CN112690456B - Preparation method and application of animal bifidobacterium F1-7 and krill oil composition for improving atherosclerosis inflammation - Google Patents
Preparation method and application of animal bifidobacterium F1-7 and krill oil composition for improving atherosclerosis inflammation Download PDFInfo
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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, resisting inflammation and improving atherosclerosis, wherein the composition consists of bifidobacterium animalis F1-7 and krill oil; the preparation method of the composition for improving atherosclerosis comprises the preparation of tablets, granules and capsules of the composition of animal bifidobacterium F1-7 and krill oil. The probiotic composition provided by the invention has an excellent effect of improving the inflammatory response of atherosclerosis.
Description
Technical Field
The invention belongs to the technical field of food microorganisms, and particularly relates to a preparation method and application effects of tablets, granules and capsules of a composition of lipid-lowering, anti-inflammatory and atherosclerosis-improving probiotics and krill oil.
Background
Atherosclerosis is closely related to the occurrence and progression of cardiovascular disease, and is accompanied by serious complications. The existing research considers that atherosclerosis is a complex metabolic disorder process, and the occurrence and the development of the atherosclerosis are caused by the combined action of various risk factors on the basis of genetic and environmental factors. The development of atherosclerotic plaques involves abnormal inflammatory cell recruitment, foam cell formation, smooth muscle cell proliferation, extracellular matrix synthesis, reactive oxygen species production, and arterial remodeling, among others. Among these changes, inflammation plays a key role in the development and progression of atherosclerosis. The study of probiotics on improving atherosclerosis by regulating lipid metabolism is more, but the study from the direction of inflammatory pathway is still less, and the relationship between probiotics and atherosclerosis inflammatory reaction cannot be systematically elucidated.
Krill Oil (KO) extracted from antarctic krill (Euphausia superba) is rich in unsaturated fatty acids such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), and plays a key role in regulating lipid metabolism inflammatory response. KO also contains a dose of astaxanthin which is believed to be responsible for the anti-inflammatory, analgesic and hypolipidemic properties of krill oil in human subjects. Krill oil has been found to improve the intestinal flora and metabolic products, for example, krill oil has been found to contribute to its anti-inflammatory activity by inhibiting key metabolites of histidine metabolism in the host and microorganisms. 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.
The common point of the prior patents or patent applications is that the probiotics are mixed with a plurality of components to prevent or relieve atherosclerosis, the interaction of the probiotics and the single active substances has less research on the improvement of diseases, and the related mechanism is not clear. There is currently no report on the atherosclerotic inflammatory response alleviating substance of probiotic bacteria in combination with krill oil.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a preparation method and application of a composition, a tablet, a granule and a capsule for improving atherosclerosis, and particularly relates to a composition of animal bifidobacterium F1-7 and krill oil capable of remarkably relieving an inflammatory reaction of atherosclerosis and application of the composition.
The composition for improving atherosclerosis is prepared by mixing bifidobacterium animalis F1-7 and krill oil (a bacterial solution of bifidobacterium animalis F1-7 and the krill oil in a ratio of 1: 5-1: 10, and the concentrations of the two are 10 8 CFU/mL and 6 mg).
The invention aims to provide application of oral preparations such as tablets, granules and capsules of composition for improving atherosclerosis in resisting inflammatory reaction and improving atherosclerosis, which is characterized in that bifidobacterium animalis F1-7 is combined with krill oil and krill oil to reduce inflammatory and relieve atherosclerosis.
The invention also aims to provide a mechanism of oral preparations such as tablets, granules and capsules for improving atherosclerosis in resisting inflammatory response and improving atherosclerosis, which is characterized in that the oral preparations act on a TLR4/NF kB pathway, the expression of tumor necrosis factor-alpha (TNF-alpha) and interleukin 1 beta (IL-1 beta) is reduced, the inflammatory response is relieved, and the effect of relieving atherosclerosis is further achieved.
The complex formulation of claim 2, wherein: the composition is added with auxiliary materials acceptable for health-care food, special medical formula food and special dietary food, and is prepared into oral preparations such as tablets, granules, capsules and the like for commercial production
The invention only needs a single strain plus a single active substance to have good effect of improving atherosclerosis, thus being capable of being used as an excellent microbial preparation matched with a traditional medicine for regulating inflammatory response and improving lipid plaque accumulation. Therefore, the invention has the following advantages: (1) the strain has outstanding anti-inflammatory performance. The animal bifidobacterium F1-7 provided by the invention has good capability of improving inflammation related to atherosclerosis. (2) Composition for relieving atherosclerosisThe effect of eliminating the speckles is remarkable. When Bifidobacterium animalis F1-7 and krill oil were combined, the combination was directed against Apoe -/- The mouse has obvious improvement effect which is superior to the effect of the two independent effects.
In conclusion, the probiotic composition provided by the invention has an excellent effect of relieving the inflammatory reaction of 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 method for preparing an improved atherosclerotic composition product provided by an embodiment of the present invention.
FIG. 2 shows the combination of Bifidobacterium animalis F1-7 and krill oil for Apoe according to the present invention -/- Schematic representation of aortic cross-section HE of mice.
FIG. 3 shows the combination of Bifidobacterium animalis F1-7 and krill oil for Apoe according to the present invention -/- A graph of the level of the mouse serum inflammatory factor IL-1 β.
FIG. 4 shows the combination of Bifidobacterium animalis F1-7 and krill oil for Apoe according to the present invention -/- A graph of the serum inflammatory factor TNF- α levels in mice.
FIG. 5 shows the combination of Bifidobacterium animalis F1-7 and krill oil for Apoe according to the present invention -/- A schematic diagram of the organization of the expression of the related index of the aortic inflammatory factor of the mouse.
FIG. 6 shows the combination of Bifidobacterium animalis F1-7 and krill oil for Apoe according to the present invention -/- Schematic representation of expression levels of key genes in the inflammatory response 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 atherosclerosis, a preparation method and application thereof, and the invention is described in detail with reference to the accompanying drawings.
The composition for improving atherosclerosis provided by the embodiment of the invention consists of animal bifidobacterium F1-7 and krill oil. The Bifidobacterium animalis F1-7 is preserved in China center for type culture Collection, with the preservation date of 2020, 12 months and 02 days, the preservation number of M2020833, and classification name of Bifidobacterium animalis subspecies F1-7Bifidobacterium animalis subspF1-7. 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 improved atherosclerosis composition provided by the embodiment of the invention comprises the following steps: s101, preparing animal bifidobacterium freeze-dried powder: inoculating animal bifidobacterium into a bacterium-enriched MRS culture medium of a fermentation tank for culture, and sequentially carrying out pre-cooling treatment, pre-freezing treatment, main drying treatment and analysis drying treatment on fermentation liquor before freezing to finish the preparation of a bifidobacterium freeze-dried product; s102, preparing a complex preparation of bifidobacterium animalis F1-7 and krill oil: will 10 8 Mixing CFU/mL animal bifidobacterium F1-7 with 6mg krill mixed oil in a ratio of 1: 5-1: 10 to prepare a composite preparation; s103 bifidobacterium animalis F1-7 and krill oil compound preparation intervention Apoe -/- The mouse measures the inflammatory reaction index and the related metabolic gene expression; s104, adding auxiliary materials into the mixture of the animal bifidobacterium F1-7 and the krill oil to prepare solid oral preparations such as tablets, granules, capsules and the like.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
The technical solution of the present invention is further described with reference to the following examples.
Example 1 Effect of Bifidobacterium animalis F1-7 and Krill oil mixture on improving Atherosclerosis
HE staining of cardiovascular tissue (FIG. 2), the atherosclerotic group Aope -/- The cardiovascular endothelial cells of the mice are widely damaged and shed, lipid deposition occurs, the cytoplasm vacuoles of the smooth muscle cells are formed, the cytoplasm is loose and lightly stained, the arrangement of the smooth muscle cells is disordered, and large plaque-like substances are obviously visible on the inner wall of the blood vessel. A small amount of plaques are observed in krill oil and attached to the inner wall, the cardiovascular intima of mice is slightly damaged, smooth muscle cells are arranged regularly and are proliferated, the cell nucleus density is increased, and some smooth muscle cells are changed in vacuole shape. The probiotic group, the animal bifidobacterium F1-7 and the krill oil combined group have no plaque deposition basically, the mouse heart artery intima is smooth, the endothelial cells are complete, the subintimal space is small, the smooth muscle cells are arranged relatively regularly, and the cytoplasm is dyed in red.
Example 2 anti-inflammatory Effect of Bifidobacterium animalis F1-7 and Krill oil mixture
The level of inflammatory changes in the body of an atherosclerotic mouse is measured, and the animal bifidobacterium F1-7, krill oil and a mixture of the animal bifidobacterium F1-7 and the krill oil can effectively improve the serum inflammatory level of the mouse. The bifidobacterium animalis F1-7, krill oil and bifidobacterium animalis F1-7 and krill oil combination significantly reduced the levels of IL-1 β (fig. 3) and TNF- α (fig. 4) in serum compared to the AS group with no statistical difference between the groups.
TLR4 immunohistochemical staining (figure 5) shows that compared with the atherosclerosis model group AS, the Bifidobacterium animalis F1-7, the krill oil and the TLR4 positive area of the mixture group are obviously reduced, wherein the effect of the combination of the Bifidobacterium animalis F1-7 and the krill oil is better than that of the other two groups. The immunohistochemical result of MYD88 is consistent with that of TLR4, three groups of MYD88 can reduce the expression content of aortic sinus MYD88 and reduce the transduction of proinflammatory signals, and the action effect of animal bifidobacterium F1-7 and krill oil is optimal. The results of the grouping of NF κ B show that three groups all improved inflammatory expression in atherosclerotic mice.
Example 3 anti-inflammatory mechanism of Bifidobacterium animalis F1-7 and krill oil mixture
The expression of genes associated with inflammatory response in the intestine was determined (FIG. 6). Compared with the AS group, the Bifidobacterium animalis F1-7, the krill oil and the mixture thereof can obviously reduce the expression of TNF-alpha. Wherein, the action effect of the bifidobacterium animalis F1-7 and the krill oil is obviously better than that of the other two groups. After the probiotic bacteria, krill oil and mixture were given a dry pretreatment, the expression of IL-1 β was significantly reduced with no difference between groups. The intervention group can also effectively reduce the expression of key genes on a TLR4/MYD 88/NF-kB pathway, thereby inhibiting the levels of downstream inflammatory factors TNF-alpha and IL-1 beta. The effect of bifidobacterium animalis F1-7 and krill oil on regulating TLR4 and MYD88 is not different, and the down-regulation effect of the combination of bifidobacterium animalis F1-7 and krill oil is most obvious. In the down-regulation of NF κ B, the results for bifidobacterium animalis F1-7 and krill oil were not different and better than the combination group (p < 0.05). Summarizing the anti-inflammatory effects of the three intervention groups, the bifidobacterium animalis F1-7, the krill oil and the mixture of the two can effectively improve the effect of the combined group in the inflammatory reaction better than the effect of the two alone. Researches find that the interaction between the animal bifidobacterium F1-7 and krill oil can achieve a better anti-inflammatory effect and improve the inflammatory response related to atherosclerosis.
Example 4 products and dosage forms
(1) Bifidobacterium animalis F1-7 and krill oil mixture tablet
The mixture of animal bifidobacterium F1-7 and krill oil is mixed with raw auxiliary materials in the field of tablets, such as glucose, lactose, microcrystalline cellulose, magnesium stearate, skimmed milk powder, etc., and is prepared into a tablet preparation by a conventional preparation technology.
(2) Bifidobacterium animalis F1-7 and krill oil mixture granule
The mixture of bifidobacterium animalis F1-7 and krill oil is mixed with auxiliary materials such as milk powder, starch, conventional sweetener, essence and the like to produce.
(3) Bifidobacterium animalis F1-7 and krill oil mixture capsule
The mixture of the bifidobacterium animalis F1-7 and the krill oil is prepared into a capsule product by a capsule machine.
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 mixture of the probiotic animal bifidobacterium F1-7 and the krill oil can effectively relieve the accumulation of atherosclerotic plaques. Therefore, the invention has the following advantages: 1. bifidobacterium animalisBifidobacterium animalissubspF1-7 has effect in improving atherosclerosis, and has antiinflammatory effect. 2. The composition has remarkable effect of reducing atherosclerotic plaques. Apoe for atherosclerosis model of porridge soup after combination of Bifidobacterium animalis F1-7 and krill oil -/- Mouse with obvious improvement effect. The interaction of the bifidobacterium animalis F1-7 and the krill oil can effectively improve the inflammatory response of the organism by acting on a TLR4/MYD 88/NF-kB channel, reduce the formation of plaques and is better than the single action of the two.
Claims (1)
1. Improvement of atherosclerosisThe composition for improving atherosclerosis is characterized by comprising bifidobacterium animalis F1-7 and krill oil, wherein the concentration of a bacterial solution of the bifidobacterium animalis F1-7 is 10 8 CFU/mL, the dosage of krill oil is 6mg, and the ratio of bifidobacterium animalis F1-7 to the krill oil is 1: 5-1: 10; the preservation number of the bifidobacterium animalis F1-7 is CCTCC NO: M2020833.
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