CN113349375A - Preparation method and application of composition containing animal bifidobacterium F1-7 and krill oil for improving atherosclerotic lipid metabolism - Google Patents
Preparation method and application of composition containing animal bifidobacterium F1-7 and krill oil for improving atherosclerotic lipid metabolism Download PDFInfo
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- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
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- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
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Abstract
The invention belongs to the technical field of food microorganisms and biological medicines, and discloses a preparation method and application effects of a composition for reducing fat and improving atherosclerosis, wherein the composition of bifidobacterium animalis F1-7 and krill oil consists of bifidobacterium animalis F1-7 and krill oil, and the content of the bifidobacterium animalis F1-7 and the krill oil is 108Mixing the CFU/mL animal bifidobacterium F1-7 with 6 mg krill mixed oil in a ratio of 1: 5-1: 10 to prepare the composition. The probiotic krill oil composition provided by the invention can inhibit the accumulation of atherosclerotic lipid and has a very wide application prospect. The inventionAlso relates to a method for preparing the composition of the animal bifidobacterium F1-7 and the krill oil into dripping pills, tablets, granules and capsules for medicines, health-care foods, special medical formula foods and special dietary foods.
Description
Technical Field
The invention belongs to the technical field of food microorganisms, and particularly relates to a preparation method and application effects of solid oral preparations such as capsules, granules, tablets and the like of a composition of lipid-lowering, anti-inflammatory and atherosclerosis-improving bifidobacterium animalis F1-7 and krill oil.
Background
Cardiovascular diseases (CVDs) are one of the leading causes of death in the population worldwide and are closely related to atherosclerosis. Atherosclerosis is a pathologically altered lipid accumulation characterized by the accumulation of blood lipids, particularly cholesterol, in large and medium sized intima of arteries and is a key factor in the development of atherosclerosis. The treatment of the related diseases such as atherosclerosis and the like in the past mostly takes lipid reduction as a main target, and the treatment efficacy is also uneven. According to recent research reports, the imbalance of the intestinal flora has important influence on the development and the progress of atherosclerosis, the intestinal flora is involved in metabolic pathways related to the development of cardiovascular diseases, probiotics and metabolites thereof can improve the formation of atherosclerotic plaques by adjusting the intestinal flora, and probiotics with BSH activity can be involved in bile acid cholesterol metabolic pathways by regulating secondary bile acid products and acting on bile acid metabolism key targets FXR, CYP7A1 and the like, so as to improve the lipid level of organisms. In vitro, in vivo and clinical experiments for alleviating atherosclerosis-related cardiovascular diseases by probiotics are actively carried out, and the analysis of the effects of probiotics on AS and cardiovascular diseases by combining metabonomics becomes a hot spot of current scientific research.
Krill oil is extracted from Antarctic krill (Euphausia superba) and has many health promoting properties such as anti-inflammatory, hypoglycemic and hypolipidemic effects, wherein docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) are abundant, which are important components of omega-3 polyunsaturated fatty acids and have various biological effects including hypolipidemic effects. In recent years, the effect of interaction between active substances and probiotics on disease improvement is in the spotlight, but the specific mechanism of the targeted regulation and control of probiotics involved in disease improvement is not well researched, and the interaction mechanism between probiotics and active substances is not clear. Therefore, screening and developing excellent probiotic and active substance combination products for improving atherosclerosis have important research significance and application value.
In the literature and patents or patent applications that have been published so far, for example, CN109769943A discloses a yogurt food containing sea buckthorn and probiotics that can be used for improving atherosclerosis; CN104172191A discloses a full nutritional formula food which can be used for improving atherosclerosis; CN105852100A discloses a medical formula food for atherosclerosis. In view of the above-mentioned prior art, the common points of these patents or patent applications are that the use of probiotics in combination with various ingredients for preventing or alleviating atherosclerosis, the interaction of probiotics with single active substances for improving diseases is less relevant to research, and the product is made of phoenix-hair-like unicorn. There is currently no report on the combination of probiotics with krill oil as an atherosclerosis-relieving substance. Meanwhile, the reasonable proportion and combination effect and mechanism of the effective probiotics and the krill oil are not effectively verified and explored. Lack of efficacy verification and mechanism disclosure for improving the composition of bifidobacterium animalis F1-7 and krill oil for atherosclerotic animals. The significance of solving the problems and the defects lies in developing the bifidobacterium animalis F1-7 and krill oil composition capable of effectively relieving atherosclerosis, disclosing the mechanism of the bifidobacterium animalis F1-7 and krill oil composition for synergistically relieving atherosclerosis, and providing a reliable theoretical basis and an effective intervention mode for relieving clinical atherosclerosis-related cardiovascular diseases.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a composition for improving bifidobacterium animalis F1-7 and krill oil, a preparation method and application thereof.
The invention is realized by the fact that the composition for improving the bifidobacterium animalis F1-7 and the krill oil consists of the bifidobacterium animalis F1-7F1-7And krill oil (10)8 Movement of CFU/mLBifidobacterium longum F1-7 and 6 mg krill oil in a ratio of 1: 5-1: 10).
The invention aims to provide an application of an animal bifidobacterium F1-7 and krill oil composition for improving atherosclerosis in reducing blood fat and improving atherosclerosis.
The invention has the advantages and positive effects that: the bifidobacterium animalis F1-7 and the bifidobacterium animalis F1-7 in the krill oil composition for improving atherosclerosis have good capacity of regulating lipid metabolism. Apoe against the model of atherosclerosis when Bifidobacterium animalis F1-7 is combined with krill oil-/-Mouse with obvious improvement effect.
The invention only needs a single strain and a single active substance to have good effect of improving atherosclerosis, so the invention can be used as an excellent microbial preparation matched with the traditional medicine for regulating blood fat metabolism and improving lipid plaque accumulation. Compared with the prior art, the method combines the early in vitro and in vivo lipid-lowering effect to quickly and accurately obtain the potential functional strain for relieving atherosclerosis. Therefore, the invention has the following advantages: (1) the strain has outstanding lipid-lowering performance. The bifidobacterium animalis F1-7 provided by the invention has good lipid metabolism regulation capacity and can obviously reduce the aortic plaque accumulation. (2) The composition of bifidobacterium animalis F1-7 and krill oil has a remarkable effect of relieving atherosclerotic plaques. Bifidobacterium animalis F1-7 when combined with krill oil was targeted to Apoe-/-The mouse has obvious improvement effect which is superior to the effect of the two independent effects.
In conclusion, the bifidobacterium animalis F1-7 and krill oil composition provided by the invention has an excellent effect of relieving atherosclerosis, and has a very wide application prospect.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments of the present invention will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
Fig. 1 is a flow chart of a preparation method of the composition for improving bifidobacterium animalis F1-7 and krill oil, which is provided by the embodiment of the invention.
FIG. 2 shows the composition of Bifidobacterium animalis F1-7 and krill oil versus Apoe according to the present invention-/-Schematic representation of oil red microscopic observation of mouse aortic plaque accumulation.
FIG. 3 shows the composition of Bifidobacterium animalis F1-7 and krill oil versus Apoe according to the present invention-/-Schematic view of HE staining observation of mouse liver tissue.
FIG. 4(A) shows the composition of Bifidobacterium animalis F1-7 and krill oil versus Apoe according to the present invention-/-Schematic representation of mouse serum TC levels.
FIG. 4(B) shows the composition of Bifidobacterium animalis F1-7 and krill oil versus Apoe according to the present invention-/-Schematic representation of mouse serum TG levels.
FIG. 4(C) shows the composition of Bifidobacterium animalis F1-7 and krill oil versus Apoe according to the present invention-/-Schematic representation of mouse serum HDL levels.
FIG. 4(D) shows the composition of Bifidobacterium animalis F1-7 and krill oil versus Apoe according to the present invention-/-Schematic of mouse serum LDL levels.
FIG. 5 shows the composition of Bifidobacterium animalis F1-7 and krill oil versus Apoe according to the present invention-/-The content of total bile acid in mouse feces is shown schematically.
FIG. 6 shows the composition of Bifidobacterium animalis F1-7 and krill oil versus Apoe according to the present invention-/-Schematic representation of expression levels of key genes of lipid metabolism in mice.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In order to solve the problems in the prior art, the invention provides a composition for improving bifidobacterium animalis F1-7 and krill oil, a preparation method and application thereof, and the invention is described in detail with reference to the accompanying drawings.
The composition for improving the atherosclerosis bifidobacterium animalis F1-7 and the krill oil provided by the embodiment of the invention consists of bifidobacterium animalis F1-7 and krill oil. The bifidobacterium animalis F1-7 is preserved in China center for type culture Collection, the address is Wuhan university, Wuhan, China, the preservation date is 2020, 12 and 02 months, the preservation number is CCTCC NO: M2020833, and the bifidobacterium animalis subspecies F1-7 are classified and namedBifidobacterium animalis subsp F1-7. The sequence of the 16sRNA of the bifidobacterium animalis F1-7 is as follows: GGGTGGGGGGCGTTCTTACACATGCAGTCGAACGGGATCCCTGGCAGCTTGCTGTCGGGGTGAGAGTGGCGAACGGGTGAGTAATGCGTGACCAACCTGCCCTGTGCACCGGAATAGCTCCTGGAAACGGGTGGTAATACCGGATGCTCCGCTCCATCGCATGGTGGGGTGGGAAATGCTTTTGCGGCATGGGATGGGGTCGCGTCCTATCAGCTTGTTGGCGGGGTGATGGCCCACCAAGGCGTTGACGGGTAGCCGGCCTGAGAGGGTGACCGGCCACATTGGGACTGAGATACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGACGCCGCGTGCGGGATGGAGGCCTTCGGGTTGTAAACCGCTTTTGTTCAAGGGCAAGGCACGGTTTCGGCCGTGTTGAGTGGATTGTTCGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTATCCGGATTTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCCTAACGGTGGATCTGCGCCGGGTACGGGCGGGCTGGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTCACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGGATGCTGGATGTGGGGCCCTTTCCACGGGTCCCGTGTCGGAGCCAACGCGTTAAGCATCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGAAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATGTGCCGGATCGCCGTGGAGACACGGTTTCCCTTCGGGGCCGGTTCACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCGCATGTTGCCAGCGGGTGATGCCGGGAACTCATGTGGGACCGCCGGGGTCAACTCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCATGCTACAATGGCCGGTACAACGCGGTGCGACACGGTGACGTGGGGCGGATCGCTGAAAACCGGTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGGCGGAGTCGCTAGTAATCGCGGATCAGCAACGCCGCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGTAGCACCCGAAGCCGGTGGCCCGACCCTTGTGGGGGGAGCCGTCTAAGGGTAAGAACTCG
As shown in fig. 1, the preparation method of the composition for improving bifidobacterium animalis F1-7 and krill oil provided by the embodiment of the invention comprises the following steps:
s101, culturing animal bifidobacterium F1-7: inoculating animal bifidobacterium F1-7 into a strain-increasing MRS culture medium of a fermentation tank for culture to obtain fermentation liquor;
s102, drying bifidobacterium animalis F1-7: sequentially carrying out pre-cooling treatment, pre-freezing treatment, main drying treatment and analysis drying treatment on bifidobacterium animalis F1-7 before freezing to finish the preparation of a bifidobacterium freeze-dried product;
s103, preparation and packaging of the bifidobacterium animalis F1-7 and krill oil composition: the bifidobacterium animalis F1-7 is mixed with krill oil to prepare the composition of the bifidobacterium animalis F1-7 and the krill oil.
S104, adding auxiliary materials into the composition of the bifidobacterium animalis F1-7 and the krill oil, and preparing into pills, tablets, granules and capsules.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
Example 1 Bifidobacterium animalis F1-7 and krill oil composition have good effect in improving atherosclerosis
The experimental results are shown in fig. 2, the aorta wall of the atherosclerosis model mouse is thickened, hardened and easy to break, calcified plaques are in the blood vessel, the plaques on the inner wall of the aorta are obviously reduced in the bifidobacterium animalis F1-7 group, the krill oil group and the bifidobacterium animalis F1-7 and krill oil composition group, and the aorta wall is firm and elastic. Software analysis of plaque coverage area proves that three intervention groups can effectively improve atherosclerosis caused by high fat diet.
Analysis of HE staining results in liver cases of mice revealed that hepatocytes of AS group mice were swollen, loose cytoplasm and vacuolated with varying size (fig. 3). The Bifidobacterium animalis F1-7, krill oil, Bifidobacterium animalis F1-7 and krill oil composition can improve liver lipid accumulation and relieve liver steatosis. The composition of bifidobacterium animalis F1-7 and krill oil has clear liver cords, obvious liver cell boundary and less intracellular lipid drop deposition. The effect is obviously better than that of animal bifidobacterium F1-7 and krill oil. The prognosis of dried animal bifidobacteria F1-7, although improving liver, was less effective than the krill oil group.
A disorder of serum lipid metabolism in an atherosclerotic model mouse (figure 4) in which the cholesterol level is elevated to 11.82 ± 0.95 mmol/L. After the bacterial strain and the krill oil are respectively dried, the TC content in the serum can be reduced by 30 percent, and when the bacterial strain and the krill oil are mixed and subjected to gastric lavage and drying, the TC content in the serum is reduced by 40 percent. Compared with the model group, the probiotic intervention group and the bifidobacterium animalis F1-7 and krill oil composition group can effectively improve the serum TG content of the atherosclerosis mice, but the krill oil group is not different from the AS group. The three intervention groups can obviously increase the content of HDL in serum of an atherosclerotic mouse and reduce the level of LDL in the serum.
After the probiotics, the krill oil and the mixed dry prognosis are given, the total bile acid content in the excrement of the atherosclerotic mice is obviously increased, and the probiotics intervention and the active substance krill oil are proved to be capable of effectively promoting bile acid metabolism and increasing the discharge of bile acid of the mice so as to participate in the regulation of the lipid level of the mice (figure 5).
Example 2 Bifidobacterium animalis F1-7 and krill oil lipid lowering mechanism of action
The determination of the expression content of the key genes of the lipid metabolism of the atherosclerotic mice is shown in fig. 6, the probiotic intervention can effectively reduce the expression of FXR (p < 0.05) in the intestinal tracts of the model mice, the krill oil intervention effect is better than that of the bifidobacterium animalis F1-7 intervention group, and the mixed intervention of probiotics and active substances can reduce the expression of FXR in the intestinal tracts, but has no significant difference with other two groups. And (3) dry prognosis is given to bifidobacterium F1-7, krill oil and a mixture of the two in the atherosclerotic mouse, and compared with a model group, the three intervention groups can down-regulate the expression content of FGF15 in the intestinal tract of the mouse, and no difference exists among the groups. Measuring the expression content of CYP7A1 which is a key protein for bile acid metabolism in the liver. Compared with a model group, the bifidobacterium animalis F1-7 and the krill oil can effectively improve the expression of CYP7A1, wherein the krill oil group is obviously superior to the bifidobacterium animalis F1-7 group (p is less than 0.05), and the mixed two can be used for gastric lavage of a mouse to up-regulate the expression of the genes, so that the action effect is not different from that of the krill oil group.
Example 3 products and dosage forms
(1) Preparation of bifidobacterium animalis F1-7 and krill oil composition soft capsule
The soft capsule for improving atherosclerosis is prepared by adding one or more of vegetable oil such as soybean oil, corn oil, safflower oil, and olive oil into composition of animal Bifidobacterium F1-7 and krill oil, and using gelatin, glycerol and water as capsule wall material.
(2) Preparation method of bifidobacterium animalis F1-7 and krill oil composition dropping pill
Adding one or more of stearic acid, glyceryl monostearate, beeswax and hydrogenated vegetable oil as matrix into the composition of animal Bifidobacterium F1-7 and krill oil, and using water or ethanol with different concentrations as condensing medium to prepare the dripping pill for improving cognitive dysfunction.
(3) Preparation method of solid oral preparation such as Bifidobacterium animalis F1-7 and krill oil composition capsule, granule, and tablet
The composition of animal bifidobacterium F1-7 and krill oil is added with one or more of a-cyclodextrin, beta-dextrin, sodium carboxymethylcellulose, gelatin, povidone, PEG-4000, PEG-6000, etc., and can be prepared into solid oral preparations such as capsules, granules, tablets, etc. for improving memory and cognitive dysfunction by one or more of inclusion, microencapsulation and solid dispersion technologies.
The invention relates to a single strain plus a single active substance only, which has good atherosclerosis relieving effect. By atherosclerosis model Apoe-/-Mice verify that the composition of bifidobacterium animalis F1-7 and krill oil with the lipid-lowering function can effectively relieve the accumulation of atherosclerotic plaques. Therefore, the invention has the following advantages: 1. the Bifidobacterium animalis F1-7 combined with krill oil has effects of improving atherosclerosis, reducing blood lipid, and treating atherosclerosis model Apoe-/-Mouse with obvious improvement effect. 2. The interaction of bifidobacterium animalis F1-7 and krill oil can promote bile acid metabolism to regulate the lipid level of the organism through FXR/FGF15/CYP7A1 pathway and reduce the formation of plaquesAnd is superior to the single action of the two.
Sequence listing
<110> China oceanic university
<120> preparation method and application of composition containing animal bifidobacterium F1-7 and krill oil for improving atherosclerotic lipid metabolism
<130> 2021
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1444
<212> DNA
<213> Bifidobacterium animalis subsp. lactis strain
<400> 1
gggtgggggg cgttcttaca catgcagtcg aacgggatcc ctggcagctt gctgtcgggg 60
tgagagtggc gaacgggtga gtaatgcgtg accaacctgc cctgtgcacc ggaatagctc 120
ctggaaacgg gtggtaatac cggatgctcc gctccatcgc atggtggggt gggaaatgct 180
tttgcggcat gggatggggt cgcgtcctat cagcttgttg gcggggtgat ggcccaccaa 240
ggcgttgacg ggtagccggc ctgagagggt gaccggccac attgggactg agatacggcc 300
cagactccta cgggaggcag cagtggggaa tattgcacaa tgggcgcaag cctgatgcag 360
cgacgccgcg tgcgggatgg aggccttcgg gttgtaaacc gcttttgttc aagggcaagg 420
cacggtttcg gccgtgttga gtggattgtt cgaataagca ccggctaact acgtgccagc 480
agccgcggta atacgtaggg tgcgagcgtt atccggattt attgggcgta aagggctcgt 540
aggcggttcg tcgcgtccgg tgtgaaagtc catcgcctaa cggtggatct gcgccgggta 600
cgggcgggct ggagtgcggt aggggagact ggaattcccg gtgtaacggt ggaatgtgta 660
gatatcggga agaacaccaa tggcgaaggc aggtctctgg gccgtcactg acgctgagga 720
gcgaaagcgt ggggagcgaa caggattaga taccctggta gtccacgccg taaacggtgg 780
atgctggatg tggggccctt tccacgggtc ccgtgtcgga gccaacgcgt taagcatccc 840
gcctggggag tacggccgca aggctaaaac tcaaagaaat tgacgggggc ccgcacaagc 900
ggcggagcat gcggattaat tcgatgcaac gcgaagaacc ttacctgggc ttgacatgtg 960
ccggatcgcc gtggagacac ggtttccctt cggggccggt tcacaggtgg tgcatggtcg 1020
tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa ccctcgccgc 1080
atgttgccag cgggtgatgc cgggaactca tgtgggaccg ccggggtcaa ctcggaggaa 1140
ggtggggatg acgtcagatc atcatgcccc ttacgtccag ggcttcacgc atgctacaat 1200
ggccggtaca acgcggtgcg acacggtgac gtggggcgga tcgctgaaaa ccggtctcag 1260
ttcggatcgc agtctgcaac tcgactgcgt gaaggcggag tcgctagtaa tcgcggatca 1320
gcaacgccgc ggtgaatgcg ttcccgggcc ttgtacacac cgcccgtcaa gtcatgaaag 1380
tgggtagcac ccgaagccgg tggcccgacc cttgtggggg gagccgtcta agggtaagaa 1440
ctcg 1444
Claims (6)
1. The bifidobacterium animalis F1-7 is characterized in that the bifidobacterium is preserved in China center for type culture collection, the address is Wuhan university in Wuhan, China, the preservation date is 2020, 12 months and 02 days, the preservation number is CCTCC NO: M2020833, and the bifidobacterium animalis subspecies F1-7 are classified and namedBifidobacterium animalis subsp F1-7。
2. The composition for relieving atherosclerosis bifidobacterium animalis F1-7 and krill oil is characterized in that the composition for relieving atherosclerosis bifidobacterium animalis F1-7 and krill oil consists of bifidobacterium animalis F1-7 and krill oil.
3.108Mixing the CFU/mL animal bifidobacterium F1-7 with 6 mg krill mixed oil in a ratio of 1: 5-1: 10 to prepare the composition.
4. The use of the composition for improving atherosclerosis according to claim 2, wherein the composition of bifidobacterium animalis F1-7 and krill oil has a function of significantly improving atherosclerosis after the intervention of the composition in mice for 8 weeks.
5. The bifidobacterium animalis F1-7 and krill oil composition as claimed in claim 2 has the function of remarkably improving atherosclerosis, and the action mechanism of the composition is that the composition acts on FXR/FGF15 pathway to promote bile acid metabolism efflux and improve lipid metabolism, so that the composition has the function of relieving atherosclerosis.
6. Bifidobacterium animalis F1-7 and krill oil composition according to claim 2, wherein: the composition can be added with health food, special medical formula food, and special dietary food acceptable adjuvants, and made into dripping pill, tablet, granule, and capsule for use in medicine, health food, special medical formula food, and special dietary food.
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