Uses of probiotics in the preparation of hypoglycemic components

Abstract

This invention provides the use of a probiotic composition for preparing a hypoglycemic agent. The probiotic composition contains a *Lactobacillus helveticus* strain UA881, obtained from the Patent Microbial Host Center (NPMD) of the Japan Product Evaluation Technology Base, with accession number NITE BP-03802. The effective dose of the *Lactobacillus helveticus* strain UA881 in the probiotic composition is 4 × 10⁹ CFU / day to 6 × 10⁹ CFU / day. This hypoglycemic agent is administered to obese patients. This invention selects specific types and doses of probiotic strains, enabling the probiotic composition to be used to prepare a hypoglycemic agent, thereby lowering blood sugar in obese patients using this agent and achieving the purpose of blood sugar regulation.

Description

Technical Field

[0001] This invention relates to the use of a probiotic, and more particularly to the use of a probiotic in the preparation of a hypoglycemic composition. Prior Technology

[0002] With social development and technological progress, the global obesity rate has been rising year by year in recent years, making obesity one of the common but easily overlooked chronic diseases, and its impact and harm to human health should not be ignored.

[0003] The main cause of obesity is usually a sustained excess of calorie intake compared to calorie expenditure. This causes excess calories to be converted into fat and accumulate in the body, resulting not only in direct weight gain and increased body fat, but also indirectly in disrupting fat cell function, leading to metabolic imbalances and chronic inflammation, thus negatively impacting health. Furthermore, obesity can also raise blood lipids and blood sugar levels, increasing the risk of other diseases such as cardiovascular disease, diabetes, hypertension, or even cancer.

[0004] When blood sugar levels rise, the pancreas secretes insulin to convert glucose into glycogen, thus lowering blood sugar. However, prolonged high blood sugar can gradually reduce the body's ability to respond to insulin, eventually leading to insulin resistance (IR), which can result in the body being unable to lower blood sugar even with insulin. This can ultimately lead to type 2 diabetes, a difficult-to-treat condition. Therefore, controlling blood sugar in its early stages to prevent prolonged high blood sugar levels can effectively reduce the risk of developing diabetes later in life.

[0005] Therefore, regulating blood sugar to reduce the risk of subsequent diseases has become a key research focus in related technological fields. While current research has confirmed the potential of probiotics in controlling obesity, regulating blood sugar, and improving metabolic diseases, the persistently high obesity rate and the resulting increasing demand for blood sugar regulation necessitate further research and development of new technologies or products capable of regulating and lowering blood sugar, in order to provide more solutions to the challenge of blood sugar control. Summary of the Invention

[0006] In view of the existing need for blood glucose regulation, the purpose of this invention is to provide an application of a probiotic composition, that is, the probiotic composition can exhibit the effect of lowering blood glucose when administered to obese patients, and thus has the potential to be further used as a product or treatment for blood glucose regulation.

[0007] To achieve the aforementioned objectives, the present invention provides the use of a probiotic composition for preparing a hypoglycemic agent, wherein the probiotic composition comprises a *Lactobacillus helveticus* UA881 strain, which is obtained from the Patent Microorganisms Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE) in Japan, with the registration number NITE BP-03802; the effective dose of the *Lactobacillus helveticus* UA881 strain in the probiotic composition is 4 × 10⁹ CFU / day to 6 × 10⁹ CFU / day; and the hypoglycemic agent is administered to obese patients.

[0008] This invention utilizes specific types and dosages of probiotic strains to enable the probiotic composition to be used in the preparation of a blood sugar-lowering component, thereby lowering blood sugar levels in obese patients using the blood sugar-lowering component and achieving the purpose of regulating blood sugar.

[0009] According to the present invention, "CFU" is an abbreviation for colony-forming unit, representing the number of viable bacterial colonies that a sample can generate on a culture medium.

[0010] In some embodiments of the present invention, the probiotic composition comprises a metabolite of the Lactobacillus helveticus UA881 strain.

[0011] In some embodiments of the present invention, the *Lactobacillus helveticus* UA881 strain is in a viable, inactive, or a combination thereof. In other embodiments of the present invention, the *Lactobacillus helveticus* UA881 strain is in a viable state.

[0012] In some embodiments of the present invention, the probiotic composition may also be used to prepare compositions for reducing weight, body mass index (BMI), and / or waist circumference.

[0013] In some embodiments of the present invention, the probiotic composition may also be used to prepare compositions that reduce visceral fat levels and / or fecal lipid content.

[0014] In some embodiments of the present invention, the probiotic composition may also be used to prepare compositions that enhance the abundance of intestinal flora.

[0015] In some embodiments of the present invention, the probiotic composition may also be used to prepare a composition that increases the butyric acid content in feces.

[0016] In some embodiments of the present invention, the probiotic composition may also be used to prepare compositions for increasing the number of Bacteroidetes bacteria in the gut.

[0017] In some embodiments of the present invention, the probiotic composition may also be used to prepare compositions that reduce the ratio (F / B ratio) between Firmicutes and Bacteroidetes in the gut.

[0018] In some embodiments of the present invention, the probiotic composition may also be used to prepare a composition for increasing the number of probiotics in the gut.

[0019] In some embodiments of the present invention, the probiotic composition may also be used to prepare compositions that increase the number of anaerobic bacteria, Prevotella bacteria and / or Broutella bacteria in the gut.

[0020] In some embodiments of the present invention, the obese patient refers to a male with a body fat percentage greater than 25% or a BMI between 25 kg / m² and 27 kg / m²; or a female with a body fat percentage greater than 30% or a BMI between 25 kg / m² and 27 kg / m².

[0021] In some embodiments of the present invention, the hypoglycemic component is administered to patients with a serum total cholesterol level greater than 200 mg / dL.

[0022] In some embodiments of the present invention, the blood sugar-lowering component may be administered once daily, but is not limited thereto.

[0023] In some embodiments of the present invention, the blood glucose-lowering component is a food component or a pharmaceutical component.

[0024] In some embodiments of the present invention, the food composition may be a general food, nutritional supplement, health food, food additive, or a combination thereof, but is not limited thereto.

[0025] In some embodiments of the present invention, the pharmaceutical composition may be a drug, but is not limited thereto.

[0026] In some embodiments of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or additive. The pharmaceutically acceptable carrier or additive may comprise, but is not limited to, a solvent, buffer, emulsifier, suspending agent, decomposer, disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, wetting agent, lubricant, absorption delaying agent, liposome, surfactant, or other similar or suitable carriers or additives of the present invention, or combinations thereof.

[0027] In some embodiments of the present invention, the solvent comprises water, normal saline, phosphate buffered saline (PBS), alcohol, aqueous solution containing alcohol, or combinations thereof, but is not limited thereto.

[0028] In some embodiments of the present invention, the pharmaceutical composition is an enteral dosage form. The enteral dosage form includes, but is not limited to, solutions, suspensions, powders, tablets, pills, syrups, lozenges or troche, chewing gum, capsules, or other similar or applicable enteral dosage forms of the present invention, or combinations thereof.

[0029] In some embodiments of the present invention, the enteral dosage form is an oral dosage form.

[0030] According to the present invention, the "effective dose" refers to the dose required to produce the desired biological response, that is, the dose required to produce the therapeutic effect or the desired effect.

[0031] In this specification, the range indicated by "smallest value to largest value" means, unless otherwise specified, that the range is greater than or equal to the smallest value and less than or equal to the largest value. For example, an effective dose of 4×10⁹ CFU / day to 6×10⁹ CFU / day means that the effective dose range is "greater than or equal to 4×10⁹ CFU / day and less than or equal to 6×10⁹ CFU / day". Simple Explanation of the Diagram

[0032] Figures 1A to 1C show the results of body weight, BMI, and waist circumference measured on day 0 and day 28, respectively, under the condition of taking the sample of Control Example 1 or Example 1. Figures 2A and 2B show the results of visceral fat grade and fecal lipid content measured on day 0 and day 28, respectively, under the condition of taking the sample of Control Example 1 or Example 1. Figures 3A and 3B show the results of serum triglyceride levels measured on day 0 and day 28, respectively, under the condition of taking the sample of Control Example 1 or Example 1. Figures 4A to 4C show the results of serum total cholesterol levels on day 0 and day 28, and serum LDL-C levels in subjects with serum total cholesterol levels greater than 200 mg / dL, under the condition of taking the sample of Control Example 1 or Example 1. Figure 5 shows the results of pre-meal serum glucose levels measured on day 0 and day 28 in subjects with serum total cholesterol levels greater than 200 mg / dL, under the condition of taking the sample of Control Example 1 or Example 1. Figure 6 shows the results of serum uric acid levels measured on day 0 and day 28 in male subjects with serum uric acid levels greater than 7 mg / dL and female subjects with serum uric acid levels greater than 6 mg / dL, under the condition of taking the sample of Control Example 1 or Example 1. Figures 7A to 7H show the results of measuring OTUs, ACE, fecal butyrate content, number of Bacteroidetes, F / B ratio, bacterial ratio of Anaerobic Bacteria, bacterial ratio of Prevotella, and bacterial ratio of Broutella on day 0 and day 28, respectively, under the condition of taking the sample of Control Example 1 or Example 1. Implementation

[0033] The following examples and comparative examples are provided to illustrate the implementation of the present invention. Those skilled in the art can easily understand the advantages and effects of the present invention through the contents of this specification, and can make various modifications and changes without departing from the spirit of the present invention to implement or apply the contents of the present invention.

[0034] [Example] [1] Lactobacillus helveticus [UA881] [Strain sample] [ ]

[0035] After obtaining Lactobacillus helveticus strain UA881 with the accession number NITE BP-03802 from NPMD, it was inoculated into MRS medium (De Man-Rogosa-Sharpe medium, purchased from BD Difco™) at a concentration of approximately 1% (i.e., approximately 1×10⁶ CFU / mL) and cultured at 37°C for approximately 18 to 24 hours to obtain Lactobacillus helveticus strain UA881.

[0036] Next, the *Lactobacillus helveticus* UA881 bacterial culture was inoculated at a concentration of approximately 5% (approximately 5 × 10⁶ CFU / mL) into approximately 4 liters of YPG fermentation medium (placed in a 5-liter fermentation tank, containing yeast extract, soy peptone, and glucose, hence abbreviated as YPG), and fermented at 37°C, a stirring speed of 100 rpm, and an aeration rate of 3 liters / minute for approximately 4 to 8 hours. Subsequently, the bacterial culture was inoculated at a concentration of approximately 6% (approximately 5 × 10⁶ CFU / mL) into approximately 180 liters of YPG fermentation medium (placed in a 250-liter fermentation tank), and fermented at 37°C and a stirring speed of 60 rpm. Fermentation was carried out for approximately 4 to 8 hours at a stirring speed of 30 rpm, an aeration rate of 30 liters / minute, and a tank pressure of 1 kg / cm². Subsequently, the bacterial culture was inoculated at a concentration of approximately 10% (approximately 1 × 10⁷ CFU / mL) into approximately 1800 liters of YPG fermentation medium (placed in a 2500 liter fermentation tank), and fermented for approximately 10 to 12 hours at a temperature of 37°C, a stirring speed of 60 rpm, an aeration rate of 250 liters / minute, and a tank pressure of 1 kg / cm² to complete the fermentation.

[0037] Finally, the fermented bacterial solution was centrifuged, freeze-dried and homogenized to obtain live bacterial powder of Lactobacillus helveticus UA881 strain, which is the Lactobacillus helveticus UA881 strain sample of Example 1.

[0038] [Comparison Example] [1]

[0039] The sample in Control Example 1 was made of maltodextrin, meaning that the sample in Control Example 1 did not contain the Lactobacillus helveticus UA881 strain.

[0040] [Experimental Example] [1] [Human clinical trials] [ ]

[0041] Trial Example 1 was conducted as a randomized, double-blind, placebo-controlled study and was approved by the Human Trial Ethics Committee of Chung Shan Medical University Hospital. It was also registered on the clinical trial platform of the National Institutes of Health in the United States.

[0042] Specifically, this trial recruited 50 participants aged 20 to 65 years with a BMI between 25 kg / m2 and 27 kg / m2, and a body fat percentage greater than 25% for males or greater than 30% for females (considered as obese patients). They were randomly assigned to the probiotic group (i.e., receiving the Lactobacillus helveticus UA881 strain sample from Example 1, hereinafter referred to as Example 1 for this group) totaling 25 participants, and the placebo group (i.e., receiving the sample from Control Example 1, hereinafter referred to as Control Example 1 for this group) totaling 25 participants. The dose of the Lactobacillus helveticus UA881 strain sample from Example 1 was 5 × 10⁹ CFU / day.

[0043] Subjects in each group took either the sample from Control Example 1 or the Lactobacillus helveticus UA881 strain from Example 1 orally every morning on an empty stomach for 4 weeks (28 days in total). The following parameters were measured before administration (day 0) and after 4 weeks (day 28): weight, BMI, waist circumference, visceral fat level, fecal lipids, serum triglycerides, total cholesterol, low-density lipoprotein cholesterol (LDL-C), pre-meal serum glucose (Glucose AC), serum uric acid, and gut microbiota-related indicators. The gut microbiota-related indicators include: operational taxonomic units (OTUs), abundance-based coverage estimator (ACE), fecal butyric acid, Bacteroidetes, F / B ratio (the ratio between Firmicutes and Bacteroidetes, hence simply called F / B ratio), Anaerostipes spp., Prevotella spp., and Blautia spp.

[0044] The measurement results of the above items are shown in Figures 1A to 7H. In each experimental chart, the left-hand result (light gray) of Control Example 1 or Example 1 is the measurement result before sample administration (i.e., day 0), and the right-hand result (medium gray) is the measurement result after 4 weeks of continuous sample administration (i.e., day 28). In addition, the number of subjects who underwent measurement for different items is listed below, denoted by n. The results of day 0 and day 28 were statistically analyzed using a paired-sample t-test to determine whether there was a significant difference between them; where p-values ​​less than 0.05 were marked with "*", p-values ​​less than 0.01 were marked with "**", and p-values ​​less than 0.001 were marked with "***".

[0045] [(1)] [weight,] [BMI] [Waist circumference] [ ]

[0046] The results of the weight, BMI and waist circumference of the subjects in Comparative Example 1 and Example 1 are shown in Figure 1A, Figure 1B and Figure 1C, respectively.

[0047] As can be seen from the results in Figures 1A to 1C, the results of Example 1 on day 28 were significantly lower than those on day 0 in terms of weight, BMI, and waist circumference. Furthermore, Example 1 also showed a greater reduction in weight, BMI, and waist circumference than Control Example 1. Therefore, it can be concluded that continuous administration of the Lactobacillus helveticus UA881 strain sample from Example 1 for 4 weeks indeed demonstrated an effect in reducing weight, BMI, and waist circumference.

[0048] [(2)] [Visceral fat grade and fecal lipids]

[0049] The results of visceral fat grade and fecal lipid content obtained by the subjects of Control Example 1 and Example 1 are shown in Figure 2A and Figure 2B, respectively. Visceral fat grade was determined using an InBody 270 body composition analyzer (manufactured by Biospace Co., Ltd., South Korea). Fecal lipid content was determined as follows: 0.5 g of fecal sample was placed in a 15 mL centrifuge tube, 7 mL of physiological saline was added and shaken well, followed by 7 mL of Folch solution (a 2:1 mixture of chloroform and methanol) and shaken well. The tube was centrifuged at 2000 times gravity (×g) for 10 minutes at room temperature. After centrifugation, the lower organic layer was collected using a needle into a glass tube. The glass tube was placed in a 45°C water bath and dried with nitrogen. 1 mL of isopropanol was then added, the tube was sealed with paraffin film, and the tube was reconstituted using an ultrasonic shaker to obtain a fecal lipid extract. Finally, the lipid content was determined using TRIGLYCERIDES Liquid (purchased from Sentinel Diagnostics, model REF 17624H) with a CANON-TBATM-120FR automated biochemical analyzer.

[0050] As can be seen from the results in Figures 2A and 2B, the results of Example 1 on day 28 were significantly lower than those on day 0, regardless of visceral fat grade or fecal lipids. Furthermore, Example 1 also showed a greater reduction in both visceral fat grade and fecal lipids than Control Example 1. Therefore, it can be concluded that continuous administration of the *Lactobacillus helveticus* UA881 strain sample from Example 1 for 4 weeks indeed demonstrated an effect in reducing visceral fat grade and fecal lipid content.

[0051] [(3)] [Serum triglycerides]

[0052] The serum triglyceride levels of subjects in Control Example 1 and Example 1 are shown in Figures 3A and 3B, respectively; Figure 3B shows the results of screening subjects with serum triglyceride levels greater than 100 mg / dL. The serum triglyceride level was determined by centrifuging the blood sample at 3000×g for 15 minutes, collecting the supernatant serum, and analyzing it using a Cobasproe 801 fully automated chemiluminescence immunoassay analyzer to obtain the serum triglyceride level.

[0053] As shown in Figure 3A, the serum triglyceride level of Example 1 on day 28 was significantly lower than that on day 0. Furthermore, Figure 3B shows that Example 1 demonstrated a better effect in reducing serum triglyceride levels in subjects with elevated levels. On the other hand, as shown in Figures 3A and 3B, the serum triglyceride level of Control Example 1 was higher on day 28 than on day 0, indicating that it did not have the effect of reducing serum triglyceride levels. Therefore, it can be concluded that continuous administration of the *Lactobacillus helveticus* UA881 strain sample from Example 1 for 4 weeks indeed demonstrated an effect in reducing serum triglyceride levels.

[0054] [(4)] [Serum total cholesterol and] [LDL-C]

[0055] The serum total cholesterol and LDL-C levels of the subjects in Comparative Example 1 and Example 1 are shown in Figures 4A to 4C, respectively. Figure 4B shows the results of screening subjects with serum total cholesterol levels greater than 200 mg / dL (i.e., generally considered to have high cholesterol abnormalities), while Figure 4C shows the results of screening subjects with serum total cholesterol levels greater than 200 mg / dL but with LDL-C content. The method for measuring serum total cholesterol and LDL-C levels is the same as that used in "(3) Serum Triglycerides" above, that is, the blood sample is first centrifuged to obtain serum, and then the serum total cholesterol and LDL-C levels are measured using a Cobasproe801 fully automated chemiluminescence immunoassay analyzer.

[0056] As shown in Figure 4A, the serum total cholesterol level of Example 1 on day 28 was significantly lower than that on day 0; conversely, the serum total cholesterol level of Control Example 1 on day 28 was higher than that on day 0. Figure 4B shows that in subjects with high cholesterol, Example 1 demonstrated a better effect in reducing serum total cholesterol levels, and the degree of reduction was also greater than that of Control Example 1. On the other hand, as shown in Figure 4C, in subjects with high cholesterol, the LDL-C level of Example 1 on day 28 was significantly lower than that on day 0, and the degree of reduction was also greater than that of Control Example 1. Therefore, it can be concluded that continuous administration of the *Lactobacillus helveticus* UA881 strain sample of Example 1 for 4 weeks indeed demonstrated an effect in reducing serum total cholesterol and LDL-C levels.

[0057] [(5)] [Pre-meal serum glucose]

[0058] The results of the pre-meal serum glucose levels obtained by the subjects in Control Example 1 and Example 1 are shown in Figure 5 and Table 1 below; Figure 5 and Table 1 below show the results of screening subjects with serum total cholesterol levels greater than 200 mg / dL (i.e., generally considered to have high cholesterol abnormalities). The method for measuring pre-meal serum glucose levels is the same as that used in "(3) serum triglycerides" above, that is, the blood sample is first centrifuged to obtain serum, and then the pre-meal serum glucose levels are measured by a Cobasproe801 fully automated chemiluminescence immunoassay analyzer. Table 1: Pre-meal serum glucose levels measured on day 0 and day 28 of Example 1 and Control Example 1. [Group] [Measurement Time] [Pre-meal serum glucose level] [(mg / dL)] [Compared to the first] [0] [Heaven] [ p ] [value] [average value] [±] [Standard Deviation] Example 1 Day 0 86.57±3.29 -- Day 28 82.14±2.71 0.006509 Compare with Example 1 Day 0 89.00±1.53 -- Day 28 89.86±3.28 0.39

[0059] As shown in Figure 5 and Table 1 above, in subjects with high cholesterol, the pre-meal serum glucose level on day 28 of Example 1 was significantly lower than that on day 0, with a reduction of approximately 5.1%, and the difference was statistically significant (p < 0.01). Conversely, the pre-meal serum glucose level on day 28 of Control Example 1 was higher than that on day 0. This demonstrates that continuous administration of the *Lactobacillus helveticus* UA881 strain sample of Example 1 for 4 weeks did indeed show an effect of reducing pre-meal serum glucose levels, indicating that the *Lactobacillus helveticus* UA881 strain sample of Example 1 did indeed exhibit a blood glucose-lowering effect when administered to obese patients.

[0060] [(6)] [Serum uric acid]

[0061] Figure 6 shows the results of serum uric acid content measurements obtained from subjects in Comparative Example 1 and Example 1. Figure 6 shows the results of screening male subjects with serum uric acid content greater than 7 mg / dL (i.e., generally considered to have abnormal uric acid) and female subjects with serum uric acid content greater than 6 mg / dL (i.e., generally considered to have abnormal uric acid). The method for measuring serum uric acid content is the same as that used in "(3) serum triglycerides" above, that is, the blood sample is first centrifuged to obtain serum, and then the serum uric acid content is measured by a Cobasproe801 fully automated chemiluminescence immunoassay analyzer.

[0062] As shown in Figure 6, in subjects with high uric acid, the serum uric acid level in Example 1 was significantly lower on day 28 than on day 0; conversely, the serum uric acid level in Control Example 1 was higher on day 28 than on day 0. This indicates that continuous administration of the *Lactobacillus helveticus* UA881 strain sample from Example 1 for 4 weeks indeed demonstrated an effect in reducing serum uric acid levels.

[0063] [(7)] [Gut microbiota-related indicators]

[0064] Bacterial DNA was extracted from fecal samples using the QIAamp Fast DNA Stool Mini Kit (purchased from GENOMICS) for analysis. Specifically, following the manufacturer's standard operating procedure, the fecal sample was first centrifuged at 13200×g for 10 minutes, the sample preservation solution was removed, and then the fecal sample was dissolved by shaking with InhibitEX Buffer. Subsequent reagents were added in sequence according to the instruction manual. Finally, the supernatant was washed with a QIAamp spin column and then reconstituted with preheated Elution buffer to obtain the bacterial DNA sample from the feces.

[0065] The library was then constructed and sequenced according to the standard operating procedure of the Illumina 16S Metagenomic sequencing library preparation. First, the V3 and V4 gene fragments of 16S rRNA from intestinal bacteria were amplified by PCR. Then, an index PCR program was performed to attach Illumina adapter and index sequences to both ends of the DNA fragments. Sequencing was then performed using the Illumina Miseq Next Generation Sequencing (NGS) system to obtain the sequencing results. Finally, EzBioCloud software was used to analyze and compare the sequencing results using the 16S rRNA gene sequences of bacteria and archaea integrated by the software, thereby obtaining the results of the microbial composition in the gut. At the same time, OTUs and ACE indicators of the gut microbiota can also be obtained, which can be used to assess the abundance of the gut microbiota.

[0066] On the other hand, the method for detecting fecal butyric acid content is as follows: Take about 0.5 grams of fecal sample and add it to 5 ml of deionized water. Mix well for 2 minutes, then centrifuge at 7000×g and 25°C for 5 minutes. Filter the supernatant through a PTFE membrane with a pore size of 0.45 micrometers. Take 1 ml of the filtrate and add the internal standard isocaproic acid and 100 μL of 50% sulfuric acid solution. Add 1 ml of diethyl ether and mix well for 30 seconds. Centrifuge at 4000×g and 25°C for 2 minutes. Take the supernatant and analyze the butyric acid content using a gas chromatography-mass spectrometry (GC-FID, purchased from Agilent, model 6890A). The gas chromatography conditions are: injection volume of 2 μL, injection temperature of 230°C, column of DB-FFAP (inner diameter: 0.25 mm, length: 30 m), detector temperature of 230°C, and carrier gas of nitrogen.

[0067] [(a) OTUs] [Indicators and] [ACE] [index] [ ]

[0068] The results of OTUs and ACE indices measured in subjects of Comparative Example 1 and Example 1 are shown in Figures 7A and 7B, respectively. Both OTUs and ACE indices represent the abundance of gut microbiota. The vertical axis unit "value" refers to the NGS read count, representing the signal intensity read by NGS. Higher intensity indicates a greater bacterial load; in other words, higher OTUs and ACE indices indicate a richer gut microbiota. It is generally believed that a higher gut microbiota richness contributes to maintaining gut health.

[0069] As can be seen from the results in Figures 7A and 7B, the results of Example 1 on day 28 were significantly higher than those on day 0 for both OTUs and ACE indices. Furthermore, Example 1 also showed a greater degree of improvement in both OTUs and ACE indices than Control Example 1. Therefore, it can be concluded that continuous administration of the Lactobacillus helveticus UA881 strain sample from Example 1 for 4 weeks indeed demonstrated the effect of increasing the abundance of intestinal flora.

[0070] [(b)] [Fecal butyrate]

[0071] The results of measuring fecal butyric acid content in subjects of Control Example 1 and Example 1 are shown in Figure 7C. Butyric acid is a short-chain fatty acid (SCFA), and fecal butyric acid is usually obtained by intestinal microorganisms through the fermentation of dietary fiber. Therefore, fecal butyric acid can be used as an indicator of the metabolic function of beneficial intestinal bacteria. It is generally believed that a higher fecal butyric acid content indicates that it helps maintain intestinal health or alleviate metabolic diseases.

[0072] As can be seen from the 7C results, the butyrate content in Example 1 on day 28 was significantly higher than that on day 0, and the degree to which Example 1 increased the butyrate content was also greater than that in Control Example 1. Therefore, it can be concluded that continuous administration of the *Lactobacillus helveticus* UA881 strain sample from Example 1 for 4 weeks indeed demonstrated the effect of increasing fecal butyrate content to maintain intestinal health.

[0073] [(c)] [Bacteroidetes and] [F / B] [ratio] [ ]

[0074] The results of measuring the number of Bacteroidetes bacteria and the F / B ratio in subjects of Control Example 1 and Example 1 are shown in Figures 7D and 7E, respectively. Bacteroidetes are generally considered beneficial bacteria, capable of producing more short-chain fatty acids by fermenting dietary fiber in the intestines, thus helping to reduce appetite and inhibit fat storage. Therefore, a higher number of Bacteroidetes bacteria helps maintain intestinal health and inhibit obesity. Furthermore, compared to Bacteroidetes (B), Firmicutes (F) bacteria typically constitute a higher proportion in the intestines of obese patients and are considered to promote fat absorption. Therefore, the ratio of Firmicutes bacteria to Bacteroidetes bacteria (F / B ratio) can be used to assess an individual's intestinal condition. A lower F / B ratio indicates that the intestines are less likely to absorb fat, thus helping to prevent obesity and maintain intestinal health.

[0075] As shown in Figure 7D, the number of Bacteroidetes bacteria in Example 1 on day 28 was significantly higher than on day 0, and the degree of increase was also greater than that in Control Example 1. Looking at Figure 7E, the F / B ratio in Example 1 on day 28 was significantly lower than on day 0; conversely, the F / B ratio in Control Example 1 on day 28 was higher than on day 0. Therefore, it can be concluded that continuous administration of the *Lactobacillus helveticus* UA881 strain sample from Example 1 for 4 weeks indeed demonstrated the effects of reducing the risk of obesity and maintaining intestinal health.

[0076] [(d)] [Anaerobic bacteria, Prevotella, and Broutella]

[0077] Figures 7F to 7H show the results of the ratios of the number of anaerobic bacteria (including *Prevotella*, *Bacterium*, and *Brutella*) to the total number of bacteria in subjects of Control Example 1 and Example 1, respectively. The vertical axis unit "ratio" refers to the proportion of the measured specific bacterial genus in the overall gut microbiota. Anaerobic bacteria (including *Prevotella*, *Bacterium*, and *Brutella*) are generally considered probiotics; the higher the proportion of these bacteria in the gut, the more beneficial they are for maintaining gut and metabolic health.

[0078] As shown in Figure 7F, the proportion of anaerobic bacteria in Example 1 on day 28 was significantly higher than on day 0, and the degree of increase was also greater than that in Control Example 1. Looking at Figure 7G, the proportion of Prevotella bacteria in Example 1 on day 28 was significantly higher than on day 0; conversely, the proportion of Prevotella bacteria in Control Example 1 on day 28 was lower than on day 0. Looking at Figure 7H, the proportion of Broutella bacteria in Example 1 on day 28 was significantly higher than on day 0, and the degree of increase was also greater than that in Control Example 1. Therefore, it can be concluded that continuous administration of the *Lactobacillus helveticus* strain UA881 from Example 1 for 4 weeks indeed demonstrated efficacy in maintaining intestinal health.

[0079] In summary, by selecting a specific strain of Lactobacillus helveticus UA881, this invention enables the probiotic composition to have effects such as reducing weight, reducing BMI, reducing waist circumference, reducing visceral fat, reducing fecal lipid content, reducing blood sugar, increasing intestinal flora abundance, increasing fecal butyrate content, increasing the number of Bacteroidetes bacteria in the intestine, reducing the F / B ratio, and / or increasing the number of anaerobic bacteria, Prevotella bacteria, and / or Broutella bacteria in the intestine. In particular, this probiotic composition can be used to prepare hypoglycemic agents, which lower blood sugar in obese patients using the hypoglycemic agents, thus exhibiting a hypoglycemic effect. Therefore, it has the potential to be further developed as a product or treatment for regulating blood sugar and has commercial development value.

[0080] none.

[0081] none.

Claims

1. The use of a probiotic composition in the preparation of a hypoglycemic agent, wherein, The probiotic composition contains a strain of Lactobacillus helveticus UA881, which was obtained from the Licensed Microbial Hosting Center of the Japan Product Evaluation Technology Base, with the accession number NITE BP-03802; the effective dose of the Lactobacillus helveticus UA881 strain in the probiotic composition is 4×10⁹ CFU / day to 6×10⁹ CFU / day, and the Lactobacillus helveticus UA881 strain is in a viable state; The hypoglycemic component was administered to men with a serum total cholesterol level greater than 200 mg / dL, a BMI of 25 kg / m² to 27 kg / m², a pre-meal serum glucose level of 86.57 ± 3.29 mg / dL, and a body fat percentage greater than 25%, or to women with a serum total cholesterol level greater than 200 mg / dL, a BMI of 25 kg / m² to 27 kg / m², a pre-meal serum glucose level of 86.57 ± 3.29 mg / dL, and a body fat percentage greater than 30%.

2. The use as described in claim 1, wherein, The probiotic composition contains metabolites of the Lactobacillus helveticus UA881 strain.

3. The use as described in claim 1, wherein, The blood sugar-lowering components are either food components or pharmaceutical components.

4. The use as described in claim 3, wherein, The pharmaceutical composition also includes a pharmaceutically acceptable carrier.

5. The use as described in claim 4, wherein, This pharmaceutical ingredient is an enteral dosage form.

6. The use as described in claim 5, wherein, This is an oral dosage form that is administered via the intestines.

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