Compound lactic acid bacteria composition and application thereof

Abstract

The invention discloses a compound lactic acid bacteria composition and application thereof, and belongs to the technical field of microbial medicine. The compound lactic acid bacteria composition is prepared from lactobacillus plantarum (Lactobacillus plantarum) HMPM1006C and lactobacillus curvatus (Lactobacillus curvatus) HMPM1403C, and the ratio of the viable count of the lactobacillus plantarum (Lactobacillus plantarum) HMPM1006C to the viable count of the lactobacillus curvatus (Lactobacillus curvatus) HMPM1403C to the viable count of the lactobacillus plantarum (Lactobacillus plantarum) HMPM1403C is 1 to (0.1 to 3). Through the synergistic effect of multiple target points, absorption of carbohydrates in the intestinal tract can be delayed, insulin sensitivity can be improved, cholesterol metabolism can be promoted, meanwhile, intestinal flora balance is adjusted, inflammatory response is relieved, improvement on type 2 diabetes mellitus and related metabolic diseases is achieved, and the effect is remarkably superior to that of a single strain; a safe and effective new strategy is provided for prevention and adjuvant treatment of metabolic syndrome, type 2 diabetes and obesity.

Description

Technical Field

[0001] This application relates to the field of microbial pharmaceutical technology, specifically to a compound lactic acid bacteria composition and its application. Background Technology

[0002] Type 2 diabetes is a complex metabolic disease characterized by insulin resistance and glucose metabolism disorders, often accompanied by lipid metabolism abnormalities, and has become a major global public health problem. Current mainstream drug treatments, such as oral hypoglycemic agents and insulin, while controlling blood sugar, have limitations including gastrointestinal side effects, the risk of hypoglycemia, weight gain, and drug dependence, and cannot fundamentally reverse metabolic imbalances. Therefore, developing safe, effective, and naturally derived novel adjunctive intervention strategies has urgent market demand and clinical value.

[0003] The gut microbiota plays a central role in regulating host glucose and lipid metabolism. Type 2 diabetes patients commonly exhibit gut microbiota dysbiosis, characterized by reduced abundance and diversity of beneficial bacteria. This leads to insufficient production of short-chain fatty acids, impaired intestinal barrier function, and chronic inflammation, exacerbating metabolic disorders. Therefore, probiotics, as active microorganisms that regulate the balance of the gut microbiota, demonstrate significant potential in improving metabolic health.

[0004] In the prior art, various probiotic strains with single hypoglycemic or lipid-lowering activities have been reported. For example, CN114621896A discloses a strain of *Lactobacillus plantarum* 84-3, which achieves hypoglycemic and lipid-lowering effects by reducing fasting blood glucose in diabetic rats and regulating inflammatory factors and short-chain fatty acid metabolism; CN114958651A discloses the extracellular polysaccharide of *Lactobacillus plantarum* DPA1C, which has in vitro inhibitory activity against α-glucosidase and α-amylase; and CN120137823A discloses *Lactobacillus fermentum* YZU-lbfel, whose co-fermented product with kombucha can improve impaired glucose tolerance and insulin resistance. In addition, some single strains, such as *Lactobacillus plantarum* and *Bifidobacterium longum*, have also been shown to have certain metabolic regulatory activities, but the target and efficacy of single strains are limited, making it difficult to comprehensively and stably correct the complex multiple pathological processes of type 2 diabetes (such as simultaneously improving blood glucose, blood lipids, inflammation, and intestinal barrier function).

[0005] Existing technologies lack probiotic formulations designed to synergistically target the multiple pathological mechanisms of type 2 diabetes. Single strains or single active ingredients cannot achieve synergistic intervention across multiple targets and mechanisms, thus failing to provide novel probiotic compositions that can more efficiently and comprehensively improve glucose and lipid metabolism disorders, thus overcoming the shortcomings of existing technologies. Summary of the Invention

[0006] To address the aforementioned problems, in a first aspect, this application proposes a compound lactic acid bacteria composition comprising *Lactobacillus plantarum* (accession number GDMCC No. 67514). Lactiplantibacillus plantarum HMPM1006C and Lactobacillus curvilinearis with accession number GDMCC No. 67515 ( Latilactobacillus curvatus HMPM1403C.

[0007] Furthermore, the composition contains *Lactobacillus plantarum* (… Lactiplantibacillus plantarum HMPM1006C and *Lactobacillus curvilinearis* ( Latilactobacillus curvatus The viable count ratio of HMPM1403C is 1:(0.1-3).

[0008] Furthermore, the *Lactobacillus plantarum* ( Lactiplantibacillus plantarum HMPM1006C and *Lactobacillus curvilinearis* ( Latilactobacillus curvatus The live bacteria ratio of HMPM1403C is 1:(0.5-2), for example, it can be 1:0.2, 1:0.5, 1:0.6, 1:1, 1:2, or 1:3.

[0009] Furthermore, the total viable count of the two strains in the compound lactic acid bacteria composition is not less than 1×10⁻⁶. 9 CFU / mL.

[0010] Secondly, this application proposes the use of the aforementioned compound lactic acid bacteria composition in the preparation of products for the prevention and / or adjunctive treatment of metabolic diseases.

[0011] Thirdly, this application proposes the use of the aforementioned compound lactic acid bacteria composition in the preparation of products for the prevention and / or adjunctive treatment of type 2 diabetes.

[0012] Fourthly, this application proposes the use of the aforementioned compound lactic acid bacteria composition in the preparation of products for the prevention and / or adjunctive treatment of obesity.

[0013] Fifthly, this application proposes the use of the aforementioned compound lactic acid bacteria composition in the preparation of products for the prevention and / or adjunctive treatment of hyperlipidemia.

[0014] Fifthly, this application proposes a product for the prevention and / or adjunctive treatment of metabolic diseases, comprising the aforementioned compound lactic acid bacteria composition as an active ingredient.

[0015] Furthermore, the dosage form of the product is a bacterial powder, compressed candy, solid beverage, oral liquid, capsule, or food for special medical purposes.

[0016] Compared with the prior art, this application has the following advantages: The compound composition of *Lactobacillus curvilinearis* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C) proposed in this application achieves synergistic effects through scientific formulation. In terms of blood sugar control, it inhibits α-glucosidase (inhibition rates of 72.62% and 99.87%, respectively) and α-amylase activity, delays carbohydrate absorption, and improves insulin sensitivity, with a blood sugar-lowering effect superior to that of a single strain. In terms of lipid regulation, it promotes cholesterol clearance (clearance rates of 36.11% and 46.01%, respectively), reduces serum lipid levels, and decreases hepatic fat accumulation. Simultaneously, it regulates the balance of inflammatory factors, enhances the body's antioxidant capacity, and both strains exhibit good tolerance to gastrointestinal fluids. *Lactobacillus plantarum* can promote the growth of *Lactobacillus curvilinearis*, enhancing its colonization stability in vivo. This composition has clear efficacy and can be used to prepare various dosage forms such as pharmaceuticals and health foods, providing a natural solution for the prevention and adjuvant treatment of metabolic diseases such as type 2 diabetes and hyperlipidemia, with broad application prospects.

[0017] Regarding its blood sugar-lowering effects, the combination of *Lactobacillus curvilinearis* and *Lactobacillus plantarum* can, on the one hand, reduce postprandial blood glucose peaks by slowing down the absorption rate of carbohydrates in the intestines; and on the other hand, reduce insulin resistance by improving insulin sensitivity in peripheral tissues. The synergistic effect of these two methods achieves comprehensive blood sugar regulation from the "source" to the "end," with significantly better results than the simple superposition of single strains. Regarding its lipid-regulating effects, this invention comprehensively improves common lipid metabolism disorders in type 2 diabetes through a multi-level mechanism. In the combination, the strains promote cholesterol metabolism, thereby effectively reducing serum total cholesterol, triglycerides, and low-density lipoprotein cholesterol levels, helping to reduce the risk of cardiovascular complications in diabetes.

[0018] Other features and advantages of this application will be set forth in the description which follows, and will be apparent in part from the description, or may be learned by practicing the application. The objectives and other advantages of this application may be realized and obtained by means of the structures pointed out in the description, claims and drawings. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 The *Lactobacillus curvilinearis* in Example 1 of this application (… Latilactobacillus curvatus Phylogenetic tree of Lc)HMPM1403C; Figure 2 Lactobacillus plantarum in Example 1 of this application ( Lactiplantibacillus plantarum Phylogenetic tree of Lp)HMPM1006C; Figure 3 The images show colony morphology and Gram staining of *Lactobacillus curvilinearis* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C) in the embodiments of this application. Figure 4A This is a comparison of the growth curves of *Lactobacillus curvilinearis* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C) in the embodiments of this application. Figure 4B This is a comparison of the viable counts of two lactobacillus strains, *Lactobacillus curvilinearis* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C), in the embodiments of this application. Figure 4C This is a comparison of the crystal violet absorbance of *Lactobacillus curvilinearis* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C) in the embodiments of this application. P <0.001 indicates a highly significant difference; Figure 5 The *Lactobacillus curvilinearis* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C) used in the embodiments of this application are effective against high-sugar-induced *C. elegans* (…). C. elegans The improving effect of ) ; where A represents C. elegans The amount of glucose (Glc) in the body, represented by B. C. elegans The body's triglyceride (TG) content, represented by C. C. elegans The total cholesterol (TC) level in the body, where D represents... C. elegans ROS and Nile Red staining, E indicates C. elegans The level of glutathione reductase (GSH) in the body, F represents C. elegans The level of malondialdehyde (MDA) in the body; G indicates... C. elegans Lifespan; different lowercase letters in the diagram (a, b, c, etc.) represent the time between treatments. P The difference was statistically significant at the <0.05 level; Figure 6The figures show the results of oral glucose tolerance test (OGTT), area under the curve (AUC), serum glucose (GLc), fasting blood glucose (FBG), serum insulin, and insulin resistance index (HOMA-IR) in each group of mice after modeling in this application. A represents the oral glucose tolerance test in each group of mice, B represents the change in AUC glucose in each group of mice, C represents the change in serum glucose content in each group of mice, D represents the change in fasting blood glucose in each group of mice, E represents the change in serum insulin content in each group of mice, and F represents the insulin resistance index in each group of mice. Different lowercase letters (a, b, c, etc.) in the figure indicate the time between treatments. P The difference was statistically significant at the <0.05 level; Figure 7 The figures show the expression levels of lipid and inflammatory factor-related indicators (TC, TG, LDL-C, FFA, IL-10, TNF-α) in each group of mice in this application embodiment; where A represents the TC content in the serum of each group of mice at the end of the experiment, B represents the TG content in the serum of each group of mice at the end of the experiment, C represents the LDL-C content in the serum of each group of mice at the end of the experiment, D represents the FFA content in the serum of each group of mice at the end of the experiment, E represents the IL-10 content in the serum of each group of mice at the end of the experiment, and F represents the TNF-α content in the serum of each group of mice at the end of the experiment; different lowercase letters (a, b, c, etc.) in the figure represent the levels between treatments. P The difference was statistically significant at the <0.05 level; Figure 8 The images show pathological sections (H&E staining) of liver and pancreas tissues from each group of mice in the embodiments of this application; where A represents the H&E staining results of liver tissues from each group of mice; and B represents the H&E staining results of pancreas tissues from each group of mice. Figure 9A Figure 1 shows the α-diversity analysis results of each group of mice in the embodiments of this application; where Figure 2a shows the analysis results of the Chao1 index; Figure 3b shows the analysis results of the ACE index; Figure 4c shows the analysis results of the Shannon index; and Figure 5d shows the analysis results of the Simpson index. In the figures, compared with the model group (MC group), P <0.05, P <0.01 indicates a significant or highly significant difference; Figure 9B This is a heatmap showing the correlation between microbial communities and indicators in the embodiments of this application; Figure 9C This is a flower diagram of OTUs / asv of mice in each group in the embodiments of this application; Figure 9D This is a stacked diagram of the phylum-level bacterial community composition of each group of mice in the embodiments of this application; Figure 9EThis is a diagram showing the relative abundance of key bacterial communities at the phylum level in each group of mice in the embodiments of this application; where, diagram a shows the Firmicutes phylum ( Bacillota The relative abundance plot (b) and plot (c) represent the Bacteroidetes phylum ( Bacteroidota Figure c shows the relative abundance of Firmicutes to Bacteroidetes, and Figure d shows the relative abundance of Enterobacter spp. ( Muribaculum Relative abundance plot; in the plot, compared to the model group (MC group), P <0.05, P <0.01, P <0.001 indicates a significant, highly significant, or extremely significant difference; Figure 10A This is a graph showing the glucose metabolism, lipid metabolism, inflammatory factors, and liver enzyme activity indicators of mice in each group in the embodiments of this application; compared with the model group (MC group), P <0.05, P <0.01, P <0.001 indicates a significant, highly significant, or extremely significant difference; Figure 10B These are pathological sections of liver and pancreas tissue from mice in each group in the embodiments of this application (scale bar 50 μm, magnification 400×); compared with the model group (MC group), P <0.05, P <0.01, P <0.001 indicates a significant, highly significant, or extremely significant difference. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0022] Example 1 (I) Strain source and screening: Curved Lactobacillus ( Latilactobacillus curvatus HMPM1403C was obtained by screening chili sauce purchased in Changsha; *Lactobacillus plantarum* ( Lactiplantibacillus plantarum HMPM1006C was selected from pickled cowpeas made at home in Anhui.

[0023] (II) Separation and purification steps: Serial dilution: 0.1 mL of chili sauce and pickled cowpea samples were respectively diluted in 0.9 mL of sterile physiological saline to obtain 10... -1 Dilute the solution, then take 0.1 mL of 10 -1 The diluent was diluted in 0.9 mL of physiological saline to obtain 10 -2 Diluent, follow these steps to obtain 10 -3 10 -4 10 -5 10 -6 Diluent; Coating plate: Take 10 -4 10 -5 10 -6 100 μL of concentration gradient dilution solution was spread onto LBS plates, with one replicate for each gradient.

[0024] Incubation: Place the LBS plates in an anaerobic incubator at 37℃ and incubate for 24-48 h.

[0025] Isolation and purification: Based on the differences in colony morphology, colonies of each morphology were randomly selected and inoculated onto LBS plates using the three-region streak method; single colonies were picked from the streak purification plates for a second streak purification.

[0026] Liquid culture: Pick a single colony from the final purified plate and inoculate it into 10 mL of the corresponding liquid culture medium, and incubate for 12-48 h.

[0027] (III) Strain Identification: The purified strain was subjected to PCR amplification, and the product was detected by agarose gel electrophoresis. Sequencing was performed after a bright single band was observed. The sequencing sequences were aligned using BLAST on NCBI to construct a phylogenetic tree.

[0028] like Figure 1 As shown, the 16S rRNA sequence of *Lactobacillus curvilinearis* (HMPM1403C) is similar to... Latilactobacillus curvatus The type strains showed extremely high homology, clustering into one clade in the phylogenetic tree; for example... Figure 2 As shown, the 16S rRNA sequence of *Lactobacillus plantarum* (HMPM1006C) is similar to... Lactiplantibacillus plantarum The type strains are most closely related, and the clustering results are clear.

[0029] Based on morphological and physiological-biochemical characteristics, the taxonomic positions of the two strains were determined, and they were deposited. The deposit information is as follows: Curved Lactobacillus ( Latilactobacillus curvatusHMPM1403C, depositary institution: Guangdong Provincial Center for Microbial Culture Collection, deposit date: December 23, 2025, accession number: GDMCC No. 67515; Lactobacillus plantarum ( Lactiplantibacillus plantarum HMPM1006C, depositary institution: Guangdong Provincial Center for Microbial Culture Collection, deposit date: December 23, 2025, accession number: GDMCC No.67514.

[0030] (iv) Activation of bacterial strains and preparation of compound bacterial solutions; Strain activation: Take bacterial culture from the preservation tube, inoculate it into MRS liquid medium, and anaerobic incubate at 37°C for 24 h to complete the first generation. Continuous activation is performed for 3 generations until the bacterial culture reaches OD. 600 It remained stable at 1.0 ± 0.1.

[0031] MRS medium formulation: 10.0 g peptone, 8.0 g beef extract, 4.0 g yeast extract, 20.0 g glucose, 2.0 g dipotassium hydrogen phosphate, 2.0 g triammonium citrate, 5.0 g sodium acetate, 0.2 g magnesium sulfate, 0.05 g manganese sulfate, 1.0 mL Tween 80, add water to a final volume of 1 L, and adjust the pH to 6.2 ± 0.2; for solid medium, an additional 15.0 g agar powder needs to be added.

[0032] Preparation of compound bacterial suspensions: The concentrations of the two bacterial suspensions were adjusted to 2 × 10⁻⁶ using the plate coating method for bacterial counting. 9 CFU / mL, mixed at a ratio of *Lactobacillus plantarum* to *Lactobacillus extensively curvedus* of 1:(0.1-3), specifically including 1:1, 1:2, 1:0.5, 1:1.67, and 1:0.6, ensuring a total viable count of not less than 1×10⁻⁶. 9 CFU / mL, collect the supernatant (CFS).

[0033] Example 2 Determination of the hypoglycemic and lipid-lowering abilities and probiotic properties of *Lactobacillus curvilinearis* (HMPM1403C, abbreviated as Lc) and *Lactobacillus plantarum* (HMPM1006C, abbreviated as Lp).

[0034] Fresh bacterial cultures of *Lactobacillus curvilinearis* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C) grown for 24 hours were taken and their OD values ​​were adjusted. 600 Centrifuge at 1.0°C, 4°C, 8000 rpm for 10 minutes, and collect the supernatant (CFS) for various index determinations.

[0035] (1) Determination of α-glucosidase inhibition rate: Mix 150 μL of 0.01M PBS with 75 μL of 5 μM p-nitrophenol-D-glucosidase (PNPG) solution and incubate at 37°C for 10 min. Add 25 μL of CFS and incubate at 37°C for another 10 min. Add 50 μL of 1 U / mL α-glucosidase solution and incubate at 37°C for 10 min. Finally, add 1 mL of 0.1 M Na2CO3 to terminate the reaction and measure the absorbance at 405 nm.

[0036] The inhibition of α-glucosidase activity is calculated according to the following formula: α-glucosidase inhibition rate (%) = [(1-(CD)) / (AB)]×100 Where A is the absorption rate of the mixture without CFS; B is the absorption rate of the mixture without α-glucosidase and CFS; C is the absorption rate of the mixture containing α-glucosidase and CFS; and D is the absorption rate of the mixture without α-glucosidase.

[0037] (2) Determination of α-amylase inhibition rate: 250 μL of CFS was mixed with 250 μL of α-amylase (2 U / mL) and incubated at 25 °C for 10 min. Then, the reaction mixture was reacted with 250 μL of starch solution (1% w / v) at 25 °C for 10 min. The reaction was terminated by adding 500 μL of DNS chromogenic reagent (96 mM DNS and 5.31 M potassium sodium tartrate dissolved in 2 M NaOH solution). The reaction mixture was then boiled for 5 min. After cooling, it was diluted 4-fold with water, and the absorbance was measured at 540 nm.

[0038] The inhibition of α-amylase activity was calculated as follows: α-Amylase inhibition rate (%) = (AB) / A × 100 Where A is the absorbance of the control and B is the absorbance of the sample.

[0039] (3) Cholesterol clearance test: Cholesterol (final concentration 0.2 mg / mL) was added to MRS medium. The activated bacterial strain was adjusted to OD600 = 1 and inoculated into MRS high-cholesterol liquid medium. The culture was incubated in an anaerobic incubator at 37℃ for 24 h. The bacterial suspension was centrifuged at 9000 rpm for 10 min, and the supernatant was collected. The cholesterol content was determined using the o-phthalaldehyde method. o-phthalaldehyde method: 125 μL of the supernatant was added to 50 μL of 1 mg / mL o-phthalaldehyde solution (dissolved in ethanol), shaken thoroughly, and allowed to stand for 10 min. Then, 2 mL of a mixed acid solution (a 1:1 mixture of concentrated sulfuric acid and glacial acetic acid) was added, allowed to stand for 10 min, and the absorbance was measured at 550 nm.

[0040] Cholesterol clearance rate is calculated as follows: Cholesterol clearance rate (%) = (A1-A2) / A1×100, where A1: cholesterol content in the supernatant before fermentation, and A2: cholesterol content in the supernatant after fermentation.

[0041] (4) DPPH· free radical scavenging activity assay: Mix 500 μL of 0.4 mM DPPH, 50 μL of CFS and 900 μL of anhydrous ethanol, react in the dark for 30 min, centrifuge at 8000 rpm for 10 min, remove the supernatant, and measure A1 at 517 nm.

[0042] The DPPH free radical scavenging rate is calculated as follows: DPPH radical scavenging rate (%) = (1-(A1-A2) / A0)×100, where: A0: control group using anhydrous ethanol instead of the test sample solution; A2: blank control group using anhydrous ethanol instead of DPPH solution.

[0043] (5) ABTS + • Free radical scavenging activity assay: A 14 mM ABTS solution was mixed with 4.9 mM K₂S₂O₈ and reacted in the dark for 12 h. The mixture was then diluted with ddH₂O to obtain A. 734nm The absorbance was approximately 2.0. After mixing 20 μL of CFS with 180 μL of ABTS and reacting for 6 min, the absorbance was measured at 734 nm.

[0044] ABTS + The free radical scavenging rate is calculated as follows: ABTS + • Free radical scavenging rate (%) = (1-B / A)×100, where A is the absorbance of the blank control group and B is the absorbance of the sample group.

[0045] (6) Gastrointestinal fluid tolerance test: The bacterial strain was inoculated into MRS medium and cultured at 37°C for 18 h. After washing three times with sterile physiological saline, 500 μL of the bacterial solution was inoculated into 4.5 mL of simulated gastric fluid (pH = 3) and treated at 37°C for 1 h. After treating the sample in gastric fluid at pH = 3 for 3 h, 500 μL of the mixture was added to 4.5 mL of simulated intestinal fluid (pH = 8) and incubated at 37°C for 1 h. After tolerance, the probiotics were sampled and placed in sample wells, MRS wells, and distilled water calibration wells. 100 μL of the bacterial solution was added to 10 μL of CCK-8 reagent, and after incubation at 37°C for 2 h, the absorbance was measured at 450 nm.

[0046] Gastrointestinal fluid tolerance rate is calculated as follows: B = (A (sample well) - A (distilled water)) / (A (MRS well) - A (distilled water)) × 100 Gastric juice tolerance rate (%) = B1 (gastric juice tolerance) / B0 (initial bacterial culture) × 100 Intestinal fluid tolerance rate (%) = B2 (intestinal fluid tolerance) / B1 (gastric fluid tolerance) × 100 The results of each measurement are shown in Table 1: Table 1

[0047] It can be seen that *Lactobacillus curvaturetus* (HMPM1403C) exhibited an α-glucosidase inhibition rate of 72.62 ± 0.14%, an α-amylase inhibition rate of 66.08 ± 0.92%, a cholesterol clearance rate of 36.11 ± 1.78%, and a DPPH inhibition rate of 72.62 ± 0.14%. The clearance rate was 50.73 ± 3.28%, ABTS + The free radical scavenging rate was 56.27 ± 1.74%, the gastric juice tolerance survival rate was 37.39 ± 1.30%, and the intestinal juice tolerance survival rate was 76.47 ± 0.97%. The various indicators of *Lactobacillus plantarum* (HMPM1006C) were 99.87 ± 1.02%, 71.65 ± 1.66%, 46.01 ± 2.66%, 46.18 ± 3.40%, 48.79 ± 1.66%, 40.47 ± 1.78%, and 84.95 ± 16.19%, respectively.

[0048] The CFS of *Lactobacillus curvaturei* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C) with different compound ratios (prepared according to the compound bacterial culture preparation process in Example 1) were tested, along with their inhibition rates on α-glucosidase and α-amylase. The results are shown in Table 2. Table 2

[0049] It can be seen that when the ratio of Lactobacillus plantarum (Lp) to Lactobacillus curvilinearis (Lc) is 1:0.5 and 1:0.67, the inhibition rates of α-glucosidase and α-amylase are both at a high level.

[0050] Example 3 Morphological observation of *Lactobacillus curvilinearis* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C).

[0051] Lactobacillus curvilinearis (HMPM1403C) and Lactobacillus plantarum (HMPM1006C) were inoculated into MRS medium and cultured anaerobically at 37°C for 24 h. The culture was then streaked onto MRS solid medium. Gram staining was used to stained both strains of Lactobacillus.

[0052] Staining conditions as follows Figure 3 As shown, *Lactobacillus curvaturetus* colonies are round and transparent with smooth edges, and turn purple after Gram staining, thus identifying it as a Gram-positive bacterium; *Lactobacillus plantarum* colonies are also round with smooth edges, and turn purple after Gram staining, thus identifying it as a Gram-positive bacterium.

[0053] Example 4 The interaction between *Lactobacillus curvilinearis* (HMPM1403C) and *Lactobacillus plantarum* (HMPM1006C) was investigated and verified.

[0054] (a) Growth curve determination: The two activated bacterial strains were inoculated separately into MRS liquid medium at a 1% inoculation rate (i.e., the inoculated bacterial culture volume accounts for 1% of the total volume of the final culture system) and cultured separately, while also being co-cultured at a 1:1 ratio, with uninoculated MRS liquid medium serving as a blank control. Samples were taken every 2 hours, and the OD value was measured at a wavelength of 600 nm to plot the growth curve.

[0055] like Figure 4A As shown, both strains entered the logarithmic growth phase at 4-12 hours, reached the stationary phase at around 12 hours, and entered the decline phase after 24 hours. The OD of the mixed culture... 600 The nm value was higher than that of Lactobacillus curvilinearis culture alone, indicating that Lactobacillus plantarum can promote the growth of Lactobacillus curvilinearis.

[0056] (ii) Verification of viable cell count: The CFS of one bacterium was mixed with MRS liquid medium and inoculated into the test bacteria at a 1:1 ratio; another bacterium was autoclaved and inoculated into the test bacteria at a 1:1 ratio. After incubation at 37°C for 24 h, the viable count was determined.

[0057] like Figure 4B As shown, after adding the CFS of another strain, the number of viable bacteria in the test strain increased significantly compared with single-strain culture, indicating that the metabolites of the two strains can mutually promote growth.

[0058] (III) Verification by crystal violet staining: Transfer the activated bacterial culture medium to MRS medium at an inoculation concentration of 5% (v / v). Take 200 μL of the diluted suspension and add it to a 96-well plate. Add the same volume of suspension to the compound strain in a 1:1 ratio. After culturing at 37 °C for 24 h, dry it. Add 100 μL of 99% methanol to each well to fix for 15 min. After removing the methanol, add 100 μL of 0.1% crystal violet solution to stain for 30 min. Discard the staining solution and dissolve it with 100 μL of absolute ethanol for 30 min. Measure the absorbance at 570 nm.

[0059] As Figure 4C shown, the average value of OD 570 nm in the Lc group is 0.9, that in the Lp group is 0.8, and that in the Lc+Lp group is 1.9. And ODLc +ODLp < ODLc+Lp, further confirming that there is a mutual promotion effect between the two strains of bacteria.

[0060] Example 5 Establishment of a model of Caenorhabditis elegans ( C. elegans ) induced by high glucose and determination of biochemical indexes.

[0061] (1) Establishment of a model of Caenorhabditis elegans ( C. elegans ) induced by high glucose; Set up 5 experimental groups: 1) Control group (NC group): Use NGM medium; 2) High-glucose group (MC group): Use high-glucose NGM medium containing 4% glucose; 3) Lactobacillus paraplantarum group (Lc group): High-glucose NGM medium (4% glucose) + Lactobacillus paraplantarum bacterial liquid; 4) Lactiplantibacillus plantarum group (Lp group): High-glucose NGM medium (4% glucose) + Lactiplantibacillus plantarum bacterial liquid; 5) Compound group (Lc+Lp group): High-glucose NGM medium (4% glucose) + compound bacterial liquid (Lactobacillus paraplantarum bacterial liquid + Lactiplantibacillus plantarum bacterial liquid).

[0062] Among them, the formula composition of NGM medium is as follows: 3.0 g of sodium chloride, 2.5 g of peptone, 17.0 g of agar, 3.0 g of dipotassium hydrogen phosphate, 2.0 g of potassium dihydrogen phosphate, 5 mg of cholesterol, 1 mM of calcium chloride and 1 mM of magnesium sulfate, add water to 1 L, pH is about 6.0.

[0063] The formula composition of high-glucose NGM medium is as follows: 3.0 g of sodium chloride, 2.5 g of peptone, 17.0 g of agar, 3.0 g of dipotassium hydrogen phosphate, 2.0 g of potassium dihydrogen phosphate, 5 mg of cholesterol, 1 mM of calcium chloride and 1 mM of magnesium sulfate, 40 g of glucose, add water to 1 L, pH is about 6.0.

[0064] (2) Determination of biochemical indicators; 1) TG and TC determination: Caenorhabditis elegans, after intervention to the L4 stage, was collected into 2 mL centrifuge tubes, repeatedly washed with M9 buffer, and lipids were extracted using the Bligh & Dyer method. 500 μL of methanol:chloroform (1:2, v / v) was added to each nematode, and the mixture was ground and crushed. Then, 0.1 mL of 0.85% sodium chloride solution was added, vortexed, and centrifuged at 3000 rpm for 15 min. The lower organic phase was carefully transferred to a 1.8 mL liquid chromatography-tandem chromatography bottle and dried in a vacuum desiccator. 30 μL of Triton X-100 and methanol (1:1, v / v) were added, and the mixture was dried again under vacuum to remove the methanol. Finally, 40 μL of sodium chloride solution was added for analysis. TG and TC were determined using a kit from the Nanjing Jiancheng Biotechnology Institute.

[0065] The results are as follows Figure 5 As shown in BC, the lipid content in the high-glucose group (MC group) was significantly higher than that in the control group (NC group), and probiotic intervention significantly reduced lipid accumulation in Caenorhabditis elegans induced by high glucose. P <0.05).

[0066] 2) GLc, GSH, and MDA assays: Caenorhabditis elegans was collected at the L4 stage after intervention. The samples were repeatedly washed with M9 buffer, resuspended in 300 μL of M9 buffer, and disrupted. The samples were centrifuged at 8000 rpm for 5 min, and the supernatant was collected for analysis. GLc, GSH, and MDA were all measured using a kit from Nanjing Jiancheng.

[0067] like Figure 5 As shown in Figure A, the glucose content in the high-glucose group (MC group) was significantly higher than that in the control group (NC group). Probiotic intervention significantly reduced the glucose content in Caenorhabditis elegans induced by high glucose. P <0.05). For example... Figure 5 As shown in the DF, probiotic intervention can significantly improve the antioxidant capacity of high-sugar-induced Caenorhabditis elegans. P <0.05).

[0068] 3) Nematode lifespan experiment: Nematodes are synchronized, and the eggs are incubated to the L2 stage. The eggs are transferred to a plate every two days until the nematodes die, thus determining the lifespan of the nematodes.

[0069] like Figure 5As shown in Figure G, the average lifespan of each group was as follows: NC group: 16.7 ± 1.2 d, MC group: 10.0 ± 0.0 d, Lc group: 15.3 ± 1.2 d, Lp group: 16.0 ± 0.0 d, and Lc+Lp group: 17.3 ± 1.2 d. The experimental results indicate that probiotic intervention can prolong the lifespan of *C. elegans* induced by high glucose. This *C. elegans* model preliminarily demonstrates the hypoglycemic and lipid-lowering abilities of probiotics.

[0070] Example 6 Establishment and grouping design of type 2 diabetes mouse model.

[0071] (1) Preparation of bacterial culture: The activated bacterial suspension was centrifuged at 4°C and 8000 rpm for 10 min, washed three times with physiological saline, and the bacterial concentration was adjusted to 2 × 10⁻⁶. 8 CFU / mL, used for subsequent experiments.

[0072] (2) Establishment of a type 2 diabetes mouse model: The experimental animals were 6-week-old male ICR mice (20-25 g), housed in the animal facility of the College of Life Sciences and Medical Engineering, Anhui University (ethics review number: IACUC (AHU)-2025-097). A 12-hour / 12-hour light-dark cycle was used, with suitable temperature and humidity. Mice had free access to food and water. After one week of acclimatization, the mice were randomly assigned to groups of 8 mice each, with 4 mice per cage.

[0073] Methods for inducing type 2 diabetes: After feeding on a high-fat diet for 4 weeks, streptozotocin (STZ) was injected intraperitoneally to induce the diabetes.

[0074] (3) Animal experiment grouping: 1) Normal group (NC group): fed normal maintenance diet for 4 weeks, and gavage 200 μL of physiological saline daily; in the 5th week, intraperitoneal injection of an equal volume of 0.1 mmol / L citrate buffer for 5 consecutive days; gavage 200 μL of physiological saline daily from the 5th to the 8th week until the end of the experiment.

[0075] 2) Model group (MC group): fed with high-fat diet for 4 weeks, and gavaged with 200 μL of physiological saline daily; in the 5th week, streptozotocin (STZ) was injected intraperitoneally for 5 consecutive days at a dose of 50 mg / kg body weight; if blood glucose no longer returned to normal level after 72 h (blood glucose ≥11.1 mmol / L), it was considered that the model was successfully established; gavaged with 200 μL of physiological saline daily from the 5th to the 8th week until the end of the experiment.

[0076] 3) Lactobacillus curvatureensis group (Lc group): fed a high-fat diet for 4 weeks, and administered 2×10⁻⁶ Lactobacillus via gavage daily. 8CFU suspension of *Lactobacillus rubrum*; intraperitoneal injection of STZ at a dose of 50 mg / kg body weight for 5 consecutive days during week 5; successful model establishment is considered when blood glucose does not return to normal levels after 72 hours; 2×10 gavage daily during weeks 5-8. 8 CFU was applied to a wide range of Lactobacillus suspensions until the end of the experiment.

[0077] 4) Lactobacillus plantarum group (Lp group): fed with a high-fat diet for 4 weeks, and administered 2×10 gavages daily. 8 CFU of *Lactobacillus plantarum* suspension; intraperitoneal injection of STZ at a dose of 50 mg / kg body weight for 5 consecutive days during week 5; successful model establishment is considered when blood glucose no longer returns to normal levels after 72 hours; 2×10 mg / kg body weight via gavage daily from week 5 to week 8. 8 The CFU (Chemical Lactobacillus plantarum) suspension was used until the end of the experiment.

[0078] 5) Compound group (Lc+Lp group): fed with high-fat diet for 4 weeks, with 2×10 gavages daily. 8 A bacterial suspension co-cultured with two strains of Lactobacillus in CFU; STZ was injected intraperitoneally at a dose of 50 mg / kg body weight for 5 consecutive days during week 5; successful modeling was considered when blood glucose no longer returned to normal levels after 72 hours; 2×10 mg / kg body weight was administered by gavage daily from week 5 to week 8. 8 The two strains of lactobacillus in CFU were co-cultured in suspension until the end of the experiment.

[0079] 6) Positive drug group (Met group): fed with high-fat diet for 4 weeks, and administered 200 μL of physiological saline by gavage daily; in the 5th week, STZ was injected intraperitoneally at a dose of 50 mg / kg body weight for 5 consecutive days; if blood glucose no longer returned to normal level after 72 h, the model was considered successful; from the 5th to the 8th week, metformin (Met) was administered by gavage daily at a dose of 100 mg / kg body weight until the end of the experiment.

[0080] Example 7 Mouse serum, formed fecal samples, and tissue samples were collected.

[0081] Before the end of the experiment, the mice were fasted but allowed free access to water for 12 hours. Blood was collected through the orbital sinus, incubated on ice for 2 hours, centrifuged at 3500 r / min for 15 min, and the supernatant was collected to obtain serum, which was stored at -20℃ for subsequent physiological and biochemical and inflammatory factor index detection.

[0082] After blood was collected from mice, the contents of the liver, pancreas, colon, and cecum were collected and processed into different tissues: a portion of the tissue was cut into pieces and fixed in a pre-prepared 4% paraformaldehyde solution for histopathological observation; the remaining tissues were flash-frozen in liquid nitrogen and then stored at -80°C.

[0083] Example 8 Oral glucose tolerance test analysis, fasting blood glucose in mice, biochemical indicators and inflammatory factors determination.

[0084] (1) Oral glucose tolerance test (OGTT): One week before the end of the animal experiment, mice were given a glucose solution of 2 g / kg body weight by gavage. Blood glucose was measured at 0 min, 30 min, 60 min, 90 min and 120 min, and the area under the curve (AUC) was calculated to assess the oral glucose tolerance of the mice.

[0085] like Figure 6 As shown in Figure AB, the blood glucose levels in the normal group (NC group) remained unchanged, while the blood glucose levels in the model group (MC group) were significantly higher than those in the normal group (NC group). P <0.05%. After probiotic intervention, oral glucose tolerance in type 2 diabetic mice was significantly restored ( P <0.05), indicating that *Lactobacillus curvilinearis*, *Lactobacillus plantarum*, and the combined strains can all improve glucose tolerance and symptoms of type 2 diabetes in mice.

[0086] (2) Measurement of fasting blood glucose (FBG) in mice: After successful modeling, fasting blood glucose was measured weekly. Mice were fasted for 12 hours but allowed free access to water, and blood was collected from the tail tip using a blood glucose meter. Glucose (GLc) was measured using a Nanjing Jiancheng reagent kit, and insulin (Insulin) was measured using a Ruixin Bio ELISA kit. The insulin resistance index (HOMA-IR) was calculated.

[0087] like Figure 6 As shown in Figure D, two weeks after STZ injection, there was no significant difference in blood glucose levels among the groups in mice. P >0.05); Starting from week 7, the fasting blood glucose levels of the Lactobacillus curvilinearis group (Lc group), Lactobacillus plantarum group (Lp group), and the compound group (Lc+Lp group) were significantly lower than those of the model group (MC group), indicating that probiotic intervention can reduce blood glucose in type 2 diabetic mice.

[0088] like Figure 6 As shown in Figure C, the serum glucose level in the model group (MC group) was significantly higher than that in the normal group (NC group). P <0.05), probiotic intervention significantly reduced serum glucose levels ( P <0.05).

[0089] like Figure 6 As shown in Figure E, the serum insulin levels in the model group (MC group) were significantly higher than those in the normal group (NC group). P <0.05, there was no significant difference between the Lc group and the model group (MC group). P>0.05), while the compound group (Lc+Lp group) was significantly lower than the model group (MC group) ( P <0.05). The Homeostasis Model of Insulin (HOMA-IR) index was calculated by measuring fasting blood glucose (FBG) and fasting insulin levels to assess the degree of insulin resistance in the body. The results are as follows: Figure 6 As shown in Figure F, the HOMA-IR index of the model group (MC group) was significantly higher than that of the normal group (NC group), indicating that T2DM mice exhibited typical insulin resistance. However, the HOMA-IR index was significantly restored after probiotic intervention, especially in the compound group (Lc+Lp group), indicating that probiotic intervention can improve insulin resistance caused by type 2 diabetes.

[0090] (3) Biochemical index determination: Triglycerides (TG), total cholesterol (TC), and free fatty acids (FFA) were measured using the Nanjing Jiancheng kit, while interleukin-10 (IL-10) and tumor necrosis factor (TNF-α) were measured using the Ruixin Bio ELISA kit.

[0091] like Figure 7 As shown in the AD diagram, the serum lipid levels in the model group (MC group) were significantly higher than those in the probiotic intervention group ( P <0.05); such as Figure 7 As shown in Figure E, the IL-10 content in the model group (MC group) was significantly lower than that in the normal group (NC group). P <0.05%, probiotic intervention can significantly increase IL-10 levels ( P <0.05); such as Figure 7 As shown in Figure F, the TNF-α level in the model group (MC group) was significantly higher than that in the normal group (NC group), and probiotic intervention could significantly reverse this change. P <0.05).

[0092] From the perspective of glucose and lipid metabolism indicators, probiotic intervention can effectively improve type 2 diabetes.

[0093] (4) Observation of histopathological sections of mouse liver and pancreas tissue: Mouse liver and pancreas tissue samples were sent to Wuhan Sewell Biotechnology Co., Ltd. for H&E staining analysis and microscopic observation.

[0094] like Figure 8 As shown in Figure A, the hepatocytes of mice in the normal group (NC group) were arranged tightly and neatly, while the hepatocytes of mice in the model group (MC group) showed fatty vacuoles and irregular cell morphology; after probiotic intervention, hepatocyte damage was significantly improved. Figure 8As shown in Figure B, the pancreatic islets of the normal group (NC group) mice were round with clear boundaries and abundant and evenly distributed β cells; the pancreatic islets of the type 2 diabetic mice (MC group) were atrophied, irregular in shape, with amyloid deposition, reduced β cells and vacuolation; probiotic intervention can significantly improve pancreatic tissue damage.

[0095] Example 9 Analysis of gut microbiota diversity using 16S rRNA from mouse feces.

[0096] Mouse fecal samples were sent to Shanghai Lingen Biotechnology Co., Ltd. for sequencing. The sequencing results were analyzed via the website http: / / www.cloud.biomicroclass.com / CloudPlatform / home.

[0097] like Figure 9A As shown in the figure, α-diversity analysis revealed that the gut microbiota diversity in the model group (MC group) was significantly lower than that in the normal group (NC group), while probiotic intervention significantly restored both the diversity and richness of the gut microbiota. P <0.05).

[0098] like Figure 9B As shown, the correlation analysis between groups indicates that... Muribaculum It showed a negative correlation with INS and TNF-α, and a positive correlation with IL-10, suggesting... Muribaculum It may have potential effects in improving type 2 diabetes mellitus and reducing inflammation.

[0099] like Figure 9C As shown, a flower plot was constructed based on the presence / absence data of OTUs / asv to visually display the distribution of shared and unique microbial taxa in the samples. Additional petals represent the number of unique taxa for each sample. Compared to the model group (MC group), the normal group (NC group) showed a higher sample-specific OTU / asv count. Sample specificity can be improved through probiotic intervention.

[0100] like Figure 9D Phylogenetic analysis showed that the model group (MC group) exhibited significant ecological imbalance compared to the normal group (NC group), characterized by: Bacteroidota The relative abundance of it decreased significantly.

[0101] like Figure 9E As shown in the AD diagram, at the phylum level, the model group (MC group) is Bacteroidetes phylum ( Bacteroidota The abundance of Firmicutes was lower than that of the normal group (NC group). Bacillota The abundance increased significantly. P <0.05), the ratio of relative abundance of Firmicutes and Bacteroidetes was significantly higher than that of the normal group (NC group).P <0.05%, at the genus level, Enterobacter ( Muribaculum Significantly reduced ( P (<0.05), probiotic intervention significantly reversed this change. These results indicate that probiotic intervention can improve the gut microbiota composition of type 2 diabetic mice.

[0102] Example 10 Establishment and grouping design of hyperglycemic mouse model.

[0103] (1) Preparation of bacterial suspension: The activated bacterial suspension was centrifuged at 8000 rpm for 10 min at 4℃, washed 3 times with physiological saline, and then the bacterial concentration was adjusted to 1×10⁻⁶. 9 CFU / mL was used for subsequent experiments.

[0104] (2) Establishment of a hyperglycemic mouse model. Experimental animals: Six-week-old male ICR mice (20-25 g) were housed in the animal facility of the College of Life Sciences and Medical Engineering, Anhui University (ethics review number: IACUC(AHU)-2025-097). A 12h / 12h light-dark alternation environment was used, with suitable temperature and humidity, and free access to food and water. After one week of acclimatization, the formal experiment was conducted, and the mice were randomly divided into groups of 8 mice each, with 4 mice per cage.

[0105] One method for creating a hyperglycemic model involved feeding the animal with a high-fat diet and administering 1 g / kg body weight of ethanol (20%, v / v) via gavage daily.

[0106] (3) Animal experiment grouping.

[0107] 1) Normal group (NC group): fed normal maintenance feed for 12 weeks, and gavaged with 200 μL of physiological saline every day.

[0108] 2) Model group (MC group): fed a high-fat diet for 12 weeks, and administered 200 μL of physiological saline by gavage daily, and ethanol by gavage at a dose of 1g / kg body weight daily until the end of the experiment.

[0109] 3) Lactobacillus curvatureensis group (Lc group): fed a high-fat diet for 12 weeks, with 2×10⁻⁶ gavage daily. 8 CFU of *Lactobacillus flexuralis* suspension was administered by gavage at a dose of 1 g / kg of ethanol daily until the end of the experiment.

[0110] 4) Lactobacillus plantarum group (Lp group): fed a high-fat diet for 12 weeks, and administered 2×10⁶ gavage daily. 8 CFU of *Lactobacillus plantarum* suspension was administered by gavage daily at a dose of 1 g / kg body weight of ethanol until the end of the experiment.

[0111] 5) Combined Lactobacillus curvilinearis and Lactobacillus plantarum (Lc+Lp group): fed a high-fat diet for 12 weeks, with 2×10⁻⁶ gavage daily. 8 The bacterial suspension of two strains of lactobacillus co-cultured in CFU was administered by gavage with ethanol at a dose of 1 g / kg body weight daily until the end of the experiment.

[0112] (4) Analysis of relevant indicators.

[0113] like Figure 10A As shown in the AC figure, the fasting blood glucose, serum insulin and HOMA-IR index of the model group (MC group) were significantly increased compared with the normal group (NC group), and these indicators were significantly reduced after probiotic intervention.

[0114] like Figure 10B As shown, histopathological analysis revealed that, similar to the damage induced by STZ, alcohol caused significant damage to metabolic organs. In this model, probiotics also effectively protected the area and structure of the pancreatic islets. In the liver, alcohol induced severe alveolar steatosis, a phenotype distinct from but equally severe steatosis induced by HFD. Lc+Lp alleviated this damage, consistent with the effective hepatoprotective effect observed in the STZ / HFD model. Figure 10A As shown in the dg plot, the alcohol model also reproduced key metabolic disorders of T2DM. Serum TC, TG, and LDL-C levels were elevated in the model group (MC group), and significantly returned to normal after probiotic intervention. The systemic pro-inflammatory state indicated by elevated serum TNF-α was effectively suppressed by Lp and Lc+Lp. This dual improvement in lipid metabolism and inflammation is similar to the core treatment outcome in STZ / HFD diabetic mice. Unlike the STZ / HFD model, we investigated the specific mechanisms of alcohol toxicity. Alcohol feeding significantly inhibited the activity of key hepatic enzymes for alcohol detoxification, ADH and ALDH. Figure 10A As shown in the figure, Lp and Lc+Lp treatments significantly restored ADH activity. However, the degradation rate of ALDH did not show a significant difference. This new finding suggests an additional etiology-specific mechanism in which probiotics may enhance the clearance of ethanol and its toxic metabolite acetaldehyde, thereby mitigating direct hepatotoxicity and metabolic damage.

[0115] Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application.

Claims

1. A compound lactic acid bacteria composition, characterized in that, Contains *Lactobacillus plantarum* with accession number GDMCC No. 67514. Lactiplantibacillus plantarum HMPM1006C and Lactobacillus curvilinearis with accession number GDMCC No. 67515 ( Latilactobacillus curvatus HMPM1403C.

2. The compound lactic acid bacteria composition according to claim 1, characterized in that, The composition contains Lactobacillus plantarum ( Lactiplantibacillus plantarum HMPM1006C and *Lactobacillus curvilinearis* ( Latilactobacillus curvatus The viable count ratio of HMPM1403C is 1:(0.1-3).

3. The compound lactic acid bacteria composition according to claim 1, characterized in that, The plant lactobacillus ( Lactiplantibacillus plantarum HMPM1006C and *Lactobacillus curvilinearis* ( Latilactobacillus curvatus The viable count ratio of HMPM1403C is 1:(0.5-2).

4. The compound lactic acid bacteria composition according to any one of claims 1-3, characterized in that, The total viable count of the two strains in the compound lactic acid bacteria composition is not less than 1×10⁻⁶. 9 CFU / mL.

5. The use of the compound lactic acid bacteria composition according to any one of claims 1-3 in the preparation of products for the prevention and / or adjunctive treatment of metabolic diseases.

6. The use of the compound lactic acid bacteria composition according to any one of claims 1-3 in the preparation of products for the prevention and / or adjunctive treatment of type 2 diabetes.

7. The use of the compound lactic acid bacteria composition according to any one of claims 1-3 in the preparation of products for the prevention and / or adjunctive treatment of obesity.

8. The use of the compound lactic acid bacteria composition according to any one of claims 1-3 in the preparation of products for the prevention and / or adjunctive treatment of hyperlipidemia.

9. A product for the prevention and / or adjunctive treatment of metabolic diseases, characterized in that, The compound lactic acid bacteria composition according to any one of claims 1-3 is included as an active ingredient.

10. The product according to claim 9, characterized in that, The dosage form of the product is bacterial powder, compressed candy, solid beverage, oral liquid, capsule, or food for special medical purposes.

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