CN118028156A - Brevibacillus laterosporus with blood glucose and lipid reducing effects and application thereof - Google Patents
Brevibacillus laterosporus with blood glucose and lipid reducing effects and application thereof Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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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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- 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
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- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention discloses a Brevibacillus laterosporus strain with the effects of reducing blood sugar and lipid and application thereof. The strain is Brevibacillus laterosporus (Brevibacillus laterosporus) ASD0001, and the preservation number is GDMCC No:63890. The Brevibacillus laterosporus is autonomously screened from soil, has the effects of reducing the weight of obese animals, inhibiting adipose tissue formation and improving glucose tolerance and insulin resistance of the obese animals, can be applied to preparing products with functions of reducing blood sugar and lipid or products for preventing and treating diabetes, and has good market application prospects.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to Brevibacillus laterosporus with the effects of reducing blood sugar and lipid and application thereof.
Background
With the improvement of the living standard of people, the proportion of obese people is higher and higher, and obesity has become one of the most serious public health problems in the global scope. The world obesity Association has recently issued a global obesity map of 2023 showing that by 2035, over 40 million people worldwide will be obese or overweight, accounting for 51% of the general population. The obesity causes various reasons, besides the well-known problems of diet structure, emotion, exercise frequency, living habit and the like, and the obesity problem is difficult to be regulated by singly depending on diet and is difficult to be sustained. However, obesity is not only related to the problem of visual appearance, but also important for human health, for example, obesity is often accompanied with diseases of heart and cerebral vessels, liver, kidney and other parts, so how to control the occurrence of obesity is more and more important for human health.
In addition to diet, exercise, regular life, etc., weight loss by medication is a relatively easy way to control obesity. Several drugs are commonly known, one being digestive enzyme inhibitors such as orlistat, which, by binding to pancreatic lipase in the intestine, prevent the breakdown and absorption of glycerol and fat in the food; the other is an appetite suppressant, such as dexfenfluramine, fenfluramine, and the like, which is a drug that suppresses appetite by stimulating neurotransmitter production, release and inhibiting its reuptake, essentially by reducing food intake. However, taking these chemically synthesized weight-reducing drugs may cause serious side effects, such as dry mouth, constipation, dizziness, insomnia, etc., and the national food and drug administration has stopped the production, sales and use of sibutramine preparations and raw materials in China since 2010. As another example, orlistat can cause adverse gastrointestinal reactions including flatulence, stool urgency, increased stool frequency, and lipidic stool. The semaglutin has high attention in the weight-losing market, can realize two effects of reducing blood sugar and weight, and has the main mechanism that the semaglutin can promote insulin secretion in a glucose concentration dependent manner by activating GLP-1 (glucagon-like peptide-1) receptor, inhibit glucagon secretion, delay gastric emptying to increase satiety and inhibit appetite by inhibiting the action of hypothalamic feeding center, so that the effect of reducing blood sugar and weight is achieved, but the semaglutin also has certain side effects.
Therefore, developing a weight-reducing product with little side effect and good effect is a category of current market shortage.
Disclosure of Invention
The first object of the invention is to provide a Brevibacillus laterosporus (Brevibacillus laterosporus) ASD0001 strain with the effects of reducing blood sugar and lipid. The strain was deposited at the cantonese microbiological strain collection center (GDMCC), address: 30, building 5, post code of 30 # 100 college in Guangzhou city View area martyr: 510070, the preservation number is GDMCCNo:63890, and the preservation date is 2023, 10, 19.
The second purpose of the invention is to provide the application of the Brevibacillus laterosporus ASD0001 in preparing products with the functions of reducing blood sugar and lipid.
Preferably, the active ingredient of the functional product for reducing blood glucose and lipid comprises thallus of Brevibacillus laterosporus ASD0001 or a fermentation product thereof.
Preferably, the functional product for reducing blood sugar and lipid comprises food, health care products or medicines.
The third object of the invention is to provide the application of the Brevibacillus laterosporus ASD0001 in preparing products for preventing and treating diabetes.
Preferably, the product for preventing and treating diabetes comprises health care products or medicines.
The fourth object of the present invention is to provide a functional product for lowering blood sugar and reducing blood lipid, which comprises as an active ingredient a bacterial cell of Brevibacillus laterosporus ASD0001 or a fermentation product thereof.
A fifth object of the present invention is to provide a product for preventing and treating diabetes, which comprises as an active ingredient a bacterial cell of Brevibacillus laterosporus ASD0001 or a fermentation product thereof.
The invention has the following beneficial effects:
The Brevibacillus laterosporus is autonomously screened from soil, has the effects of reducing the weight of obese animals, inhibiting adipose tissue formation and improving glucose tolerance and insulin resistance of the obese animals, can be applied to preparing products with functions of reducing blood sugar and lipid or products for preventing and treating diabetes, and has good market application prospects.
Brevibacillus laterosporus (Brevibacillus laterosporus) ASD0001 was deposited at the microorganism strain collection (GDMCC) of Guangdong province, 5 th floor of the No. 100 college of Guangzhou city martyr, post code: 510070, accession number is: GDMCC No:63890.
Drawings
FIG. 1 shows the weight difference between BL1 and ASD0001 mice after 4 weeks of high fat diet feeding. A: the body shape comparison of the mice in the 6 th four groups is that: control group (C), model group (H), BL1 group (BL 1), ASD0001 group (ASD 0001); b: four groups of mice were assayed for 6 weeks of body weight. * Representing the significance of differences in body weight of ASD0001 group versus model group and BL1 group, * represents p < 0.05, ** represents p < 0.01.
FIG. 2 shows the weight difference of Inguinal White Adipose Tissue (iWAT) of BL1 and ASD0001 mice after 4 weeks of high-fat diet feeding. A: four groups of mice iWAT were photographed, in order from left to right: control group (C), model group (H), BL1 group (BL 1), ASD0001 group (ASD 0001); b: four groups of mice iWAT were assayed by weight. ## Representing that the difference between the group H and the group C is extremely significant p < 0.01, ** represents that the difference between ASD0001 and the group H and the group BL1 is extremely significant p < 0.01.
FIG. 3 shows the epididymal white adipose tissue (eWAT) weight difference between BL1 and ASD0001 mice after 4 weeks of high-fat diet feeding. A: four groups of mice were photographed with eWAT, in order from left to right: control group (C), model group (H), BL1 group (BL 1), ASD0001 group (ASD 0001); b: four groups of mice were weighed. ## Representing that the difference between the group H and the group C is extremely significant p < 0.01, ** represents that the difference between ASD0001 and the group H is extremely significant p < 0.01, and * represents that the difference between ASD0001 and the group BL1 is significantly p < 0.05.
FIG. 4 shows the results of oil red O staining of liver tissue of BL1 and ASD0001 mice after 4 weeks of high-fat diet feeding. In the figure, C: control group, H: model group, BL1: BL1 treatment group, ASD0001: ASD0001 treatment group.
FIG. 5 shows the glucose tolerance differences between BL1 and ASD0001 mice after 4 weeks of high fat diet feeding. In the figure, C: control group, H: model group, BL1: BL1 treatment group, ASD0001: ASD0001 treated group ## represents very significant p < 0.01 for group H compared to group C, ** represents very significant p < 0.01 for ASD0001 compared to group H.
FIG. 6 shows the difference in insulin resistance between BL1 and ASD0001 mice after 4 weeks of high-fat diet feeding. In the figure, C: control group, H: model group, BL1: BL1 treatment group, ASD0001: ASD0001 treated group ## represents very significant p < 0.01 in comparison to group C, * represents significant p < 0.05 in comparison to group H.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
The following examples relate to strain Brevibacillus laterosporus BL1, deposited under the accession number GDMCC No:62699, which is disclosed in the literature Weng GY,Huang J,et al.Brevibacillus laterosporus BL1,a promising probiotic,prevents obesity and modulates gut microbiota in mice fed a high-fat diet.Frontiers in Nutrition.2022Nov 24:9:1050025.
Example 1: lipid lowering effect of Brevibacillus laterosporus ASD0001
40 Male C57BL/6 mice (purchased from the Betty Biotechnology Co., ltd.) of 5 weeks old were individually kept in a controlled environment (24.+ -. 2 ℃ C., 45-60% relative humidity, 12h/12h light and dark cycle) and were free to eat during the whole experiment. After 1 week of adaptation, the mice were randomly divided into 4 groups (n=10 per group), respectively, control group (C), model group (H), BL1 group (BL 1), ASD0001 group (ASD 0001), wherein the mice of the control group were fed normal diet (cat No. XTCON50 0J, available from the company limited), while the mice of the model group, BL1 group, and ASD0001 group were fed high fat diet (cat No. XTHF60, available from the company limited), and after 4 weeks of high fat diet, the mice of the BL1 group and ASD0001 group were respectively perfused with the strain BL1 (1×10 9 CFU/d) and ASD0001 (1×10 9 CFU/d) for 6 weeks, and the mice of the control group and model group were perfused with an equal volume of normal saline. Body Weight (BW) of mice was recorded weekly. At the end of 6 weeks of treatment, the mice body weight was determined. After mice were sacrificed, the liver, inguinal White Adipose Tissue (iWAT) and epididymal white adipose tissue (eWAT) were taken, weights of iWAT and eWAT were measured, and the liver was fixed and then oil red O staining was performed to observe the adipogenesis level of the liver tissue, and the results are shown in FIGS. 1 to 4.
Figure 1 shows that after 4 weeks of high fat diet feeding mice (week 0 for weight calculation), the weight was significantly higher than the control group C mice. BL1 and ASD0001 were fed for 6 weeks, and model group H mice were significantly obese compared to control group C mice. BL1 and ASD0001 mice were significantly leaner than model group H (FIG. 1A). The body weight of each group of mice was measured weekly, and from the results, BL1 group and ASD0001 group were lower than model group H in each of the groups after 1-6 weeks of feeding with the bacterial liquid. After 5 and 6 weeks of feeding, ASD0001 group mice had significantly lower body weight than model group H and significantly lower body weight than BL1 group. It was demonstrated that ASD0001 was more effective in controlling body weight of obese mice than strain BL 1.
FIG. 2 shows that after 4 weeks of high fat diet feeding mice, BL1 and ASD0001 were fed for 6 weeks, each group of mice was dissected and iWAT weight was measured. From the results, iWAT weight of model group H was significantly higher than control group C, iWAT weight of the ASD0001 and BL1 bacteria fed mice was significantly lower than model group H, and iWAT weight of ASD0001 was significantly lower than BL 1. It was demonstrated that ASD0001 was more effective in reducing the formation of mice iWAT compared to strain BL 1.
FIG. 3 shows that after 4 weeks of high fat diet feeding mice, BL1 and ASD0001 were fed for 6 weeks, each group of mice was dissected and eWAT weights were determined. From the results, the weight of the eWAT was significantly higher in model group H than in control group C, the weight of the eWAT was significantly lower in ASD 0001-and BL 1-fed mice than in model group H, and the weight of the eWAT was significantly lower in ASD0001 group than in BL1 group. It was demonstrated that ASD0001 was more effective in reducing the formation of mouse eWAT compared to strain BL 1.
FIG. 4 shows that after 4 weeks of high fat diet feeding mice, BL1 and ASD0001 were infused for 6 weeks, liver tissue was dissected from each group of mice, and the adipogenesis level of liver tissue was compared in four groups of mice. As can be seen, the adipogenesis level of model group H was significantly higher than that of control group C. The ASD0001 group and BL1 bacteria group can effectively reduce the adipogenesis of mice. It was demonstrated that ASD0001 was more effective in reducing fat formation in the liver than strain BL 1.
Example 2: hypoglycemic effect of Brevibacillus laterosporus ASD0001
40 Male C57BL/6 mice of 5 weeks old were individually kept in a controlled environment (24.+ -. 2 ℃ C., 45-60% relative humidity, 12h/12h light and dark cycle) and were free to eat throughout the experiment. After 1 week of adaptation, the mice were randomly divided into 4 groups (n=10 per group), respectively, control group (C), model group (H), BL1 group (BL 1), ASD0001 group (ASD 0001), wherein the mice of the control group were fed normal diet (cat No. XTCON50 0J, available from the company limited), while the mice of the model group, BL1 group, and ASD0001 group were fed high fat diet (cat No. XTHF60, available from the company limited), and after 4 weeks of high fat diet, the mice of the BL1 group and ASD0001 group were respectively perfused with the strain BL1 (1×10 9 CFU/d) and ASD0001 (1×10 9 CFU/d) for 6 weeks, and the mice of the control group and model group were perfused with an equal volume of normal saline. Abdominal glucose tolerance test (IGTT) and insulin resistance test (ITT) were performed at week 7. Tail vein blood glucose levels were measured using a glucometer. IGTT analysis, mice were intraperitoneally injected with glucose (2 g/kg body weight) after overnight fast, and then blood glucose levels were measured at injection 0, 15, 30, 60, 90, and 120 min. ITT analysis, mice were intraperitoneally injected with insulin (0.75U/kg body weight) 6h after fasting, and then blood glucose levels at 0, 15, 30, 60, 90, and 120min were measured. The results are shown in FIGS. 5-6.
Figure 5 shows that ASD0001 and BL1 were fed 6 weeks after 4 weeks of high fat diet feeding to mice. Mice fasted overnight were intraperitoneally injected with glucose (2 g/kg body weight) and blood glucose levels were measured at 0, 15, 30, 60, 90 and 120min, respectively, with model group H glucose levels on average significantly higher than control group C. After 30, 60, 90 and 120min post glucose injection, the blood glucose levels were significantly lower in ASD0001 and BL1 groups than in model group H. For high-fat induced obese mice, ASD0001 was more effective in improving glucose tolerance in mice than strain BL1.
Figure 6 shows that ASD0001 and BL1 were fed 6 weeks after 4 weeks of high fat diet feeding to mice. After 6h of fasted mice, insulin (0.75U/kg body weight) was intraperitoneally injected and blood glucose levels were measured at 0, 15, 30, 60, 90 and 120min, respectively. Model group H was higher than control group C at each time. The blood glucose levels were lower in the asd0001 group than in the model group H and BL1 groups at 15min and 120min after insulin injection. For fat-induced obese mice, ASD0001 was more effective in improving insulin resistance in mice than strain BL1.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that the above-mentioned preferred embodiment should not be construed as limiting the invention, and the scope of the invention should be defined by the appended claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and such modifications and adaptations are intended to be comprehended within the scope of the invention.
Claims (8)
1. Brevibacillus laterosporus (Brevibacillus laterosporus) ASD0001, accession number: GDMCC No:63890.
2. The use of Brevibacillus laterosporus ASD0001 in preparing a hypoglycemic and hypolipidemic functional product.
3. The use according to claim 2, wherein the active ingredient of the functional product for reducing blood glucose and lipid comprises the thallus of Brevibacillus laterosporus ASD0001 or the fermentation product thereof.
4. The use according to claim 2, wherein the functional product for reducing blood glucose and blood lipid comprises a food, a health product or a pharmaceutical product.
5. Use of Brevibacillus laterosporus ASD0001 in the preparation of a product for preventing and treating diabetes.
6. The use according to claim 5, wherein the product for preventing and treating diabetes comprises a health product or a pharmaceutical product.
7. A functional product for reducing blood glucose and lipid comprising the Bacillus laterosporus ASD0001 or a fermentation product thereof according to claim 1 as an active ingredient.
8. A product for preventing and treating diabetes, comprising the Brevibacillus laterosporus ASD0001 or a fermentation product thereof as an active ingredient.
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