount of the Bifidobacterium bifidum i771 or the compound composition added to the product having the function of improving cognitive function is 0.2% to 13% by weight.

[0019] In an exemplary embodiment, the number of viable bacteria per unit dose of Bifidobacterium bifidum i771 is 5×10 10 -5×10 11 CFU.

[0020] In an exemplary embodiment, the daily intake of Bifidobacterium bifidum i771 is 5×10 7 CFU-5×10 9 CFU.

[0021] In an exemplary embodiment, the daily intake of Bifidobacterium bifidum i771 is 1×10 9 CFU or 5 × 10 9 CFU.

[0022] In an exemplary embodiment, improving cognitive function includes preventing, alleviating and / or improving cognitive function decline in an individual.

[0023] In an exemplary embodiment, the individual is diagnosed with Alzheimer's disease or early related symptoms or childhood intellectual disability.

[0024] Compared with the existing related technologies, this application has the following effects:

[0025] Active Bifidobacterium bifidum i771 and the compound product containing active Bifidobacterium bifidum i771, choline and phosphatidylserine have significant effects on improving cognition. Among them, the compound product containing active Bifidobacterium bifidum i771, choline and phosphatidylserine has more significant effects on improving cognitive impairment.

[0026] Other features and advantages of the present application will be described in the following description, and partly become apparent from the description, or be understood by implementing the present application. Other advantages of the present application can be realized and obtained by the schemes described in the description and the drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The accompanying drawings are used to provide an understanding of the technical solution of the present application and constitute a part of the specification. Together with the embodiments of the present application, they are used to explain the technical solution of the present application and do not constitute a limitation on the technical solution of the present application.

[0028] Figure 1 is the percentage of time each group of mice stayed in the original platform position area in the first water maze test in Example 1;

[0029] Figure 2 is the number of times each group of mice crossed the original platform location area in the first water maze test in Example 1;

[0030] Figure 3 is the percentage of time each group of mice stayed in the original platform position area in the second water maze test in Example 1;

[0031] Figure 4 It is the number of times each group of mice crossed the original platform location area in the second water maze test in Example 1. DETAILED DESCRIPTION

[0032] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] The raw materials used in this application are all conventional products on the market.

[0034] Unless otherwise specified, the materials and reagents used in the examples of the present invention can be obtained from commercial sources. Experimental methods without specific conditions in the examples are usually carried out under conventional conditions or under conditions recommended by the manufacturer.

[0035] In the embodiment:

[0036] Bifidobacterium bifidum i771 was deposited in the General Microbiological Center of China National Committee for the Administration of Microbiological Culture Collection on May 20, 2020, at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCCNO.19855 and the Latin name Bifidobacterium bifidum;

[0037] The commercial strains Bifidobacterium bifidum JM-1114 and Bifidobacterium bifidum JM-1115 were provided by Junlebao Microbiological Collection Center (JMCC) and isolated from the intestines of healthy infants.

[0038] Example 1

[0039] 1) Experimental Materials

[0040] Sample preparation

[0041] Preparation method of Bifidobacterium bifidum compound product: choline (15 mg / kg) + phosphatidylserine (16 mg / kg) + active Bifidobacterium bifidum (2 mg / kg), oral gavage volume is 0.5 mL / head, equivalent to the daily intake of 94.5 mg + 99.4 mg + 1×10 9 CFU

[0042] Experimental animals

[0043] C57BL / 6 mice, male, 14 weeks old.

[0044] APP / PS1 mice, male, 14 weeks old.

[0045] The experimental animals were provided by Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd. (China), certificate: SCXK(Su)2018-0008.

[0046] Instruments, consumables and reagents

[0047] Disposable 5 ml syringe (Shanghai Kangdeli, China), electronic analytical balance (Precisa12A, Precisa, Switzerland), depyrogenation ultrapure water system (Millipo3150, Millipore, USA), mouse gavage needle (Zhongke Life Science, China), electric constant temperature water bath (HW-SY11-K P2, Beijing Changfeng Instrument Co., Ltd., China), centrifuge tube (1.5 ml, Wuxi Nice Life Science Co., Ltd., China), water maze apparatus (ACT-200A, Beijing Youcheng Biotechnology Co., Ltd., China).

[0048] Edaravone (M70800-5G, Sigma-Aldrich, USA).

[0049] 2) Experimental methods

[0050] Animal grouping and oral administration

[0051] 1. The mice were raised in an SPF environment with an ambient temperature of 23±2℃, a humidity of 55±5%, and a cycle of 12 hours of light and 12 hours of darkness. After 7 days of adaptive feeding, 100 mice were divided into 10 groups: blank group, model group, positive drug group, experimental group 1, experimental group 2, experimental group 3, experimental group 4, experimental group 5, experimental group 6, and experimental group 7, with 10 mice in each group. Among them, the blank group used C57BL / 6 male mice without any treatment, and the other groups used APP / PS1 congenital cognitive impairment male mice.

[0052] 2. The mice in each group were gavaged daily. The mice in the blank group and the model group were given an equal volume of normal saline daily. The mice in the positive drug group were given 3 mg / kg of edaravone daily, with the solvent being normal saline, once a day for 42 consecutive days.

[0053] 3. Experimental group 1 was given active Bifidobacterium bifidum JM-1114 by gavage every day at a dose of 10 mg / kg, which is equivalent to a daily intake of 5×10 9 CFU; Experimental group 2 was given active Bifidobacterium bifidum JM-1115 by gavage every day, the dose was 10 mg / kg, which is equivalent to a daily intake of 5×10 9 CFU; Experimental group 3 was given active Bifidobacterium bifidum i771 by gavage every day, the dose was 10 mg / kg, which is equivalent to a daily intake of 5×10 9 CFU. Once a day for 42 consecutive days.

[0054] 4. Experimental group 4 was given a composite product by gavage every day. The specific ingredients were choline + PS (phosphatidylserine) + active Bifidobacterium bifidum JM-1114, with a dose of 15 mg / kg + 16 mg / kg + 2 mg / kg, which is equivalent to a daily intake of 94.5 mg + 99.4 mg + 1×10 9 CFU; Experimental group 5 was given a composite product by gavage every day, with the specific ingredients of choline + PS (phosphatidylserine) + active Bifidobacterium bifidum JM-1115, and the dosage was 15mg / kg + 16mg / kg + 2mg / kg, which is equivalent to a daily intake of 94.5mg + 99.4mg + 1×10 9 CFU. Once a day for 42 consecutive days. Experimental group 6 was given a composite product by gavage every day. The specific ingredients were choline + PS (phosphatidylserine) + active Bifidobacterium bifidum i771, with doses of 15mg / kg + 16mg / kg + 2mg / kg, which is equivalent to a daily intake of 94.5mg + 99.4mg + 1×10 9 CFU.

[0055] 5. Experimental group 7 was given choline and phosphatidylserine only by oral gavage daily, with the dosage of 15mg / kg+16mg / kg respectively, which is equivalent to a daily intake of 94.5mg+99.4mg for the human body.

[0056] Water maze experimental training and testing

[0057] 1. 24 hours after the last oral administration, each group began water maze training: 4 training sessions per day for each mouse, each time starting from a different quadrant of the water maze and entering the swimming pool. Training was terminated when the mouse found the underwater platform or lasted for 60 seconds. Each training session was 15 minutes apart, and training was continued for 4 days.

[0058] 2. After 4 days of training, the water maze test was started on the 5th day: each mouse entered the swimming pool (without platform) 4 times, starting from a different quadrant of the water maze each time. The movement trajectory of the mice within 120 s was recorded, and the proportion of the time each group of mice stayed in the original platform position area within the 120 s test time, as well as the number of times they crossed the original platform position area, were analyzed.

[0059] 3. After completing the first water maze training and testing, each group of mice continued to be gavaged with the tested probiotics and compound products once a day for 2 consecutive weeks.

[0060] 4. After the second gavage for 2 weeks, repeat the water maze training and testing of steps 1 and 2 above for each group, and record and analyze again the proportion of time each group of mice stayed in the original platform position area within the 120s test time in the second water maze test, as well as the number of times they crossed the original platform position area.

[0061] 3) Experimental results

[0062] Evaluation of the efficacy of active probiotics and their compound products in improving cognition

[0063] 1. Active Bifidobacterium bifidum i771 and its combination products with choline and phosphatidylserine have the effect of improving cognitive impairment. Specifically, it can increase the proportion of time that APP / PS1 mice with congenital cognitive impairment stay in the original platform position area in the water maze test (p<0.01), and increase the number of times the original platform position area is crossed (p<0.01). Among them, after the first experiment, the proportion of time that mice in the active Bifidobacterium bifidum i771 group and the i771 combination group (choline + phosphatidylserine + active Bifidobacterium bifidum) stay in the original platform position area was significantly different from that in the model group. See details. Figure 1 to Figure 2 ,in, Figure 1 to Figure 2 Compared with the blank control group, **p<0.01; compared with the model control group, #p<0.05, ##p<0.01.

[0064] 2. To verify the efficacy, continue feeding for 2 weeks and then conduct the same test. The results are shown in Figure 3 to Figure 4 Among them, compared with the blank control group, the active Bifidobacterium bifidum i771 group and the i771 composition group, **p<0.01; compared with the model control group, #p<0.05, ##p<0.01.

[0065] 4) Experimental conclusion

[0066] Under the experimental conditions, active Bifidobacterium bifidum i771 and the compound product containing active Bifidobacterium bifidum i771, choline and phosphatidylserine have significant effects on improving cognition. Among them, the compound product containing active Bifidobacterium bifidum i771, choline and phosphatidylserine has a more significant effect on improving cognitive impairment.

[0067] It has been verified that the composition ratio within the scope of the present application has a similar effect to that of Example 1.

[0068] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limitations on the present application. Ordinary technicians in this field can change, modify, replace and modify the above embodiments within the scope of the present application.

Claims

1. A composite composition, characterized in that: The composite composition comprises Bifidobacterium bifidum i771, choline and phosphatidylserine. The Bifidobacterium bifidum i771 has been deposited in the General Microbiological Center of the China National Microbiological Culture Collection Administration on May 20, 2020, with a deposit number of CGMCCNO.19855, and its Latin name is Bifidobacteriumbifidum.

2. The composite composition according to claim 1, characterized in that The mass ratio of the Bifidobacterium bifidum i771, choline and phosphatidylserine is 1:(5-20):(5-20).

3. The composite composition according to claim 1 or 2, characterized in that: The number of viable bacteria per unit dose of Bifidobacterium bifidum i771 is 5×10 10 -5×10 11 CFU.

4. The composite composition according to claim 1 or 2, characterized in that: The daily intake of choline and phosphatidylserine of Bifidobacterium bifidum i771 is 5×10 7 -5×10 9 CFU, 60-120mg and 60-120mg.

5. The composite composition according to claim 4, characterized in that: The daily intake of Bifidobacterium bifidum i771, choline and phosphatidylserine is 1×10 9 CFU, 94.5mg and 99.4mg.

6. A product, characterized in that The product comprises the compound composition according to any one of claims 1 to 5, and optionally, the product is a food, a health product, a probiotic, a medicine or a pharmaceutical composition.

7. The product according to claim 6, characterized in that The product is formula milk powder for middle-aged and elderly people; or the product is instant probiotic powder.

8. Use of Bifidobacterium bifidum i771 or the composite composition according to any one of claims 1 to 5 or the product according to claim 6 or 7 in the preparation of a product having the function of improving cognitive function.

9. The use according to claim 8, characterized in that: In terms of weight percentage, the amount of the Bifidobacterium bifidum i771 or the compound composition added to the product having the function of improving cognitive function is 0.2% to 13%.

10. The use according to claim 8 or 9, characterized in that: Improving cognitive function includes preventing, alleviating and / or improving cognitive function decline of an individual; Optionally, the individual is diagnosed with Alzheimer's disease or early related symptoms or childhood intellectual disability.

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