CN116492378A - Application of lactobacillus salivarius LS08 in preparation of lipid-lowering and weight-losing products - Google Patents
Application of lactobacillus salivarius LS08 in preparation of lipid-lowering and weight-losing products Download PDFInfo
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- 241000186869 Lactobacillus salivarius Species 0.000 title claims abstract description 67
- 238000002360 preparation method Methods 0.000 title abstract description 12
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 52
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 26
- 230000004060 metabolic process Effects 0.000 claims abstract description 8
- 230000001603 reducing effect Effects 0.000 claims description 12
- 230000001580 bacterial effect Effects 0.000 claims description 11
- 239000000725 suspension Substances 0.000 claims description 8
- 150000003626 triacylglycerols Chemical class 0.000 claims description 6
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- 150000002632 lipids Chemical class 0.000 abstract description 5
- 238000001727 in vivo Methods 0.000 abstract description 4
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- 239000006041 probiotic Substances 0.000 abstract description 3
- 235000018291 probiotics Nutrition 0.000 abstract description 3
- 208000008589 Obesity Diseases 0.000 abstract description 2
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- 230000000694 effects Effects 0.000 description 16
- 239000013641 positive control Substances 0.000 description 12
- 208000031226 Hyperlipidaemia Diseases 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- OJRHUICOVVSGSY-RXMQYKEDSA-N (2s)-2-chloro-3-methylbutan-1-ol Chemical compound CC(C)[C@H](Cl)CO OJRHUICOVVSGSY-RXMQYKEDSA-N 0.000 description 4
- 229960001770 atorvastatin calcium Drugs 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 210000001325 yolk sac Anatomy 0.000 description 3
- QNVSXXGDAPORNA-UHFFFAOYSA-N Resveratrol Natural products OC1=CC=CC(C=CC=2C=C(O)C(O)=CC=2)=C1 QNVSXXGDAPORNA-UHFFFAOYSA-N 0.000 description 2
- LUKBXSAWLPMMSZ-OWOJBTEDSA-N Trans-resveratrol Chemical compound C1=CC(O)=CC=C1\C=C\C1=CC(O)=CC(O)=C1 LUKBXSAWLPMMSZ-OWOJBTEDSA-N 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
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- VOFUROIFQGPCGE-UHFFFAOYSA-N nile red Chemical compound C1=CC=C2C3=NC4=CC=C(N(CC)CC)C=C4OC3=CC(=O)C2=C1 VOFUROIFQGPCGE-UHFFFAOYSA-N 0.000 description 2
- LUKBXSAWLPMMSZ-UHFFFAOYSA-N resveratrol Chemical compound C1=CC(O)=CC=C1C=CC1=CC(O)=CC(O)=C1 LUKBXSAWLPMMSZ-UHFFFAOYSA-N 0.000 description 2
- 229940016667 resveratrol Drugs 0.000 description 2
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- 201000001320 Atherosclerosis Diseases 0.000 description 1
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- 102000007330 LDL Lipoproteins Human genes 0.000 description 1
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- 210000002249 digestive system Anatomy 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000002008 hemorrhagic effect Effects 0.000 description 1
- 208000020346 hyperlipoproteinemia Diseases 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
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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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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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
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Abstract
The invention discloses application of lactobacillus salivarius LS08 in preparation of a lipid-lowering and weight-losing product, and belongs to the technical field of microorganisms. The lactobacillus salivarius LS08 disclosed by the invention can obviously promote fat metabolism in zebra fish bodies in vivo, can obviously reduce the content of triglyceride and total cholesterol in the hyperlipoidemia zebra fish bodies, has the potential of being applied to weight losing and blood lipid level regulating, and provides theoretical reference and guiding basis for developing a probiotic preparation for treating or preventing obesity and hyperlipoidemia by utilizing the lactobacillus salivarius LS 08.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to application of lactobacillus salivarius LS08 in preparation of lipid-lowering and weight-losing products.
Background
Hyperlipidemia refers to the conditions of low density lipoprotein, glycerate and total cholesterol in a patient, which lead to hyperlipoidemia or hyperlipoproteinemia of the patient, which are not effectively controlled for a long time, and which lead to the occurrence of hyperlipidemia of arteries of the patient, atherosclerosis of the patient, plaque formation easily, arterial lumen stenosis of the patient, and coronary heart disease and ischemic and hemorrhagic cerebral apoplexy. The medicine for treating the hyperlipidemia in the market has the biggest defects of low cure rate, high price, main western medicines and great side effects. Based on the above, screening functional probiotics with cholesterol-reducing effect in human body becomes a research hotspot; aims at screening domestic patent drug strains with strong curative effect and good activity, and has important social value and research significance.
Thus, providing the use of lactobacillus salivarius LS08 in the preparation of products for reducing blood lipid and weight is a problem to be solved by the person skilled in the art.
Disclosure of Invention
In view of this, the invention provides the application of lactobacillus salivarius LS08 in the preparation of products for reducing blood fat and losing weight.
Since zebra fish possess a digestive system similar to humans, such as liver, intestinal tract, etc., digestion and nutrient absorption and transport are highly similar to humans. About 70% of the zebra fish yolk sacs are neutral fat, and if the absorption of the zebra fish yolk sacs can be promoted, the decomposition and metabolism of fat can be promoted, which is similar to the in-vivo fat burning. The yolk sac of zebra fish was stained red by fat-specific fluorescent staining (red), which was clearly observed.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
the application of the lactobacillus salivarius LS08 in preparing the lipid-lowering and weight-losing products is characterized in that the preservation number of the lactobacillus salivarius LS08 (Lactobacillus salivarius) is CGMCC No.22988 (see patent number 202211558776.6).
Further, the lactobacillus salivarius LS08 is applied to the preparation of products for promoting fat metabolism.
Further, the lactobacillus salivarius LS08 is applied to the preparation of products for reducing triglyceride and total cholesterol.
Further, the lactobacillus salivarius LS08 is a bacterial suspension.
At the same concentration, the promotion effect of the lactobacillus salivarius LS08 on the fat metabolism in the zebra fish body is stronger than that of the lactobacillus salivarius 11741, and the lactobacillus salivarius LS08 has good lipid-lowering and weight-losing effects. At the same concentration, the lactobacillus salivarius LS08 has stronger effect of reducing triglyceride and total cholesterol in the zebra fish hyperlipidemia model body than the lactobacillus salivarius 11741, and has good blood lipid reducing effect.
Compared with the prior art, the application of the lactobacillus salivarius LS08 in preparing lipid-lowering and weight-losing products is disclosed, the lactobacillus salivarius LS08 can obviously promote fat metabolism in zebra fish bodies in vivo, can obviously reduce the content of Triglyceride (TG) and Total Cholesterol (TC) in the hyperlipoidemia zebra fish bodies, has the potential of being applied to losing weight and regulating blood lipid level, and provides theoretical reference and guiding basis for developing probiotic preparations for treating or preventing obesity and hyperlipoidemia by using the lactobacillus salivarius LS 08.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.
FIG. 1 is a visual chart showing the effect of Lactobacillus salivarius LS08 on fat levels in zebra fish according to the present invention;
wherein A: normal group; b: a positive control group; c: 1X 10 6 CFU/mL lactobacillus salivarius 11741; d: 1X 10 6 CFU/mL Lactobacillus salivarius LS08;
FIG. 2 is a graph showing the statistical effect of Lactobacillus salivarius LS08 on fat levels in zebra fish according to the present invention;
FIG. 3 is a graph showing the effect of Lactobacillus salivarius LS08 of the present invention on triglycerides in a zebra fish hyperlipidemia model;
FIG. 4 is a graph showing the effect of Lactobacillus salivarius LS08 of the present invention on total cholesterol in a zebra fish hyperlipidemic model.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Resveratrol was purchased from Shanghai source leaf biotechnology limited; nile red was purchased from Sigma, usa; atorvastatin calcium was purchased from Shanghai green sources biotechnology limited; both normal feed and high cholesterol feed were purchased from south-access terlafei feed technology limited; the triglyceride measuring kit and the total cholesterol measuring kit are purchased from all companies of Nanjing established bioengineering research; lactobacillus salivarius 11741 (ATCC 11741) was purchased from beijing Bai's biotech limited.
EXAMPLE 1 preparation of Lactobacillus salivarius LS08 suspension (thallus)
Inoculating lactobacillus salivarius LS08 into MRS liquid culture medium after activating culture, culturing at 37deg.C for 24 hr, and centrifuging at 4deg.C for 10min at 6000r/min to obtain thallus precipitate; after the bacterial cell precipitate is washed twice by PBS, the bacterial cell is resuspended by PBS, and the cell concentration is regulated to be 1 multiplied by 10 6 CFU/mL gave a bacterial suspension (cell).
EXAMPLE 2 preparation of Lactobacillus salivarius 11741 bacterial suspension (thallus)
Inoculating lactobacillus salivarius 11741 into MRS liquid culture medium after activating culture, culturing at 37deg.C for 24 hr, and centrifuging at 4deg.C for 10min at 6000r/min to obtain thallus precipitate; after the bacterial cell precipitate is washed twice by PBS, the bacterial cell is resuspended by PBS, and the cell concentration is regulated to be 1 multiplied by 10 6 CFU/mL gave a bacterial suspension (cell).
Example 3 influence of Lactobacillus salivarius LS08 on fat levels in zebra fish
Healthy wild-type AB-line zebra fish that developed to 3dpf (days post fertilization) were selected and placed in 6-well cell culture plates, 15 strips/well. The experiments set up a normal group, a positive control group (resveratrol), a lactobacillus salivarius 11741 intervention group, and a lactobacillus salivarius LS08 intervention group. PBS was added to the normal group, resveratrol solution (50. Mu. Mol/L) was added to the positive control group, and Lactobacillus salivarius 11741 was added to the intervention group (1X 10) 6 CFU/mL) was added 1X 10 6 CFU/mL Lactobacillus salivarius 11741, lactobacillus salivarius LS08 intervention group (1X 10) 6 CFU/mL) was added 1X 10 6 CFU/mL lactobacillus salivarius LS08, 5mL per well, incubation at 28℃for 24h, and replacement of new solution; after 48h incubation, 10ng/mL of nile red solution was added to each well, 5mL of the solution was incubated at 28℃in the dark for 24h, the zebra fish was washed 3 times with PBS, and the fluorescence intensity in the zebra fish was observed under a fluorescence microscope and recorded by photographing. Quantitative statistical analysis of fluorescence intensity (F) in zebra fish was performed using Image J software. The fat levels in zebra fish were calculated as follows:
SPSS 19.0 software was used to statistically process the data, experimental data were all expressed as x+ -SEM data, analyzed by T-test, compared to normal group: *** P<0.005。
as can be seen from fig. 1 and 2, the intensity of red fluorescence in the zebra fish body reflects the level of fat in the zebra fish body; compared with the normal group, the red fluorescence intensity in the zebra fish of the positive control group (resveratrol) is reduced, which indicates that the fat level in the zebra fish of the positive control group is reduced; meanwhile, compared with a normal group (100.00+/-7.68%), the fat level (46.50 +/-3.39%) of the zebra fish body in the positive control group is obviously reduced (p < 0.005), which indicates that the lipid-lowering experiment in the zebra fish body is effective.
From FIGS. 1 and 2, it can be seen that Lactobacillus salivarius 11741 intervenes in the group (1X 10 6 CFU/mL) red fluorescence intensity in zebra fish is similar to that of normal group; at the same time Lactobacillus salivarius 11741 intervention group (1×10) 6 CFU/mL) of the zebra fish in vivo fat level was 89.47 + -7.09%, and there was no significant difference (P) compared with the normal group (100.00+ -7.68%)>0.05). In addition, the Lactobacillus salivarius LS 08-interfered group (1X 10) 6 CFU/mL) the red fluorescence intensity in zebra fish was reduced, similar to the positive control group; simultaneous Lactobacillus salivarius LS08 intervention group (1X 10) 6 CFU/mL) zebra fish showed a fat level of 54.71 + -4.30% and a significant mean difference (P) compared to the normal group (100.00+ -7.68%)<0.005). Therefore, the results show that at the same concentration, the promotion effect of the lactobacillus salivarius LS08 on the fat metabolism in the zebra fish body is stronger than that of the lactobacillus salivarius 11741, and the effects of reducing blood fat and losing weight are good.
Example 4 Effect of Lactobacillus salivarius LS08 on Triglycerides and Total cholesterol in a zebra fish hyperlipidemic model
Model construction: the normal wild type AB-series zebra fish (5 dpf) is selected under microscope and divided into a normal group, a model group (high cholesterol), a positive control group (atorvastatin calcium) and a lactobacillus salivarius 11741 intervention group (1×10) 6 CFU/mL), lactobacillus salivarius LS08 intervention group (1X 10) 6 CFU/mL). Each group is provided with 6 compound holes, 30 fishes are arranged in each hole, 5mL PBS is arranged in each hole, and the zebra fishes in the normal group are fed with common feed (20 mg each time); zebra fish in the model group, the positive control group and the strain intervention group are fed with high cholesterol feed (20 mg each time) 3 times a day, the feeding period is 7 days, and PBS is replaced once a day.
And (3) intervention: adding PBS solution into the normal group and the model group; the positive control group was added with 0.25. Mu.g/mL atorvastatin calcium solution; lactobacillus salivarius 11741 intervention group (1×10) 6 CFU/mL) was added 1X 10 6 CFU/mL lactobacillus salivarius 11741; lactobacillus salivarius LS08 intervention group (1×10) 6 CFU/mL) was added 1X 10 6 CFU/mL lactobacillus salivarius LS08, 5mL per well; meanwhile, the zebra fish of the normal group is fed with common feed (20 mg each time); zebra fish in the model group, positive control group and strain intervention group are fed with high cholesterol feed (20 mg each time) 3 times per day, the feeding period is 5 days, and one time per day is replacedAnd (3) secondary solution. After the intervention is finished, zebra fish are collected to a 1.5mL centrifuge tube, and 6 tubes are collected for each experimental group; after the water in the centrifuge tube was sucked dry, the water was mixed with the following components in mass (mg): volume (μl) is 1:9 adding PBS buffer solution, holding the micro-electric tissue homogenizer with S-18KS to break the zebra fish homogenate until no obvious tissue fragments are present, centrifuging at 15000 Xg at 4deg.C for 15min, and collecting supernatant. Triglyceride and total cholesterol concentrations of each group were determined using a triglyceride determination kit and a total cholesterol determination kit.
SPSS 19.0 software was used to statistically process the data, experimental data were all expressed as x+ -SEM data, analyzed by T-test, compared to normal group: ### P<0.05; compared with the model group: ** P<0.01， *** P<0.005。
as can be seen from FIGS. 3 and 4, compared with the normal group (triglyceride: 0.09.+ -. 0.01mmol/gprot, total cholesterol: 0.07.+ -. 0.01 mmol/gprot), the triglyceride and total cholesterol (triglyceride: 0.32.+ -. 0.02mmol/gprot, total cholesterol: 0.29.+ -. 0.02 mmol/gprot) in the zebra fish of the model group are both significantly increased (P < 0.005), indicating that the current zebra fish hyperlipidemia model establishment was successful.
As can be seen from FIGS. 3 and 4, the triglyceride and total cholesterol of the zebra fish in the positive control group were 0.15.+ -. 0.01mmol/gprot and 0.16.+ -. 0.01mmol/gprot, respectively, and the differences were significant (P < 0.005) compared with the model group (triglyceride: 0.32.+ -. 0.02mmol/gprot, total cholesterol: 0.29.+ -. 0.02 mmol/gprot). Therefore, the atorvastatin calcium has the effect of reducing blood fat, and is consistent with clinical results, so that the blood fat reducing efficacy evaluation test is effective.
Lactobacillus salivarius 11741 intervention group (1×10) 6 CFU/mL) triglycerides and total cholesterol in zebra fish were 0.26±0.03mmol/gprot and 0.27±0.01mmol/gprot, respectively, and compared with the model group (triglyceride: 0.32.+ -. 0.02mmol/gprot, total cholesterol: 0.29.+ -. 0.02 mmol/gprot) compared with no significant difference (P>0.05). In addition, lactobacillus salivarius LS08 intervention group (1×10 6 CFU/mL) triglycerides and total cholesterol in zebra fish were 0.17±0.01mmol/gprot and 0.18±0.01mmol/gprot, respectively, and compared with the model group (triglyceride: 0.32.+ -. 0.02mmol/gprot, total cholesterol: 0.29.+ -. 0.02 mmol/gprot)Significant difference in phase (P<0.01). Therefore, the results show that at the same concentration, the lactobacillus salivarius LS08 has stronger effect of reducing triglyceride and total cholesterol in the zebra fish hyperlipidemia model body than the lactobacillus salivarius 11741, and has good blood lipid reducing effect.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (6)
1. The application of the lactobacillus salivarius LS08 in preparing the lipid-lowering and weight-losing products is characterized in that the preservation number of the lactobacillus salivarius LS08 is CGMCC No.22988.
2. Use of lactobacillus salivarius LS08 in the manufacture of a lipid-lowering and weight-losing product according to claim 1, wherein said lactobacillus salivarius LS08 is a bacterial suspension.
3. Use of lactobacillus salivarius LS08 as claimed in claim 1 in the manufacture of a product for promoting fat metabolism.
4. Use of lactobacillus salivarius LS08 in the manufacture of a product for promoting fat metabolism according to claim 3 wherein said lactobacillus salivarius LS08 is a bacterial suspension.
5. Use of lactobacillus salivarius LS08 as claimed in claim 1 in the manufacture of a product for reducing triglycerides and total cholesterol.
6. Use of lactobacillus salivarius LS08 according to claim 5 for the manufacture of products for reducing triglycerides and total cholesterol, wherein said lactobacillus salivarius LS08 is a bacterial suspension.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117384788A (en) * | 2023-10-10 | 2024-01-12 | 广东悦创生物科技有限公司 | Saliva combined lactobacillus SM4 and application thereof in preparation of whitening and cholesterol lowering foods and medicines |
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|---|---|---|---|---|
| CN117384788A (en) * | 2023-10-10 | 2024-01-12 | 广东悦创生物科技有限公司 | Saliva combined lactobacillus SM4 and application thereof in preparation of whitening and cholesterol lowering foods and medicines |
| CN117384788B (en) * | 2023-10-10 | 2024-03-22 | 广东悦创生物科技有限公司 | Saliva combined lactobacillus SM4 and application thereof in preparation of whitening and cholesterol lowering foods and medicines |
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