Lactobacillus strain for preventing re-fatting as well as product, composition and application thereof

Abstract

The invention provides a lactobacillus strain as well as a product, a composition and application thereof. In a first aspect of the present disclosure, there is provided a strain of the family Lactobacillus, which can include a strain MNH08891 having a preservation number of GDMCC NO. 65551, or a strain MNHH22076 having a preservation number of GDMCC NO. 65554, or a strain having at least 98.65% identity with the strain MNH08891 or a 16S rRNA of the strain MNHH22076, or a strain having at least 98.65% identity with the 16S rRNA sequence set forth in SEQ ID NO: 1. The strain disclosed by the embodiment of the invention or a product based on the strain has good effects of treating, preventing or relieving metabolic diseases and treating, preventing or improving metabolic disorders after drug withdrawal for treating the metabolic diseases.

Description

Technical Field

[0001] The present disclosure generally relates to the field of microbial technology. More specifically, the present disclosure relates to a strain of Lactobacillus family for preventing obesity, and its products, compositions and applications. Background Art

[0002] Obesity has become an increasingly serious public health problem worldwide. Faced with this challenge, the medical community is committed to developing various treatments, one of which is based on incretin-like peptide-1 (GLP-1) receptor agonists. There is growing evidence that patients' weight can be significantly reduced by drugs that mimic insulin secretion. These drugs help suppress appetite and reduce food intake. Although they have achieved significant results in weight loss, a large number of patients still stop using them for various reasons during actual use.

[0003] Although the weight loss effect of GLP-1 receptor agonists such as liraglutide and semaglutide is indeed significant in the short term, studies have found that the incidence of four gastrointestinal adverse reactions induced by GLP-1 agonists semaglutide and liraglutide is higher than other weight loss drugs, including biliary tract disease, pancreatitis, intestinal obstruction and gastric spasm. In addition, GLP-1 receptor agonists are prone to rebound after discontinuation of medication. Clinical trials have shown that participants who received semaglutide treatment recovered more than 50% of their lost weight one year after discontinuation of the drug. In other words, persistent use of semaglutide can indeed ensure weight loss; but once the intervention stops, the possibility of rebound may be even higher.

[0004] Studies in the population found that from enrollment to one year after discontinuation, the liraglutide group rebounded a total of 8.7 kg, the most among all intervention groups, while the combined intervention group of liraglutide + moderate to high intensity exercise had the least weight rebound, 5.1 kg less than the liraglutide group (P=0.04). Researchers believe that one of the reasons why liraglutide is easy to regain weight after discontinuation of liraglutide is that liraglutide achieves weight loss by suppressing appetite and delaying gastric emptying, which is equivalent to forcing us to control our mouths. However, once we stop taking the medicine, we may not be able to cope with the restored appetite, which eventually leads to weight rebound. (Jensen SBK, Blond MB, Sandsdal RM, et al. Healthy weight loss maintenance with exercise, GLP-1 receptor agonist, or both combined followed by one year without treatment: a post-treatment analysis of a randomised placebo-controlled trial[J]. eClinicalMedicine, 2024.) That is to say, due to human intervention, that is, the increase of exogenous GLP-1, the body detects a higher level of "fake" GLP-1, and the synthesized endogenous GLP-1 will decrease accordingly. After the drug is suddenly stopped, the originally high level of GLP-1 in the body drops suddenly, hunger and appetite will "return with a vengeance", leading to a rapid rebound in weight, blood sugar, blood lipids, and body fat.

[0005] It can be seen that most of the existing weight loss products on the market only treat the symptoms and not the root cause. Some even affect physical health, destroy the balance of intestinal flora, disrupt metabolism, easily cause rebound or dependence, and even cause serious side effects. Most people's weight will return to the previous level after stopping the drug, and health indicators such as blood pressure, blood sugar and cholesterol levels will also deteriorate. Due to the significant problem of weight rebound after stopping the drug, how to inhibit weight regain and maintain blood sugar balance after stopping GLP-1 agonists has become an urgent problem to be solved.

[0006] Probiotics have shown potential in treating metabolic disorders, including obesity, diabetes, and metabolic-associated fatty liver disease (MAFLD). However, since appetite will "return with a vengeance" after discontinuation of weight-loss drugs, making weight loss more difficult, screening out new species or strains from probiotics that can be effectively used to prevent weight regain remains a huge challenge, but also represents a huge unmet need. Summary of the invention

[0007] In order to at least solve one or more of the technical problems mentioned above, the present disclosure provides a lactobacillus and its products, compositions and uses in multiple aspects.

[0008] The present invention discloses the isolation of a new Lactobacillus family ( Lactobacillaceae ) strains MNH08891 and MNH22076.

[0009] The strain disclosed in the present invention can effectively prevent weight regain and maintain healthy weight, blood sugar and body fat after weight loss. Specifically, the effects of preventing weight regain of the strain disclosed in the present invention include: 1. improving weight regain after drug withdrawal and maintaining weight after weight loss; 2. improving blood sugar homeostasis imbalance after drug withdrawal and maintaining blood sugar balance; 3. improving local fat accumulation after drug withdrawal and maintaining local fat weight loss effect.

[0010] The new strain disclosed in the present invention is tolerant to salt and choline and is easy to colonize; it can produce acid using a variety of carbon sources and is easy to culture; it is sensitive to most types of antibiotics and has a low risk of antibiotic resistance; it is safe and non-toxic. It can reduce weight gain and weight gain rate after discontinuation of semaglutide treatment; improve the imbalance of blood sugar homeostasis after discontinuation of semaglutide treatment and improve blood sugar elevation; significantly reduce fat accumulation after discontinuation of semaglutide treatment, and has the purpose of preventing blood sugar elevation and fat accumulation in the body, preventing weight gain again, and maintaining blood sugar balance.

[0011] According to a first aspect of the present disclosure, the present disclosure provides an isolated Lactobacillus family ( Lactobacillaceae ) strain, the average nucleotide identity (ANI) values ​​of which are 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97%, 97.1%, 97.2%, 97.9%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 98.1%, 98.2%, 98.9%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 98.1%, 98.2%, 98. 7.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%.

[0012] In some embodiments, the Lactobacillaceae ( Lactobacillaceae) strain with a deposit number of GDMCC No: 65551 or a deposit number of GDMCC No: 65554. The alignment score (AF) is at least 90%, at least 90.5%, at least 91.5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.5%, at least 96%, at least 96.5%, at least 97%, at least 97.5%, at least 98%, at least 98.5%, at least 99%, at least 99.5% or 100%.

[0013] In some embodiments, the Lactobacillaceae ( Lactobacillaceae ) strain and the strain with a deposit number of GDMCC No: 65551 or a deposit number of GDMCC No: 65554 have a 16S rRNA sequence that is at least 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.65%, at least 98.7%, at least 98.8%, at least 98.9%, at least 99.0%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9% or 100% identical to the sequence shown in SEQ ID NO.1 or 2.

[0014] In some embodiments, the lactobacillus is Lactobacillus paracasei ( Lacticaseibacillus paracasei )MNH08891, deposited in Guangdong Microbiological Culture Collection Center (GDMCC), the deposit number is GDMCC No: 65551, and the deposit date is November 28, 2024.

[0015] In some embodiments, the lactobacillus is Lactobacillus crispatus ( Lactobacillus crispatus )MNH22076, deposited in Guangdong Microbiological Culture Collection Center (GDMCC), with the deposit number GDMCC No: 65554 and the deposit date of November 28, 2024.

[0016] In some embodiments, the Lactobacillus paracasei MNH08891 has a gene cluster that produces a primary metabolite that converts pyruvate into acetate and formate.

[0017] In some embodiments, the Lactobacillus crispatus MNH22076 has a gene cluster that can produce a primary metabolite that converts arginine into bicarbonate.

[0018] According to a second aspect of the present disclosure, provided is a culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillusaceae strain described in the first aspect.

[0019] In some embodiments, the culture includes a fermentation broth of the Lactobacillusaceae strain or a concentrated or dried product of the fermentation broth.

[0020] In some embodiments, the culture comprises a solid culture, a fermentation culture, or a supernatant of a fermentation culture of a Lactobacillusaceae strain.

[0021] In some embodiments, the fermentation culture or fermentation culture supernatant is a fermentation culture or fermentation culture supernatant obtained under anaerobic culture conditions using a liquid culture medium.

[0022] According to a third aspect of the present disclosure, a composition for preventing obesity recurrence is provided, comprising an effective amount of a culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillus strain of the first aspect or the Lactobacillus strain of the second aspect; wherein the culture comprises any one of the following A) to D): A) fermentation broth of the strain; B) the supernatant of the fermentation broth of the strain; C) an inactivated fermentation broth of the strain; D) The concentrated or dried product of any one of A) to C) above.

[0023] In some embodiments, the composition is provided in liquid form or solid form.

[0024] In some embodiments, the composition comprises 1×10 4 Up to 1×10 12 cfu / mL or 1×10 4 Up to 1×10 12 cfu / mg of the live Lactobacillus bacteria.

[0025] In some embodiments, the composition comprises 1×10 5 Up to 1×10 11 cfu / mL or 1×10 5 Up to 1×10 11 cfu / mg of the live Lactobacillus bacteria.

[0026] In some embodiments, the composition comprises 1×10 6 Up to 1×10 10 cfu / mL or 1×10 6 Up to 1×10 10 cfu / mg of the live Lactobacillus bacteria.

[0027] In some embodiments, the composition comprises 1×10 7 Up to 1×10 9 cfu / mL or 1×10 7Up to 1×10 9 cfu / mg of the live Lactobacillus bacteria.

[0028] In some embodiments, each gram of the composition contains 1×10 3 Up to 1×10 17 colony forming units (CFU) of bacteria; for example, 1×10 4 Up to 1×10 12 pcs, 1×10 5 Up to 1×10 11 or 1×10 6 Up to 1×10 10 Colony forming units (CFU), for example, 1×10 3 , 2×10 3 , 3×10 3 , 4×10 3 , 5×10 3 , 6×10 3 ,7×10 3 , 8×10 3 ,9×10 3 , 1×10 4 , 2×10 4 , 3×10 4 , 4×10 4 , 5×10 4 , 6×10 4 ,7×10 4 , 8×10 4 ,9×10 4 , 1×10 5 , 2×10 5 , 3×10 5 , 4×10 5 , 5×10 5 , 6×10 5 ,7×10 5 , 8×10 5 ,9×10 5 , 1×10 6 , 2×10 6 , 3×10 6 , 4×10 6 , 5×10 6 , 6×10 6 ,7×10 6 , 8×10 6 ,9×10 6 , 1×10 7 , 2×10 7 , 3×10 7 , 4×10 7 , 5×10 7、6×10 7 、7×10 7 、8×10 7 、9×10 7 、1×10 8 、2×10 8 、3×10 8 、4×10 8 、5×10 8 、6×10 8 、7×10 8 、8×10 8 、9×10 8 、1×10 9 、2×10 9 、3×10 9 、4×10 9 、5×10 9 、6×10 9 、7×10 9 、8×10 9 、9×10 9 、1×10 10 、2×10 10 、3×10 10 、4×10 10 、5×10 10 、6×10 10 、7×10 10 、8×10 10 、9×10 10 、1×10 11 、2×10 11 、3×10 11 、4×10 11 、5×10 11 、6×10 11 、7×10 11 、8×10 11 、9×10 11 、1×10 12 、2×10 12 、3×10 12 、4×10 12 、5×10 12 、6×10 12 、7×10 12 、8×10 12 、9×10 12 、1×10 13 、2×10 13 、3×10 13 、4×10 13 、5×10 13 、6×10 13 、7×1013 , 8×10 13 ,9×10 13 In some embodiments, the lactobacillus in the composition is an attenuated bacterium, a killed bacterium, a freeze-dried bacterium, or an irradiated bacterium, for example, a heat-killed bacterium, preferably a pasteurized bacterium.

[0029] In some embodiments, the composition is in the form of a liquid, foam, cream, spray, powder (eg, lyophilized powder), or gel.

[0030] In some embodiments, the composition is in the form of powder, microencapsulated powder, capsule, tablet, lozenge, granule, oral solution, suspension, emulsion, liquid preparation, sustained release preparation, nanoformulation or microencapsulated capsule.

[0031] In some embodiments, the composition is in the form of an oral dosage or an injection.

[0032] In some embodiments, the composition further comprises one or more pharmaceutically acceptable carriers, food carriers, active agents, excipients and / or adjuvants.

[0033] In some embodiments, the pharmaceutically acceptable excipients are well known to those skilled in the art.

[0034] In some embodiments, the auxiliary material may be at least one selected from a carrier, an excipient, a diluent, a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, and a flavor.

[0035] In some embodiments, the composition comprises one or more of a buffer (e.g., sodium bicarbonate, infant formula or sterilized human milk or other agents that allow bacteria to survive and grow (e.g., survive in the acidic environment of the stomach and grow in the intestinal environment)), a lyoprotectant, a preservative, a stabilizer, a binder, a compacting agent, a lubricant, a dispersion enhancer, a disintegrant, an antioxidant, a flavoring agent, a sweetener, and a coloring agent.

[0036] In some embodiments, the active agent also includes one or more agents for preventing or treating cardiovascular and cerebrovascular diseases, metabolic diseases, lowering lipids, and improving intestinal flora.

[0037] The active agent has at least one of the following functions: (a) suppressing appetite, (b) preventing metabolic diseases, and (c) treating metabolic diseases.

[0038] In some embodiments, the active agent is selected from: a GLP-1 receptor agonist, a dual agonist of a GLP-1 receptor and a GCG receptor, a triple agonist of a GLP-1 receptor, a GIP receptor and a GCG receptor, an AMPK agonist or an active drug that promotes GLP-1 secretion.

[0039] In some embodiments, the active agent is selected from metformin, sulfonylureas, meglitinide, thiazolidinediones, DPP-4 inhibitors, GLP-1 receptor agonists, SGLT2 inhibitors, insulin, pioglitazone, rosiglitazone, pentoxifylline, omega-3-fatty acids, statins, ezetimibe, and ursodeoxycholic acid.

[0040] In some embodiments, the GLP-1 receptor agonist includes one or more of the following: Exenatide, Liraglutide, Lixisenatide, Dulaglutide, Albiglutide, Semaglutide, Betaglutide, Losenapeptide, and Somalutide.

[0041] In some embodiments, the active agent is one or more of probiotics, prebiotics, or a combination thereof.

[0042] In some embodiments, the probiotics are selected from one or more of lactic acid bacteria, lactobacillus, lactococci, butyrate-producing bacteria, bifidobacteria, thermophilic Streptococcus, faecal Streptococcus, and mesenteric Leuconostoc.

[0043] In some embodiments, the prebiotic is selected from the group consisting of inulin, mulberry leaf extract, berberine, ganoderma lucidum, green coffee bean extract, oats, pectin, potato or its extract, citrus polyphenols, Ceylon cinnamon, chromium, ergothioneine, astaxanthin, quercetin, curcumin, proanthocyanidins, resistant dextrin, yeast β-glucan, ginseng or its extract, nutritional compounds, biotin, polydextrose, fructooligosaccharide (FOS), galacto-oligosaccharide (GOS), starch, cellulose, b-glucan, hemicellulose, lactulose, manno-oligosaccharide, manno-oligosaccharide (MOS), inulin rich in fructooligosaccharide, oligo-glucose, tagatose, trans-galacto-oligosaccharide, pectin, resistant starch, xylo-oligosaccharide (XOS) and any combination thereof.

[0044] In some embodiments, the composition can be formulated as a frozen composition, such as a frozen composition prepared by quick freezing and drying, or lyophilization, for storage and / or transportation.

[0045] In some embodiments, the composition is obtained by spray drying. In some embodiments, the composition is obtained by electrostatic spray drying.

[0046] In some embodiments, the strain in the composition is freeze-dried or spray-dried. In some embodiments, the strain in the composition is electrostatically spray-dried. In some embodiments, the strain in the composition is freeze-dried or spray-dried and is alive. In some embodiments, the strain in the composition is freeze-dried or spray-dried and is capable of partially or completely colonizing the intestine. In some embodiments, the strain is reconstituted prior to administration. In some cases, the reconstitution is performed using a diluent as described herein.

[0047] In some embodiments, the composition can be administered alone or in combination with a carrier such as a pharmaceutically acceptable carrier or a biocompatible scaffold.

[0048] In some embodiments, the composition is formulated for oral administration. In some embodiments, the composition is an enteric preparation. In some embodiments, the enteric preparation is a dosage form with an enteric coating. For example, the enteric preparation can be an enteric granule, an enteric tablet, or an enteric capsule. In some embodiments, the composition is a capsule. In some embodiments, the capsule is a hard capsule or a soft capsule; or the capsule is a sustained release capsule, a controlled release capsule, or an enteric capsule, or the capsule can be a microencapsulated capsule, or a microcapsule.

[0049] In some embodiments, the composition is any one of a medicine, a health product or a food.

[0050] In some embodiments, the health product or food is any one of a nutritional composition, a food, a candy, a food bar, a food additive, a drink additive, and a dietary supplement.

[0051] In some embodiments, the composition is in a dosage form suitable for infants, children or adults.

[0052] In some embodiments, the composition is in a parenteral dosage form.

[0053] According to a fourth aspect of the present disclosure, there is provided the Lactobacilaceae ( Lactobacillaceae ) strain, the culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillus family strain described in the second aspect or the composition described in the third aspect for use in the preparation of medicines, health products or foods for preventing weight regain or maintaining weight after weight loss.

[0054] In some embodiments, the medicine, health product or food has at least one selected from the following: (1) Maintaining body weight after discontinuation of weight loss medication; (2) prevent blood sugar increases and / or maintain blood sugar balance after discontinuation of weight loss medications; (3) Prevent body fat accumulation after discontinuation of weight loss medication.

[0055] In some embodiments, the body fat includes one or more of subcutaneous fat, epididymal fat, white fat, and visceral fat.

[0056] In some embodiments, the weight loss drug is a GLP-1 receptor agonist.

[0057] In some embodiments, the subject of the application is a subject receiving a GLP-1 receptor agonist for weight loss.

[0058] In some embodiments, the GLP-1 receptor agonist includes one or more of the following: liraglutide or semaglutide.

[0059] The compositions of the present disclosure may also include cellular components, metabolites, secreted molecules and compounds metabolized by the lactobacillus, and the like. Cellular components or cell fractions, metabolites or secreted compounds may be obtained, for example, by recovering the supernatant of a lactobacillus culture or by extracting from a lactobacillus culture; may correspond to components in isolated form from lactobacillus, or any mixture of one or more components from lactobacillus.

[0060] According to the fifth aspect of the present disclosure, provided is the use of the Lactobacillaceae strain described in the first aspect, the culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillaceae strain described in the second aspect, or the composition described in the third aspect in the preparation of a medicine, health product or food for preventing blood sugar increase or maintaining blood sugar balance after stopping weight loss medication.

[0061] According to the sixth aspect of the present disclosure, provided is the use of the Lactobacillaceae strain described in the first aspect, the culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillaceae strain described in the second aspect, or the composition described in the third aspect in the preparation of a medicine, health product or food for preventing, treating, alleviating or alleviating metabolic diseases, metabolic disorders, or diseases caused by metabolic disorders, liver and kidney diseases, cardiovascular and cerebrovascular diseases in a subject in need thereof.

[0062] In some embodiments, the metabolic disease includes at least one of obesity and obesity-related diseases, diabetes, dyslipidemia, glucose intolerance, type I diabetes, type II diabetes, impaired glucose tolerance, insulin resistance, hyperglycemia, hyperinsulinemia, fatty liver, alcoholic fatty liver, hypercholesterolemia, hypertension, hyperlipoproteinemia, hyperlipidemia, hypertriglyceridemia, uremia, ketoacidosis, hypoglycemia, thrombotic diseases, dyslipidemia, non-alcoholic fatty liver disease, and non-alcoholic fatty liver disease.

[0063] As used herein, a microbial population generally refers to a microbial population consisting essentially of a single strain, species or genus, which may be the case when a population is cultured from a subpopulation of such strain, species or genus that is separated and purified. Thus, for a given microbial population, if cultured from an isolated microbial species or strain, such a population will be referred to herein as purified or substantially pure. The resulting population may typically be at least 80% pure for the microbial species or strain, at least 90% pure, at least 95% pure, at least 98% pure, at least 99% pure, at least 99.5% pure, or at least 99.9% pure relative to other microbial species or strains within the particular population. In contrast, the level of non-desirable strains in any particular desired microbial population will be less than 20%, less than 10%, less than 5%, less than 2%, less than 1%, less than 0.5% or less than 0.1%. For example, the level of impurities in a purified population population, such as other undesirable microbial strains or species, may be proportional to or less than the above levels for each desired population. Less than 2%, less than 1%, less than 0.5% or less than 0.1%. In the case where the composition comprises a consortium of multiple microbial populations, each population can have the above-mentioned purities, either before it is incorporated into the composition, or when the consortium is measured in an aggregate. For example, the level of impurities in the purified population consortium, such as other undesirable microbial strains or species, can be proportional to the above-mentioned levels for each desired population or below the above-mentioned levels. Less than 2%, less than 1%, less than 0.5% or less than 0.1%. In the case where the composition comprises a consortium of multiple microbial populations, each population can have the above-mentioned purities, either before it is incorporated into the composition, or when the consortium is measured in an aggregate. For example, the level of impurities in the purified population consortium, such as other undesirable microbial strains or species, can be proportional to the above-mentioned levels for each desired population or below the above-mentioned levels.

[0064] In some embodiments, the causes of obesity and obesity-related diseases include, but are not limited to, at least one of a high-fat diet, a high-sugar diet, high cholesterol, high blood lipids, high blood sugar, NAFLD or NASH.

[0065] In some embodiments, the diabetes includes but is not limited to type I diabetes, type II diabetes, gestational diabetes, HDAC activity-mediated diabetes, diabetic nephropathy, diabetic neuropathy, diabetic eye disease, diabetic retinopathy, diabetic foot, diabetes caused by damage to pancreatic B cells, diabetes caused by insulin resistance, and diabetes caused by obesity.

[0066] In some embodiments, the causes of diabetes include, but are not limited to, at least one of islet cell dysfunction, decreased insulin secretion, increased insulin resistance, a high-fat diet, a high-sugar diet, high cholesterol, hyperlipidemia, hyperglycemia, NAFLD, or NASH.

[0067] In some embodiments, the causes of diabetes include at least one of the following: caused by a high-fat diet, caused by a high-sugar diet, and caused by a high-cholesterol diet.

[0068] In some embodiments, the diabetes mellitus is manifested by hyperglycemia resulting from low levels of insulin and / or peripheral insulin resistance.

[0069] In some embodiments, the metabolic disorder includes but is not limited to: (1) diabetes caused by disordered glucose metabolism, or (2) diabetes caused by abnormal glucose tolerance or decreased glucose tolerance, or (3) diabetes caused by damaged insulin B cells, or (4) diabetes caused by insulin resistance.

[0070] In some embodiments, the liver and kidney diseases include at least one of liver diseases, diseases related to liver damage, and kidney diseases.

[0071] In some embodiments, the cardiovascular and cerebrovascular diseases include atherosclerosis, coronary heart disease, hypertension, cardiovascular diseases in patients with NAFLD or NASH, cardiovascular and cerebrovascular diseases in patients with NAFLD or NASH, and high cholesterol diseases.

[0072] In some embodiments, the liver disease includes at least one of fatty liver, NAFLD, NASH, abnormal liver function, extrahepatic cholestasis, hepatitis, liver injury, intrahepatic cholestasis, liver fibrosis, cirrhosis and liver cell injury.

[0073] In some embodiments, the renal disease comprises at least one of renal injury, abnormal renal function, primary glomerulonephritis, hypertensive arteriolar sclerosis, diabetic nephropathy, secondary glomerulonephritis, tubulointerstitial lesions, and ischemic nephropathy.

[0074] In some embodiments, the cardiovascular disease or cardiovascular and cerebrovascular disease includes, but is not limited to, atherosclerosis, coronary heart disease, hypertension, cardiovascular disease in patients with NAFLD or NASH, cardiovascular and cerebrovascular disease in patients with NAFLD or NASH, and high cholesterol disease.

[0075] In some embodiments, the causes of the cardiovascular disease or cardiovascular and cerebrovascular disease include, but are not limited to, at least one of atherosclerosis, NAFLD, NASH, hyperlipidemia, hyperglycemia or hypercholesterolemia.

[0076] In some embodiments, the drug, health product or food can treat, prevent, alleviate or improve at least one of the following: (1) weight loss, weight control or delayed weight gain; (2) improvement of blood sugar homeostasis imbalance, improvement of blood sugar elevation; (3) prevention, treatment, improvement or alleviation of diabetes; (4) reduction of fat accumulation or reduction of local fat; (5) reduction of blood lipid and / or cholesterol levels; (6) prevention, treatment and improvement of hypertension and atherosclerosis; (7) improvement of intestinal flora; (8) prevention of weight recurrence.

[0077] In some embodiments, the local fat includes one or more of subcutaneous fat, epididymal fat, white fat, visceral fat, inguinal fat, perirenal fat, intermuscular fat, retroperitoneal fat, pericardial fat, and omental fat. BRIEF DESCRIPTION OF THE DRAWINGS

[0078] By reading the detailed description below with reference to the accompanying drawings, the above and other objects, features and advantages of the exemplary embodiments of the present disclosure will become readily understood. In the accompanying drawings, several embodiments of the present disclosure are shown in an exemplary and non-limiting manner, and the same or corresponding reference numerals represent the same or corresponding parts, wherein: Figure 1 The colony morphology of MNH08891 is shown.

[0079] Figure 2 A Gram-stained microscopic morphological photograph of MNH08891 is shown.

[0080] Figure 3 Shown: Electron micrograph of MNH08891.

[0081] Figure 4 The colony morphology of MNH22076 is shown.

[0082] Figure 5 Gram-stained microscopic photographs of strain MNH22076 are shown.

[0083] Figure 6 An electron micrograph of MNH22076 is shown.

[0084] Figure 7 The results of the tolerance of the strains to different pH are shown.

[0085] Figure 8 The results show the tolerance of the strains to different concentrations of NaCl.

[0086] Fig. 9 The results of the tolerance of the strains to different concentrations of bile salts are shown.

[0087] Fig.10 The results of the strains' tolerance to different pH are shown.

[0088] Fig.11 The results show the tolerance of the strains to different concentrations of NaCl.

[0089] Fig.12 The results of the tolerance of the strains to different concentrations of bile salts are shown.

[0090] Fig.13 The phylogenetic tree of strain MNH08891 is shown.

[0091] Fig.14 The phylogenetic tree of strain MNH22076 is shown.

[0092] Fig.15 The figure shows the body weight at the end point of the trial after the cessation of semaglutide treatment. The data are shown as mean ± SD. The statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group.

[0093] Fig.16 The weight gain at the end point of the trial after cessation of semaglutide treatment is shown. The data are presented as mean ± SD. The statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group.

[0094] Fig.17 The weight gain rate after cessation of semaglutide treatment is shown. Data are presented as mean ± SD. Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group.

[0095] Fig.18The blood glucose level at 2 h after cessation of semaglutide treatment is shown. The data are shown as mean ± SD. The statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group, **, p < 0.01 compared with the HFD-Sema group.

[0096] Fig.19 Fasting blood glucose levels after cessation of semaglutide treatment are shown. Data are presented as mean ± SD. Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group, **, p < 0.01 compared with the HFD-Sema group.

[0097] Fig. 20 Subcutaneous fat weight after administration of MNH08891 / MNH22076 after cessation of semaglutide treatment is shown. Data are shown as mean ± SD. Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group, **, p < 0.01 compared with the HFD-Sema group, ***, p < 0.001 compared with the HFD-Sema group.

[0098] Fig.21 Epididymal fat weight after administration of MNH08891 / MNH22076 after cessation of semaglutide treatment is shown. Data are shown as mean ± SD. Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group, **, p < 0.01 compared with the HFD-Sema group, ***, p < 0.001 compared with the HFD-Sema group.

[0099] Fig. 22 White fat of MNH08891 / MNH22076 after cessation of semaglutide treatment is shown. Data are shown as mean ± SD. Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group, **, p < 0.01 compared with the HFD-Sema group, ***, p < 0.001 compared with the HFD-Sema group.

[0100] Fig.23The white fat mass ratio of MNH08891 / MNH22076 after cessation of semaglutide treatment is shown. Data are shown as mean ± SD. Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group, **, p < 0.01 compared with the HFD-Sema group, ***, p < 0.001 compared with the HFD-Sema group.

[0101] Fig.24 Visceral fat after MNH08891 / MNH22076 administration after cessation of semaglutide treatment is shown. Data are shown as mean ± SD. Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group, **, p < 0.01 compared with the HFD-Sema group, ***, p < 0.001 compared with the HFD-Sema group.

[0102] Fig.25 The visceral fat to body weight ratio of MNH08891 / MNH22076 after cessation of semaglutide treatment is shown. Data are shown as mean ± SD. Statistical analysis was performed using the Student's t test; *, p < 0.05 compared with the HFD-Sema group, **, p < 0.01 compared with the HFD-Sema group, ***, p < 0.001 compared with the HFD-Sema group.

[0103] Collection information:

[0104] Lactobacillus paracasei MNH08891 was deposited in Guangdong Microbial Culture Collection Center (GDMCC) on November 28, 2024. The deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, with the deposit number of GDMCC NO.65551. The test result is survival, and the taxonomic name is Lacticaseibacillus paracasei .

[0105] Lactobacillus crispatus MNH22076 was deposited in Guangdong Microbial Culture Collection Center (GDMCC) on November 28, 2024. The deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, with the deposit number GDMCC NO.65554. The test result is survival, and the taxonomic name is Lactobacillus crispatus . DETAILED DESCRIPTION

[0106] The following will be combined with the drawings in the embodiments of the present disclosure to clearly and completely describe the technical solutions in the embodiments of the present disclosure. Obviously, the described embodiments are part of the embodiments of the present disclosure, not all of the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present disclosure.

[0107] The present invention discloses two new strains of Lactobacillus isolated: MNH08891, with a preservation number of GDMCC NO.65551, belonging to Lactobacillus paracasei ( Lacticaseibacillus paracasei ); and MNH22076, deposited with GDMCC NO.65554, belonging to Lactobacillus crispatus of the family Lactobacillaceae ( Lactobacillus crispatus ). Further, the present invention discloses the biochemical properties and therapeutic uses of the strain.

[0108] It is known in the art that bacterial species can be classified and identified by traditional classification methods and molecular biological methods. Traditional classification methods include, but are not limited to, cell morphology observation, Gram staining, flagella staining, various metabolic experiments, etc. Molecular biological methods include, but are not limited to, ribosomal RNA sequence determination methods, determination methods based on whole genome sequencing, etc.

[0109] As used herein, the term "prebiotic" may be a general term referring to chemicals and / or ingredients that can affect the growth and / or activity of microorganisms in a host (e.g., can allow for specific changes in the composition and / or activity of a microbiome).

[0110] The terms "subject," "subject," "individual," "host," and "patient" are used interchangeably herein to refer to any animal subject, including: humans, mammals, laboratory animals, livestock, and domestic pets.

[0111] Compositions or preparations disclosed herein can be used as pharmaceutical preparations, therapeutic compositions, dietary supplements, nutritional supplements, medical probiotics or medical foods. In some cases, the composition is applied in the form of a pharmaceutical preparation. In some cases, the composition is applied in the form of a nutritional supplement. In some cases, the composition is applied in the form of a dietary supplement. In some cases, the composition is applied in the form of a medical food. In some cases, the composition is applied in the form of a medical probiotic. In some cases, the composition (e.g., dietary supplements, nutritional supplements, medical probiotics or medical foods) can be orally administered, for example as a capsule, pill or tablet.

[0112] 16S rRNA is a ribosomal RNA of prokaryotes. The 16S rRNA gene consists of a variable region and a conserved region. The conserved region is common to all bacteria, while the variable region has different degrees of difference between different bacteria. By comparing the 16S rRNA gene sequences of bacteria, according to the base number of sequence differences and their evolutionary distance, an evolutionary tree can be drawn. When the identity between the 16S rRNA gene sequences of two strains is less than 98.65%, they can be judged to belong to different species (see Kim, M., Oh, H.-S., Park, S.-C., & Chun, J. (2014). Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. International Journal of Systematic and Evolutionary Microbiology, 64(Pt 2), 346–351, and Liu, C., Du, M.-X., Abuduaini,R., Yu, H.-Y., Li, D.-H., Wang, Y.-J., Liu, S.-J. (2021). Enlightening the taxonomy darkness of human gut microbiomes with a cultured biobank. Microbiome, 9(1), P23).

[0113] The "identity" between two nucleic acid molecule sequences can be determined using known computer algorithms, such as the "FASTA" program, the GCG program package, BLASTN or FASTA. Commercial or publicly available programs may also be, for example, the DNAStar "MegAlign" program.

[0114] The second generation sequencing technology can also be used to identify bacterial species based on whole genome sequencing, making the identification results more accurate. The average nucleotide identity (ANI) of bacterial genomes refers to the similarity of homologous genes between two bacterial genomes. The ANI value can be calculated by methods such as BLAST. In the field of bacterial taxonomy, it is generally believed that the ANI value needs to reach more than 95% to be identified as belonging to the same species (Jain C, Rodriguez-R LM, Phillippy AM, et al. High throughput ANI analysis of 90K prokaryotic genomes reveals clear species boundaries[J]. Nature Communications, 2018, 9 (1): 5114.).

[0115] Various existing mature ANI value calculation tools can be used, such as local calculation software Jspecies ( / jspecies) and Gegenees ( / documentation.html), online calculation tools ANI caculator (http: / enveomics.gatech.edu / ), EzGenome ( / ezgenome / ani) and ANItools.

[0116] Using the above method, those skilled in the art can determine whether an isolated strain belongs to the lactobacillus species identified by the inventors. For example, when the average nucleotide identity ANI value with MNH08891 is at least 95%, such as 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 97.1%, 97.2%, 97.3%, 97.9%, 97.1 ... When the concentration of the bacterial strain was 7.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%, it can be determined that it belongs to the same strain as MNH08891. species; when the average nucleotide identity ANI value with MNH22076 is at least 95%, for example 95.1%, 95.2%, 95.3%, 95.4%, 95.5%, 95.6%, 95.7%, 95.8%, 95.9%, 96%, 96.1%, 96.2%, 96.3%, 96.4%, 96.5%, 96.6%, 96.7%, 96.8%, 96.9%, 97%, 97.1%, 97.2%, 97.3%, 97.9%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.1%, 99.2%, 99.3%, 99.9%, 99.1%, 99.2%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.1%, 99.2%, 99.3%, 99.9%, 99.1%, 99. .4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%, it can be determined that it belongs to the same species as MNH22076.

[0117] For another example, when its 16S rRNA sequence has at least 98.65% identity with the sequence shown in SEQ ID NO. 1, for example, at least 98.7%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%, it can be determined to belong to the same species as MNH08891; or When its 16S rRNA sequence has at least 98.65% identity with the sequence shown in SEQ ID NO:2, for example, at least 98.7%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%, it can be determined to belong to the same species as MNH22076.

[0118] A "strain" refers to a member of a bacterial species that has a genetic characteristic that allows it to be distinguished from closely related members of the same bacterial species. The genetic characteristic can be the total or partial absence of at least one gene, the total or partial absence of at least one regulatory region (e.g., promoter, terminator, riboswitch, ribosomal binding site), the absence ("cure" of at least one native plasmid), the presence of at least one recombinant gene, the presence of at least one mutated gene, the presence of at least one exogenous gene (a gene from another species), the presence of at least one mutated regulatory region (e.g., promoter, terminator, riboswitch, ribosomal binding site), the presence of at least one non-native plasmid, the presence of at least one antibiotic resistance cassette, or a combination thereof. Genetic characteristics between different strains can be identified by PCR amplification, optionally followed by DNA sequencing of the genomic region of interest or the entire genome. In the case of a strain that gains or loses antibiotic resistance (compared to another strain of the same species) or gains or loses a biosynthetic ability (e.g., an auxotrophic strain), the strains can be distinguished by selection or counter-selection using antibiotics or nutrients / metabolites.

[0119] "Supernatant" or "supernatant" within the meaning herein refers to the culture supernatant of the bacterial strain according to the present disclosure, optionally comprising compounds and / or cell debris of said strain, and / or metabolites and / or molecules secreted by said strain.

[0120] Compositions can be prepared using MNH08891 or MNH22076 described herein, for example, by using pharmaceutically acceptable excipients. The pharmaceutical composition comprises a pharmaceutically effective amount of the lactobacillus, for example, Lactobacillus paracasei with a deposit number of GDMCC NO.65551; or Lactobacillus crispatus of the Lactobacillaceae family with a deposit number of GDMCC NO.65554.

[0121] Suitable pharmaceutically acceptable excipients that may be used include, for example, carriers, excipients, diluents, lubricants, wetting agents, emulsifiers, suspension stabilizers, preservatives, sweeteners and flavors.

[0122] The composition herein can be formulated into any form suitable for enhancing the abundance of lactobacilli in a subject. The composition can be administered orally (e.g., by oral gavage), intramuscular injection, inhalation, intracranial, intralymphatic, intraocular, intraperitoneal, intrapleural, intrathecal, intratracheal, intrauterine, intravascular, intravenous, intravesical, intranasal, intragastrointestinal, bile infusion, cardiac infusion, anterior anus, rectum, spinal subcutaneous, sublingual, topical, intravaginal, transdermal, ureteral or urethral.

[0123] Examples of dosage forms suitable for the composition herein include, but are not limited to, tablets, aerosols, chewable sticks, capsules, capsules containing coated granules, capsules containing sustained-release granules, capsules containing sustained-release granules, and concentrates.

[0124] In some embodiments, the composition is a sugar-coated tablet, a gel capsule, a gel, an emulsion, a tablet, a sheet capsule, a hydrogel, a nanofiber gel, an electrospun fiber, a food bar, a candy, a fermented milk, a fermented cheese, a chewing gum, a powder or a toothpaste, etc.

[0125] In some embodiments, administration can also be by inclusion in a subject's diet, such as inclusion in a functional food for humans or companion animals.

[0126] The compositions provided herein may include a pharmaceutically acceptable excipient, diluent or carrier. The pharmaceutically acceptable excipient, diluent or carrier is well known in the art.

[0127] In some embodiments, the MNH08891 or MNH22076 in the composition of the present disclosure is lyophilized. In some embodiments, the MNH08891 or MNH22076 in the composition of the present disclosure is spray-dried. In some embodiments, the MNH08891 or MNH22076 in the composition of the present disclosure is lyophilized or spray-dried and is alive. In some embodiments, the MNH08891 or MNH22076 in the composition of the present disclosure is lyophilized or spray-dried and can be partially or completely colonized in the intestine. In some embodiments, the lyophilized MNH08891 or MNH22076 is reconstituted before administration. In some embodiments, the reconstitution is carried out using a diluent as described herein.

[0128] In some embodiments, the compositions of the present disclosure are administered orally. Oral administration may involve swallowing, thereby allowing the composition to enter the gastrointestinal tract, and / or administration through the mouth, tongue, or sublingually.

[0129] In some embodiments, the composition is prepared by freeze-drying or spray-drying.

[0130] The compositions disclosed herein include pharmaceutical compositions, health products or foods.

[0131] The subject of the present disclosure may be a human or an animal, and the animal includes but is not limited to cattle, sheep, cats, dogs, horses, rabbits, monkeys, mice, rats, alpacas, camels, etc.

[0132] The pharmaceutical composition disclosed herein can be used to treat, prevent or alleviate metabolic diseases or diseases caused by metabolic disorders.

[0133] In some embodiments, the metabolic disease, metabolic disorder, or disease caused by metabolic disorder includes, but is not limited to, at least one of obesity and obesity-related diseases, cardiovascular disease, diabetes, dyslipidemia, cardiovascular and cerebrovascular disease, glucose intolerance, atherosclerosis, coronary heart disease or hypertension, type I diabetes, type II diabetes, impaired glucose tolerance, insulin resistance, obesity, hyperglycemia, hyperinsulinemia, fatty liver, alcoholic fatty hepatitis, hypercholesterolemia, hypertension, hyperlipoproteinemia, hyperlipidemia, hypertriglyceridemia, hypoglycemia, thrombotic disease, dyslipidemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and atherosclerosis.

[0134] diabetes Diabetes includes type 1 diabetes (T1D), type 2 diabetes (T2D) and gestational diabetes mellitus (GDM). Type 1 diabetes is caused by autoimmune damage or idiopathic reasons. It is characterized by absolute destruction of pancreatic islet function. It mostly occurs in children and adolescents. It must be treated with insulin to achieve satisfactory results, otherwise it will be life-threatening. Type 2 diabetes is a multifactorial syndrome characterized by abnormal carbohydrate / fat metabolism, usually including hyperglycemia, hypertension and abnormal cholesterol. Type 2 diabetes is caused by the inability of insulin to work effectively (low binding to receptors). Therefore, it is necessary not only to check fasting blood sugar, but also to observe blood sugar 2 hours after a meal, especially to do pancreatic islet function tests. There are two types of diabetes during pregnancy. One is that the patient has been diagnosed with diabetes before pregnancy, which is called "diabetes complicated with pregnancy"; the other is that the patient has normal glucose metabolism or potential glucose tolerance before pregnancy, and the diabetes that appears or is diagnosed during pregnancy, which is also called "gestational diabetes (GDM)". More than 80% of pregnant women with diabetes have GDM.

[0135] Cardiovascular disease TG (triglycerides) are mainly involved in energy metabolism in the human body and generate heat energy. Excessive TG content in the blood can lead to blood viscosity, causing lipids to deposit on the blood vessel walls, gradually forming small plaques, namely atherosclerosis. Increased LDL-C is the main and independent risk factor for the occurrence and development of atherosclerosis; the increased level of LDL-C is also an indicator for measuring coronary heart disease. Since HDL-C can transport cholesterol in the blood vessel wall to the liver for decomposition and metabolism (i.e., reverse cholesterol transport), it can reduce the deposition of cholesterol in the blood vessel wall and play an anti-atherosclerotic role.

[0136] obesity Obesity refers to a certain degree of obvious overweight and excessive fat layer. It is a state caused by excessive accumulation of body fat, especially triglycerides. It is an abnormal or excessive accumulation of fat that poses a risk to health. Excessive accumulation of body fat due to excessive food intake or changes in body metabolism causes excessive weight gain and causes pathological, physiological changes or lurking in the human body. A body mass index of more than 25 is considered overweight, and more than 30 is considered obese. Obesity increases the risk of many physical and mental diseases. It is mainly related to metabolic syndrome, a combination of diseases including type 2 diabetes, hypertension, hypercholesterolemia and hypertriglyceridemia. In general, the effects of obesity on health can be divided into two categories: diseases that can be attributed to increased body fat (such as osteoarthritis, obstructive sleep apnea, etc.) and diseases that increase the number of fat cells (diabetes, dyslipidemia, cancer, cardiovascular disease, non-alcoholic fatty liver or non-alcoholic steatohepatitis, etc.). The "obesity-related diseases" can be selected from the following diseases: overeating, binge eating, bulimia, hypertension, diabetes, elevated plasma insulin concentration, insulin resistance, hyperlipidemia, metabolic syndrome, insulin resistance syndrome, obesity-related gastroesophageal reflux disease, atherosclerosis, hypercholesterolemia, hyperuricemia, low back pain, cardiac hypertrophy and left ventricular hypertrophy, lipodystrophy, non-alcoholic steatohepatitis, cardiovascular disease, and polycystic ovary syndrome, as well as subjects with these obesity-related diseases including those who wish to lose weight.

[0137] The obesity-related diseases in the present disclosure include at least one of the following diseases: obesity, metabolic syndrome, cardiovascular disease, hyperlipidemia, hypercholesterolemia, hypertension, insulin resistance syndrome, obesity-related gastroesophageal reflux disease and steatohepatitis.

[0138] The pharmaceutical composition disclosed herein can be used to treat, prevent or improve metabolic disorders after discontinuation of GLP-1 receptor agonists.

[0139] In some embodiments, the metabolic disease includes metabolic diseases, metabolic disorders, or diseases caused by metabolic disorders, including but not limited to: obesity and obesity-related diseases, cardiovascular diseases, cardiovascular and cerebrovascular diseases, diabetes, dyslipidemia, glucose intolerance, type I diabetes, type II diabetes, impaired glucose tolerance, insulin resistance, obesity, hyperglycemia, hyperinsulinemia, fatty liver, alcoholic fatty hepatitis, hypercholesterolemia, hypertension, hyperlipoproteinemia, hyperlipidemia, hypertriglyceridemia, hypoglycemia, thrombotic diseases, dyslipidemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), at least one of atherosclerosis.

[0140] In some embodiments, the metabolic disorder comprises weight gain, imbalanced blood glucose homeostasis, and fat accumulation.

[0141] Disturbance of blood sugar homeostasis These include abnormal blood sugar metabolism, insulin resistance, abnormal postprandial blood sugar levels, or abnormal fasting blood sugar levels.

[0142] Fat accumulation Fat accumulation includes subcutaneous fat, epididymal fat, white fat, visceral fat, inguinal fat, perirenal fat, intermuscular fat, retroperitoneal fat, pericardial fat, and omental fat accumulation.

[0143] GLP-1 receptor agonists are compounds that have agonist activity on the glucagon-like peptide-1 receptor. Their effects of increasing insulin secretion by binding to the GLP-1 receptor were revealed.

[0144] In some embodiments, the GLP-1 receptor agonist includes one or more of the following: Exenatide, Liraglutide, Lixisenatide, Dulaglutide, Albiglutide, Semaglutide, Betaglutide, Losenapeptide, and Somalutide.

[0145] The present disclosure provides a Lactobacillaceae strain, a culture thereof, a live bacterium, a freeze-dried bacterium or an inactivated bacterium or a composition thereof for use in preparing a medicine, a health product or a food for preventing weight regain or maintaining weight after weight loss.

[0146] In some embodiments, the medicine, health product or food has at least one selected from the following: (1) Maintaining body weight after discontinuation of weight loss medication; (2) prevent blood sugar increases and / or maintain blood sugar balance after discontinuation of weight loss medications; (3) Prevent body fat accumulation after discontinuation of weight loss medication.

[0147] In some embodiments, the body fat includes one or more of subcutaneous fat, epididymal fat, white fat, and visceral fat.

[0148] In some embodiments, the weight loss drug is a GLP-1 receptor agonist.

[0149] In some embodiments, the subject of the application is a subject receiving a GLP-1 receptor agonist for weight loss.

[0150] In some embodiments, the GLP-1 receptor agonist includes one or more of the following: liraglutide or semaglutide.

[0151] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature of the art or in accordance with the product or instrument instructions are used. All reagents or instruments are indicated by the manufacturer and can be purchased commercially.

[0152] Example

[0153] The following are non-limiting examples for practicing the present disclosure. The following examples are provided merely to illustrate embodiments of the present disclosure and should not be construed as limiting the present disclosure in any way.

[0154] The culture medium involved in the embodiment includes: Anaerobic blood agar plate: purchased from Huankai Microorganisms, formula: casein pancreatic digest 10g / L, heart pancreatic digest 3g / L, corn starch 1g / L, meat gastric digest 5g / L, yeast extract powder 5g / L, sodium chloride 5g / L, agar 15g / L, sterile defibrinated sheep blood 50~100mL / L, pH7.3±0.2.

[0155] MRS medium: peptone 10.0 g, beef extract powder 10.0 g, yeast extract powder 5.0 g, glucose 20.0 g, Tween-80 1.0 mL, dipotassium hydrogen phosphate 2.0 g, sodium acetate 5.0 g, triammonium citrate 2.0 g, magnesium sulfate (MgSO4·7H2O) 0.1 g, manganese sulfate (MnSO4·4H2O) 0.05 g.

[0156] The above culture medium can be prepared by conventional preparation methods and sterilization methods.

[0157] Example 1. Isolation and identification of strains

[0158] 1.1 Isolation of strains MNH08891 and MNH22076 The two intestinal lactobacilli strains, Lactobacillus paracasei MNH08891 and Lactobacillus crispatus MNH22076, screened by the present invention are both isolated from fecal samples of healthy volunteers.

[0159] Specifically, the strain isolation method is as follows: The donor takes 2 to 5 g of fresh feces and puts it into a sample collection and storage tube. After shaking and homogenization, the processed feces sample is placed in an ice box and sent to the laboratory for strain isolation within 24 hours.

[0160] Dispense physiological saline in a biosafety cabinet, 9 mL / tube; prepare anaerobic blood agar plates for strain isolation culture medium, and transfer them to the anaerobic workstation 24 hours in advance, marking the sample information, culture medium type, isolation date, etc.

[0161] Take a fresh stool sample and place it in an anaerobic workstation and shake it with a vortex shaker for 1 min to mix it. Pipette 1 mL of the sample into 9 mL of normal saline and mix it for 10 min. -1 dilution, and then serially diluted to 10 -6 Dilution solution for later use.

[0162] Take 10 -6 Drop the diluted solution onto the anaerobic blood agar plate at a rate of 100 μL / plate. Spread evenly. After the surface of the plate is dry, invert the plate and culture at 37°C for 3 to 5 days.

[0163] 1.2 Purification of strains Observe the growth of the strain and pick a single colony with a sterilized toothpick to purify the strain. The purified strain is placed at 37°C for anaerobically culture.

[0164] The pure culture strain was prepared into 20% glycerol / water-bacteria solution and stored at -80℃.

[0165] 1.3 Strain preservation Lactobacillus paracasei MNH08891 was deposited in the Guangdong Microbial Culture Collection Center (GDMCC) on November 28, 2024. The storage address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, the storage number is GDMCC NO.65551, the test result is survival, and the taxonomic name is Lacticaseibacillusparacasei.

[0166] Lactobacillus crispatus MNH22076 was deposited in the Guangdong Microbial Culture Collection Center (GDMCC) on November 28, 2024. The storage address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, the storage number is GDMCC NO.65554, the test result is survival, and the taxonomic name is Lactobacillus crispatus.

[0167] 1.4 Morphological characteristics of strains Morphological characteristics of strain MNH08891 The strain MNH08891 was inoculated into MRS medium and cultured anaerobically at 37°C for 48 h. Visible colonies were formed on the MRS plate medium. The colonies were round, with regular and smooth edges, about 1-3 mm in diameter, white, and opaque. The strain was Gram-positive. Microscopic morphological observation revealed that it had no flagella, was non-motile, rod-shaped, and about 0.5-1µm×1-2µm in size. The colony morphology of strain MNH08891 cultured on MRS plate for 48 h is shown in Figure 1 Gram staining microscopic morphology of strain MNH08891 is shown in Figure 2 , Gram staining positive. Electron microscopic photos are shown in Figure 3 .

[0168] Morphological characteristics of strain MNH 22076 The strain MNH22076 was inoculated into MRS medium and cultured anaerobically at 37°C for 48 h. Visible colonies were formed on the MRS plate medium. The colonies were round, with regular and smooth edges, about 1-2 mm in diameter, white, and opaque. The strain was Gram-positive. Microscopic morphological observation revealed that it had no flagella, was non-motile, rod-shaped, and about 0.5-1 µm × 1.5-4 µm in size. The colony morphology of strain MNH 22076 cultured on MRS plate for 48 h is shown in Figure 4 Gram staining microscopic morphology of strain MNH22076 is shown in Figure 5 , Gram staining positive. Electron microscopic photos are shown in Figure 6 .

[0169] 1.5. Physiological and biochemical characteristics of strains The strain MNH08891 can grow in the pH range of 4.0 to 10.0, with the optimal growth pH being 6.0 to 9.0 (see the results of strain tolerance to different pH values). Figure 7 ); it does not grow on medium containing more than 5% w / w NaCl (the results of strain tolerance to different concentrations of NaCl are shown in Figure 8 ); strain MNH08891 can survive and grow in the bile salt concentration range of 0% to 0.25%, and cannot grow when the bile salt concentration is greater than or equal to 0.3% (the strain's tolerance to different concentrations of bile salts is shown in Fig. 9 ). This indicates that strain MNH08891 is tolerant to salt concentrations less than 5% and choline concentrations less than 0.3%.

[0170] Physiological characteristics of strain MNH22076 The strain MNH22076 can grow in the range of pH 5.0 to 10.0, with the optimal growth pH being 8.0 (see the results of strain tolerance to different pH values). Fig.10 ); it can still grow in a medium with a NaCl content of more than 6% (the results of the strain's tolerance to different concentrations of NaCl are shown in Fig.11 ); strain MNH22076 can survive and grow in the bile salt concentration range of 0% to 0.1%, and cannot grow when the bile salt concentration is greater than or equal to 0.15% (the strain's tolerance to different concentrations of bile salts is shown in Fig.12 ). This indicates that strain MNH22076 is tolerant to salt concentrations less than 6% and choline concentrations less than 0.15%.

[0171] 1.6. Results of biochemical identification of strain MNH08891 using API 50CHL The strain MNH08891 was biochemically identified using the Lactobacillus kit API 50CHL (Mérieux, CN5041010) according to the instructions to detect the acid-producing carbon source. The culture conditions were: 37°C, anaerobic. The experimental results are shown in Table 1.

[0172] As shown in Table 1, strain MNH08891 can utilize ribose, galactose, glucose, fructose, mannose, dulose, mannitol, sorbitol, methyl-D-glucose, N-acetylglucosamine, starch, arabinitol, esculin, salicin, cellobiose, maltose, lactose, sucrose, trehalose, melezitose, gentiobiose, turanose, and tagatose to ferment and produce acid, so these substances and their derivatives can be used as carbon sources during the fermentation or cultivation of strain MNH08891.

[0173]

[0174] As shown in Table 1, strain MNH08891 can utilize ribose, galactose, glucose, fructose, mannose, dulose, mannitol, sorbitol, methyl-D-glucose, N-acetylglucosamine, starch, arabinitol, esculin, salicin, cellobiose, maltose, lactose, sucrose, trehalose, melezitose, gentiobiose, turanose, and tagatose to ferment and produce acid, so these substances and their derivatives can be used as carbon sources during the fermentation or cultivation of strain MNH08891.

[0175] 1.7 Results of biochemical identification of strain MNH 22076 using API 50CHL.

[0176] The experimental results of strain MNH 22076 are shown in Table 2.

[0177] As shown in Table 2, strain MNH22076 can utilize galactose, glucose, fructose, mannose, methyl-D-glucose, N-acetylglucosamine, starch, arabinitol, esculin, salicin, cellobiose, maltose, lactose melibiose, sucrose, trehalose, inulin, melezitose, raffinose, starch, and gentiobiose to ferment and produce acid, so these substances and their derivatives can be used as carbon sources during the fermentation or cultivation of strain MNH22076.

[0178]

[0179] As shown in Table 2, strain MNH22076 can utilize galactose, glucose, fructose, mannose, methyl-D-glucose, N-acetylglucosamine, starch, arabinitol, esculin, salicin, cellobiose, maltose, lactose melibiose, sucrose, trehalose, inulin, melezitose, raffinose, starch, and gentiobiose to ferment and produce acid, so these substances and their derivatives can be used as carbon sources during the fermentation or cultivation of strain MNH22076.

[0180] 1.7. Minimum inhibitory concentration test of bacterial strain antibiotics

[0181] Minimum inhibitory concentration test of antibiotics for strain MNH08891 The E-test (Liofilchem) paper strip was used to determine the antibiotic minimum inhibitory concentration of MNH08891 according to the instructions. The test results are shown in Table 3.

[0182]

[0183] The results are shown in Table 3. The strain MNH08891 is sensitive to most types of antibiotics, and the risk of subjects developing antibiotic resistance due to long-term use of MNH19250 is low.

[0184] 1.8 Antibiotic Minimum Inhibitory Concentration Test for Strain MNH22076 The minimum inhibitory concentration of antibiotics for MNH22076 was determined using E-test (Liofilchem) paper strips. The test results are shown in Table 4.

[0185]

[0186] The results are shown in Table 4. The strain MNH22076 is sensitive to most types of antibiotics, and the risk of subjects developing antibiotic resistance due to long-term use of MNH22076 is low.

[0187] 1.9 Identification of strains 1.9.1. 16S rRNA gene amplification Fresh cultures of strains MNH08891 and MNH22076 were taken to extract genomic DNA of the strains, and the extracted genomic DNA of the strains was used as a template for 16S rRNA gene amplification.

[0188] The primer pairs used in the 16S rRNA gene PCR of the present invention are: 27F: 5'-AGAGTTTGATCMTGGCTCAG-3' (SEQ ID No: 3) 1492R: 5'-TACGGYTACCTTGTTACGACTT-3' (SEQ ID No: 4).

[0189] The PCR reaction procedure is as follows: Pre-denaturation: 94℃, 4 min; Denaturation: 94℃, 50 sec; Annealing: 52℃, 40 sec; Extension: 72℃, 70 sec; Final extension: 72℃, 10 min. (36 cycles).

[0190] 1.9.2. 16S rRNA gene sequencing After PCR amplification, the PCR product was purified and sent to Sangon Biotechnology Co., Ltd. for 16S rRNA gene sequencing to obtain the 16S rRNA gene sequence.

[0191] The 16S rRNA gene sequence of strain MNH08891 is shown in SEQ ID No: 1:

[0192] The 16S rRNA gene sequence of strain MNH 22076 is shown in SEQ ID No: 2:

[0193] 1.9.3. Strain identification results The strain 16S rRNA gene sequence returned by sequencing was submitted to the NCBI Basic Local AlignmentSearch Tool for strain 16S rRNA gene analysis to confirm the strain classification information.

[0194] The measured sequences were analyzed with the data in GenBank by BLAST. The comparison results showed that the strain with the highest similarity to MNH08891 was Lacticaseibacillus paracasei, with a similarity of 100%. Whole genome ANI analysis showed that the similarity with Lacticaseibacillus paracasei (GCF_000829035.1) was 98.14%, and the alignment score (AF) was 89%. Therefore, strain MNH08891 was a strain of Lacticaseibacillus paracasei. The strain with the highest similarity to MNH22076 was Lactobacillus crispatus, with a similarity of 100%. Whole genome ANI analysis showed that the similarity with Lactobacillus crispatus (GCF 002088015.1) was 97.63%, and the alignment score (AF) was 86%. Therefore, strain MNH22076 was a strain of Lactobacillus crispatus.

[0195] The 16S rRNA gene sequences of MNH08891 and MNH22076 were compared with those of Lacticaseibacillus related strains retrieved from GenBank and other databases, and phylogenetic trees were constructed respectively.

[0196] The sequences of MNH08891 and MNH22076 and the model strains with high 16S rRNA gene sequence similarity in the NCBI database were aligned, and then the phylogenetic tree was constructed using the software MEGA 5 (the phylogenetic tree was constructed using the maximum likelihood method). The phylogenetic tree nodes in the figure only show values ​​with Bootstrap values ​​greater than 50%.

[0197] From the phylogenetic tree ( Fig.13 ) It can be seen that strain MNH08891 is clustered with Lacticaseibacillus paracasei and is a new strain of Lacticaseibacillus paracasei.

[0198] From the phylogenetic tree ( Fig.14) It can be seen that strain MNH22076 is clustered with Lactobacillus crispatus and is a new strain of Lactobacillus crispatus.

[0199] 1.10 Strain genome analysis The genome of the original strain MNH08891 was fragmented by ultrasound, with a fragment length range of ~350bp, and then an Illumina sequencing library was constructed using a standard DNA library construction kit (NEB UltraTM). The constructed sequencing library was sequenced with NovaSeq (Illumina) for double-end 150bp. 1.12Gbp of data was obtained by sequencing, of which Q20 accounted for 96.19%.

[0200] The raw sequencing data of the genome were filtered using fastp (version: 0.20.0) with filtering parameters: "--poly_g_min_len 10 --poly_x_min_len 10 -q 15 -u 40 -n 5 -l 50". The filtered raw data were assembled using SPAdes (version: v3.14.0) with assembly parameters "--isolate --cov-cutoff 10". The total gene length of the genome assembly was 3.02 Mbp, the N50 length was 98.7 kbp, and the GC content was 46.26%.

[0201] The genomic genes were predicted and analyzed using the prokaryotic analysis software genome annotation pipeline prokka (version: 1.14.5) with the parameters "--gcode 11 --evalue 1e-09". A total of 2911 CDS sequences were predicted, with an average CDS sequence length of 881 bp.

[0202] Potential antibiotic resistance genes in the genome were analyzed using the RGI pipeline (version: 4.2.2), and the antibiotic resistance gene database was CARD (version: 3.0.0, https: / / card.mcmaster.ca / analyze / rgi). No resistance genes were found in the comparison information. The analysis of potential virulence factors and related genes in the genome was performed using NCBI blastp (version: 2.7.1+) to compare with the virulence factor database VFDB (virulence factor database, http: / / www.mgc.ac.cn / cgi-bin / VFs / v5 / main.cgi, updated on September 19, 2019). The analysis of potential secondary metabolism gene clusters in the genome was performed using antiSMASH6 (version: 6.0.1).

[0203] The analysis of potential primary metabolic gene clusters in the genome was performed using gutSMASH5 (version: 1.0.0). The detailed alignment results are shown in Table 5.

[0204]

[0205] The analysis of potential primary metabolic gene clusters in the genome was performed using gutSMASH5 (version: 1.0.0). The detailed alignment results are shown in Table 6.

[0206]

[0207] The results in Tables 5 and 6 show that a gene cluster capable of producing primary metabolites that convert pyruvate into acetate and formate was identified in strain MNH08891.

[0208] 1.11 Genome analysis of strain MNH 22076 The genome of the original strain MNH22076 was fragmented by ultrasonic method, with the fragment length ranging from 350 bp, and then the Illumina sequencing library was constructed using a standard DNA library construction kit (NEB UltraTM). The constructed sequencing library was sequenced with NovaSeq (Illumina) for double-end 150 bp. The sequencing obtained 1.28 Gbp of data, of which Q20 accounted for 95.28%.

[0209] The raw sequencing data of the genome were filtered using fastp (version: 0.20.0) with filtering parameters: "--poly_g_min_len 10 --poly_x_min_len 10 -q 15 -u 40 -n 5 -l 50". The filtered raw data were assembled using SPAdes (version: v3.14.0) with assembly parameters "--isolate --cov-cutoff 10". The total gene length of the genome assembly was 2.28 Mbp, the N50 length was 33.0 kbp, and the GC content was 36.59%.

[0210] The genomic genes were predicted and analyzed using the prokaryotic analysis software genome annotation pipeline prokka (version: 1.14.5) with the parameters "--gcode 11 --evalue 1e-09". A total of 2257 CDS sequences were predicted, with an average CDS sequence length of 890 bp.

[0211] Potential antibiotic resistance genes in the genome were analyzed using the RGI pipeline (version: 4.2.2), where the antibiotic resistance gene database was CARD (version: 3.0.0, https: / / card.mcmaster.ca / analyze / rgi). No resistance genes were found after comparison) The analysis of potential virulence factors and related genes in the genome was performed using NCBI blastp (version: 2.7.1+) to compare the virulence factor database VFDB (virulence factor database, http: / / www.mgc.ac.cn / cgi-bin / VFs / v5 / main.cgi, updated on September 19, 2019). The detailed comparison results are shown in Table 7.

[0212]

[0213] The analysis of potential primary metabolic gene clusters in the genome was performed using gutSMASH5 (version: 1.0.0). The detailed alignment results are shown in Table 8.

[0214]

[0215] The results in Tables 7 and 8 show that a gene cluster capable of producing a primary metabolite that converts arginine into bicarbonate was identified in strain MNH22076.

[0216] Example 2. Metabolic Experiment

[0217] This example conducted an animal experiment on the effect of MNH08891 and MNH22076 on preventing obesity relapse in an obese mouse model after cessation of semaglutide treatment.

[0218] Mice were fed a high-fat diet for 10 weeks, and then induced to be obese and treated with semaglutide for 4 weeks before discontinuation of the drug. MNH08891 and MNH22076 were used for intervention after discontinuation of semaglutide treatment to investigate the effect of MNH08891 and MNH22076 on preventing obesity after discontinuation of semaglutide treatment. This invention has passed the ethical review of the Animal Ethics Committee of Muen Biological.

[0219] The feed involved in this embodiment includes: The basic feed product number is D12451B, which was purchased from Parker Biotechnology Co., Ltd.

[0220] The high-fat feed was purchased from Parker Biotechnology Co., Ltd. with the product number D12492.

[0221] 1. Experimental Methods 1) Experimental animals: C57BL / 6J mice were purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0222] 2) Test strains: After thawing the glycerol cryopreserved tubes of MNH08891 and MNH22076 strains at 37°C, inoculate them into MRS medium in a biosafety cabinet for activation. The activated strains were inoculated into MRS liquid medium for cultivation to obtain a sufficient number of cultures. The cultured bacterial solution was concentrated by centrifugation and resuspended in PBS containing 25% glycerol and 0.05% L-Cys HCl to obtain the purity and viable count (2×10 9 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0223] 3) Negative control: Use PBS containing 25% glycerol and 0.05% L-Cys HCl as the negative control.

[0224] 4) Experimental process: After the quarantine period, 8 C57BL / 6J male mice aged 5-6 weeks were randomly selected to be fed with a basic diet, and the remaining mice were all fed with a high-fat diet for 10 weeks. After 10 weeks, all mice fed with a basic diet were transferred to the NCD group. 32 mice with a body weight range of 36g-44g were selected from the mice fed with a high-fat diet, and randomly stratified and grouped according to body weight, 8 mice / group. A total of 4 groups were divided into HFD group, HFD+Sema group, MNH08891 group and MNH22076 group. The NCD group continued to be fed with a basic diet, and the other 4 groups continued to be fed with a high-fat diet. Drug administration began after grouping (D1), the NCD group and the HFD group were given a negative control, and the remaining three groups were given Semaglutide (30nmol / kg, once a day), and Semaglutide was stopped after 4 weeks of administration. The HFD+Sema group began to receive the negative control, the MNH08891 group and the MNH22076 group began to be gavaged with MNH08891 and MNH22076, and the NCD group and the HFD group continued to receive the negative control, which was gavaged once a day, 200 μl each time, for a total of 28 days. During the experiment, the mice had free access to water and food, and a 12h / 12h day and night cycle was adopted. During the experiment, a general clinical observation was performed once after each administration. The endpoint of this experiment was the day after the end of administration (D57). During the experiment, a general clinical observation was performed once after each administration.

[0225] Experiments on fasting blood glucose and oral glucose tolerance in mice: Oral glucose tolerance test (OGTT): In the last week of administration, measure the OGTT after fasting for 12 hours (e.g. fasting from 20:30:00 in the evening to 08:30:00 the next day). Weigh the fasting weight of mice, and gavage glucose according to the fasting weight of mice. The gavage glucose dose is 2g / kg (g of glucose / kg of fasting weight of mice). Measure fasting blood glucose and blood glucose values ​​15min, 30min, 60min, 90min, and 120min after sugar administration. Strictly time each mouse and accurately measure blood glucose values ​​at 6 time points.

[0226] Subcutaneous fat, epididymal fat, perirenal fat, and mesenteric fat were directly dissected and weighed for measurement; visceral fat was the sum of epididymal fat, perirenal fat, and mesenteric fat; and white fat was the sum of all fat.

[0227] At the end of the trial, the samples were collected according to the protocol, and the data were summarized and analyzed for each anatomical data and serum test data. All data were expressed in the form of Mean±SD, and GraphPad Prism software was used for drawing and statistical analysis. For pairwise comparisons, the t test (Student's t test) analysis method was used. No significance is indicated; significant differences are indicated by *, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0228] 1.1 MNH08891 and MNH22076 can improve weight regain after cessation of semaglutide treatment Figure 15-17 The results showed that MNH08891 and MNH22076 could reduce weight gain and weight gain rate after discontinuation of semaglutide treatment, indicating that MNH08891 and MNH22076 could improve weight regain after discontinuation of semaglutide treatment and have the purpose of preventing weight regain.

[0229] 1.2 MNH08891 / MNH22076 can improve the increase in blood sugar after cessation of semaglutide treatment Figure 18-19 The results showed that MNH08891 and MNH22076 could reduce AUC-OGTT after discontinuation of semaglutide treatment, significantly reduce blood glucose 2 hours after meal, and significantly reduce fasting blood glucose at the test endpoint, indicating that MNH08891 and MNH22076 could improve the imbalance of blood glucose homeostasis after discontinuation of semaglutide treatment, improve blood glucose increase, and have the use of preventing blood glucose increase and maintaining blood glucose balance.

[0230] 1.3 MNH08891 / MNH22076 can improve fat accumulation after cessation of semaglutide treatment Figures 20 to 25The results showed that MNH08891 and MNH22076 could significantly reduce the weight of subcutaneous fat, epididymal fat, white fat, and visceral fat after discontinuation of semaglutide treatment, and significantly reduce the proportion of white fat and visceral fat, indicating that MNH08891 and MNH22076 could improve fat accumulation after discontinuation of semaglutide treatment, and could prevent body fat accumulation and weight regain.

[0231] In summary, the above examples illustrate that the new lactobacilli MNH08891 and MNH22076 isolated from fecal samples of healthy volunteers are Gram-positive anaerobic bacteria, tolerant to salt and choline, and easy to colonize. Lactobacilli MNH08891 and MNH22076 can produce acid using a variety of carbon sources and are easy to culture; they are sensitive to most types of antibiotics and have a low risk of antibiotic resistance.

[0232] Lactobacillus MNH08891 and MNH22076 can reduce weight gain and weight gain rate after discontinuation of semaglutide treatment; improve blood sugar homeostasis imbalance after discontinuation of semaglutide treatment, and improve blood sugar increase; significantly reduce fat accumulation after discontinuation of semaglutide treatment, and have the uses of preventing blood sugar increase and fat accumulation in the body, preventing weight regain, and maintaining blood sugar balance.

[0233] Although multiple embodiments of the present disclosure have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Those skilled in the art may think of many changes, modifications, and alternatives without departing from the thought and spirit of the present disclosure. It should be understood that in the process of practicing the present disclosure, various alternatives to the embodiments of the present disclosure described herein may be adopted. The attached claims are intended to define the scope of protection of the present disclosure, and therefore cover equivalents or alternatives within the scope of these claims.

Claims

1. A lactobacillus family that prevents obesity ( Lactobacillaceae ) strain, characterized in that It is deposited in Guangdong Provincial Microbiological Culture Collection Center with a deposit number of GDMCC No: 65551 or a deposit number of GDMCC No: 65554.

2. The culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillusaceae strain according to claim 1.

3. The culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillusaceae strain according to claim 2, characterized in that: The culture includes the fermentation broth of the Lactobacillus strain or a concentrated or dried product of the fermentation broth.

4. A composition for preventing obesity, characterized in that: It comprises an effective amount of the culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillus strain according to claim 1 or any one of claims 2-3; wherein the culture comprises any one of the following A) to D): A) fermentation broth of the strain; B) the supernatant of the fermentation broth of the strain; C) an inactivated fermentation broth of the strain; D) The concentrated or dried product of any one of A) to C) above.

5. The composition according to claim 4, characterized in that The composition further comprises one or more pharmaceutically acceptable carriers, food carriers, active agents, excipients and / or auxiliary materials.

6. The composition according to claim 5, characterized in that The active agent is one or more of probiotics and prebiotics.

7. The composition according to claim 4, characterized in that The composition is any one of medicine, health care product or food.

8. The composition according to claim 7, characterized in that The health care product or food is any one of a nutritional composition, a food, a candy, a food bar, a food additive, a drink additive, and a dietary supplement.

9. The Lactobacillus family as claimed in claim 1 ( Lactobacillaceae ) strain, the culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillusaceae strain according to any one of claims 2 to 3, or the composition according to any one of claims 4 to 8 for use in the preparation of a medicine, health product or food for preventing weight regain or maintaining weight after weight loss.

10. The use according to claim 9, characterized in that The medicine, health product or food is used for at least one selected from the following: (1) Maintaining body weight after discontinuation of weight loss medication; (2) prevent blood sugar increases and / or maintain blood sugar balance after discontinuation of weight loss medications; (3) Prevent body fat accumulation after discontinuation of weight loss medication.

11. The use according to claim 10, characterized in that The body fat includes one or more of subcutaneous fat, epididymal fat, white fat, and visceral fat.

12. The use according to claim 10 or 11, characterized in that: The subject of the application is a subject who receives a GLP-1 receptor agonist for weight loss.

13. The use according to claim 12, characterized in that: The GLP-1 receptor agonist includes one or more of the following: liraglutide or semaglutide.

14. The Lactobacillus family as claimed in claim 1 ( Lactobacillaceae ) strain, the culture, live bacteria, freeze-dried bacteria or inactivated bacteria of the Lactobacillus family strain according to any one of claims 2-3, or the composition according to any one of claims 4-8 in the preparation of a medicine, health product or food for preventing blood sugar from rising or maintaining blood sugar balance after stopping weight loss medication.

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