Application of prebiotic component in promoting muscle growth, enhancing muscle strength and improving exercise endurance

Abstract

The invention discloses an application of a prebiotic component in promoting muscle growth, enhancing muscle strength and improving exercise tolerance. The prebiotic component is galactooligosaccharide. Animal experiments show that mice given with galactooligosaccharide show motor function improvement phenomena such as muscle strength increase and fatigue delay in endurance movement in behavioral tests, and the phenomenon that fast muscle and slow muscle fiber components are increased at the same time; the cross-organ integration regulation effect from the intestinal microecology to the whole body physiological state and then to the athletic performance is reflected.

Description

Technical Field

[0001] This invention belongs to the field of sports medicine technology, specifically relating to the application of a prebiotic ingredient in promoting muscle growth, enhancing muscle strength and improving exercise endurance, wherein the prebiotic ingredient is galactooligosaccharide. Background Technology

[0002] Currently, oligosaccharides such as xylooligosaccharide (XOS), galactooligosaccharide (GOS), and fructooligosaccharide (FOS) are known to possess prebiotic properties, regulating gut microbiota and indirectly affecting host metabolism. However, their direct effects on improving motor function have not been systematically studied. Existing research largely focuses on the prebiotic properties of oligosaccharides and their regulatory effects on metabolism or immunity, neglecting their comprehensive impact on motor performance (such as endurance, grip strength, balance, and coordination), and in particular, failing to clarify the differentiated effects of different types of oligosaccharides (such as XOS, GOS, and FOS).

[0003] In the field of sports nutrition research, oligosaccharides, as an important class of functional oligosaccharides, have long been extensively studied due to their prebiotic properties. Oligosaccharides such as XOS, GOS, and FOS can indirectly affect the host's immune function and metabolic health by regulating the balance of gut microbiota. However, traditional research has largely focused on the effects of oligosaccharides on the digestive system, immune regulation, or metabolic diseases, while research on their direct regulation of athletic function is relatively scarce. Currently, the main intervention strategies in sports nutrition include supplementation with branched-chain amino acids and creatine, among other traditional nutrients. While these substances have been shown to enhance muscle synthesis or improve athletic performance, they have limitations such as targeting only a single point of action and the potential for metabolic burden with long-term use.

[0004] In research on the relationship between oligosaccharides and motor function, the most relevant existing technologies mainly involve the indirect influence of oligosaccharides on motor performance through the gut-muscle axis. Existing literature reports that certain oligosaccharides can influence systemic energy metabolism and inflammation levels by regulating the production of short-chain fatty acids by gut microbiota, ultimately positively impacting exercise endurance. However, most of these studies remain at the level of phenomenological observation, lacking systematic comparative assessments of motor function across different types of oligosaccharides, and failing to delve into the specific effects of oligosaccharides on various dimensions of motor performance (including balance, grip, explosive power, and endurance).

[0005] The inventors of this application, through further research, discovered the role of galactooligosaccharides in promoting muscle growth, enhancing muscle strength, and improving exercise endurance, and thus completed this invention. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides an application of prebiotic components in promoting muscle growth, enhancing muscle strength, and improving exercise endurance, and specifically provides the following technical solution: This invention relates to the application of a prebiotic component in enhancing muscle strength and improving exercise endurance, wherein the prebiotic component is galactooligosaccharide.

[0007] In another aspect of the invention, the use of galactooligosaccharides in the preparation of foods and sports drinks that enhance muscle strength and improve exercise endurance is also described.

[0008] Preferably, the application also includes its use in promoting muscle growth.

[0009] Beneficial effects: Animal experiments of this invention show that mice given galactooligosaccharides exhibited improved motor function in behavioral tests, such as increased muscle strength and delayed fatigue during endurance exercise. At the same time, histological examination showed an increase in the content of both fast and slow muscle fibers, which reflects a cross-organ integrated regulatory effect from gut microbiota to systemic physiological state to motor performance. Attached Figure Description

[0010] Figure 1 Molecular weight distribution of galactooligosaccharides; Figure 2 Results of rotarod fatigue test in mice under galactooligosaccharide intervention; Figure 3 Results of gripping test in mice after intervention with galactooligosaccharides; Figure 4 Results of the inverted grid suspension experiment in mice under galactooligosaccharide intervention; Figure 5 Results of instantaneous explosive power test in mice treated with galactooligosaccharide; Figure 6 Results of the constant-speed endurance test in mice under galactooligosaccharide intervention; Figure 7 HE staining results of gastrocnemius muscle in mice treated with galactooligosaccharides; Figure 8 HE staining results of the tibialis anterior muscle in mice under galactooligosaccharide intervention; Figure 9 Masson staining results of gastrocnemius muscle in mice treated with galactooligosaccharides; Figure 10 Masson staining results of the tibialis anterior muscle in mice under galactooligosaccharide intervention Figure 11 Sirius red staining results of gastrocnemius muscle in mice treated with galactooligosaccharides; Figure 12Sirius red staining results of the tibialis anterior muscle in mice under galactooligosaccharide intervention; Figure 13 Results of fast-slow muscle staining in mice treated with galactooligosaccharides; Figure 14 Heatmap of the top 50 most abundant microorganisms at the Genus level; Figure 15 Box plot of abundance of dominant species of Bifidobacterium and Lactobacillus under galactooligosaccharide intervention; Figure 16 Box plots of abundance of several other key bacterial species under galactooligosaccharide intervention. Detailed Implementation

[0011] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0012] Unless otherwise specified, the technical means used in the following embodiments are conventional means well known to those skilled in the art. Unless otherwise specified, the materials, reagents, etc. used in the following embodiments are all commercially available.

[0013] Example 1: 1. Experimental Design and Galacto-oligosaccharide Intervention Program Study subjects: C57BL / 6J mice (8 weeks old, initial weight approximately 22 g) were divided into two groups: NC group (blank control group, administered by gavage with sterile distilled water) and GOS group (galactooligosaccharide intervention group), with 12 mice in each group.

[0014] Intervention method: The GOS group was administered galactooligosaccharide solution (400 mg / kg / d) by gavage daily, with the dosage adjusted according to body weight (e.g., the initial dose was 8.8 mg per mouse).

[0015] Intervention period: 8 weeks, with weight changes recorded concurrently.

[0016] 2. Characterization of galactooligosaccharide structure 2.1 Determination of ash, protein, fat and moisture content in samples The ash, protein, fat and moisture content of galactooligosaccharides were determined according to national standards, and the results are summarized in Table 1 below.

[0017] Table 1. Results of determination of ash, protein, fat and moisture content of xylooligosaccharide The content of each component in the sample meets the quality requirements of GB 1903.27-2022 "National Food Safety Standard for Food Fortifiers - Galacto-oligosaccharides".

[0018] 2.2 Determination of molecular weight and degree of polymerization of samples According to gel permeation chromatography (GPC), also known as size exclusion chromatography (SEC), the molecular weight distribution and degree of polymerization of galactooligosaccharides were determined, and the results are as follows: Figure 1 As shown in Table 2.

[0019] Table 2 Results of determination of molecular weight and degree of polymerization of galactooligosaccharides The results showed that the molecular weight of the galactooligosaccharides in the samples was mainly between 300-1000 g / mol, and the degree of polymerization ranged from 2 to 8. Among them, the content of galactooligosaccharides with a degree of polymerization of 2-5 was relatively high.

[0020] 2.3 Determination of Monosaccharide Composition in Samples The monosaccharide composition of galactooligosaccharides was determined by high-performance liquid chromatography (HPLC). After sample hydrolysis, standard preparation and mixed standard solution preparation, and monosaccharide derivatization, detection was performed under the following chromatographic conditions: T3 column, mobile phase A of acetonitrile, mobile phase B of 0.1 mol / L ammonium acetate solution, detection wavelength of 250 nm, flow rate of 1 ml / min, injection volume of 10 μl, column temperature of 30℃, and external standard method for quantification. Isocratic elution was performed at a ratio of mobile phase A: 19% and mobile phase B: 81%.

[0021] Standard curves were plotted based on the peak areas of each mixed standard sample and their known concentrations. The content of various monosaccharides in galactooligosaccharides was then calculated. The monosaccharides contained only glucose and galactose. The glucose came from the ends of the galactooligosaccharide chains. Rhamnose, glucuronic acid, galacturonic acid, xylose, and arabinose were not detected, which met the requirement that the purity of galactooligosaccharide samples be greater than 70%.

[0022] 3. Motor Function Behavioral Tests 3.1 Rotating bar fatigue test Used to assess the ability of small animals to maintain balance on a rotundi.

[0023] Experimental method: Mice were first warmed up at 5 rpm / min for 5 min, and then accelerated from 5 rpm / min to 40 rpm / min within 300 s. The time (s) when the mice fell from the rotundus was recorded.

[0024] Test results are as follows Figure 2 As shown in the results, the test results indicate that the balance and endurance of the galactooligosaccharide intervention group tended to be enhanced compared with the control group.

[0025] 3.2 Grip Test Assess the animal's gripping / muscle strength level, and evaluate the overall gripping strength of the small animal's forelimbs and hindlimbs.

[0026] Experimental method: Place the mouse on the gripping board and after it has firmly grasped the gripping bar, pull its tail backward until its limbs are completely off the gripping bar, and record the gripping force data (g) at this time.

[0027] Test results are as follows Figure 3 As shown, the test results indicate that the grip strength of the galactooligosaccharide intervention group was significantly improved compared with that of the control group, and the difference between the two groups was significant (p<0.001), indicating that the intervention of galactooligosaccharide can significantly improve the muscle strength of mice.

[0028] 3.3 Inverted Grid Suspension Experiment The ability of animals to grasp and move on an inverted grid is used to assess their upper limb strength and motor coordination.

[0029] Experimental method: The mouse was placed upside down on a 1 cm × 1 cm grid. After it gripped the grid firmly, the time (s) from when it gripped the grid to when it fell was recorded.

[0030] Test results are as follows Figure 4 As shown, the test results indicate significant differences within the groups. Compared with the control group, the fall time in the galactooligosaccharide intervention group tended to increase, suggesting that upper limb strength and coordination were improved under galactooligosaccharide intervention.

[0031] 3.4 Instantaneous Explosive Power Test Using a small animal treadmill with uniform acceleration, the animal's exhaustion speed is tested, and its explosive power is evaluated based on its maximum speed.

[0032] Experimental Method: Mice were acclimatized to the small animal running platform for 3 days at a speed of 5 m / min for 30 min / day, and tested on the 4th day. Before the test, the mice were warmed up at 5 m / min for 5 min, and then the speed was increased to 1 m / min. 2 The acceleration was uniformly increased. When the mouse stood still in the electrical stimulation area (stimulation intensity 0.40 mA) for more than 5 seconds, it was considered to have reached exhaustion, and the maximum movement speed at this time was recorded.

[0033] Test results are as follows Figure 5 As shown, there was no significant difference in maximum speed between the groups, and the average maximum speed of the two groups was quite similar, indicating that galactooligosaccharides have a limited effect on explosive power.

[0034] 3.5 Uniform Speed ​​Endurance Test Animals were allowed to run at a constant speed on a small animal treadmill. The time (s) and distance (m) they ran when they were exhausted were recorded to assess their exercise endurance.

[0035] Experimental Method: After completing the instantaneous explosive power test, rest for one day, and then conduct the test the following day. Warm-up for 5 minutes at 5 m / min, followed by a speed of 3 m / min. 2 Accelerate the vehicle and record the distance and time it takes to reach exhaustion (standard as above).

[0036] Test results are as follows Figure 6 As shown, the test results indicate that the distance of movement in the galactooligosaccharide intervention group was significantly improved compared with that in the control group, and the improvement in motor function in the galactooligosaccharide intervention group was significantly different from that in the control group (p<0.01), indicating that the intervention of galactooligosaccharide can significantly improve the exercise endurance of mice.

[0037] 4. Histopathological analysis 4.1 Muscle Histopathology The results of HE / Masson / Sirius red staining are as follows: Figure 7-12 As shown, the test results indicate that no obvious fibrosis or inflammatory lesions were found, suggesting that galactooligosaccharides did not cause damage to muscle structure.

[0038] 4.2 Staining of fast-twitch muscle components Fast-slow muscle staining results are as follows Figure 13 As shown, the study focused on the soleus muscle (slow-twitch) and the gastrocnemius muscle (fast-twitch). The results showed that the expression levels of fiber components in both slow-twitch and fast-twitch muscles were significantly increased, suggesting that galacto-oligosaccharides may improve athletic performance by promoting muscle growth.

[0039] 5. Metagenomic Mechanism Analysis: Key Microbial Communities and Functional Associations 5.1 Qualitative identification of key bacterial genera using heatmaps After 8 weeks of galactooligosaccharide intervention experiment, mouse fecal samples were collected for metagenomic sequencing (12 mice were sampled in each group, 3 mice / mixed sample, i.e. 4 samples / group, for a total of 8 samples, control group mice (CON1-4) and galactooligosaccharide intervention group mice (XOS1-4).

[0040] Test results are as follows Figure 14 As shown, the test results indicate that *Sangerella* spp. ( Sangeribacter ) 、 Clostridium rumenans ( Ruminiclostridium ) 、 Rosbyella spp. Roseburia ) 、 Eubacterium ( Eubacterium ) 、 Hengate ( spp.) Hungatella ), and two probiotic genera, Bifidobacterium ( Bifidobacterium ) and Lactobacillus genus ( LactobacillusThe abundance of galactooligosaccharides (GOS) showed significant changes, providing insights into the identification of key bacterial species regulated by GOS.

[0041] 5.2 Probiotic Abundance Analysis Test results are as follows Figure 15 As shown, the test results indicate that Bifidobacterium spp. ( Bifidobacterium Two representative bacterial species Bifidobacterium pseudolongum and Bifidobacterium sp. The abundance of *Lactobacillus* in the experimental groups was lower than that in the control groups, and there were significant differences between the groups. Lactobacillus Two representative bacterial species Lactobacillus johnsonii and Lactobacillus intestinalis The abundance in the experimental group was also lower than that in the control group, among which... Lactobacillus intestinalis The abundance in the experimental group was significantly different from that in the control group (p < 0.01).

[0042] 5.3 Other Key Microbial Communities Test results are as follows Figure 16 As shown, the test results indicate that under the intervention of galactooligosaccharides, Rosbyraceae ( Roseburia ) 、 Hengate ( spp.) Hungatella The abundance of the most abundant bacterial species in the experimental group was lower than that in the control group, while the abundance of *Sangerella* spp. was lower. Sangeribacter ) 、 Clostridium rumenans ( Ruminiclostridium The abundance of representative bacterial species of galactooligosaccharides (GOS) increased. This indicates that GOS alters the structure and competitive relationships of the gut microbiota. When GOS enters the gut, it is efficiently metabolized by specific bacterial groups to produce short-chain fatty acids (SCFAs) and lower the local pH value, creating an ecological environment favorable to both "GOS users" and "fatty acid producers," while inhibiting "non-GOS users." Ultimately, this results in an increase in the abundance of the former and a decrease in the abundance of the latter. 6. Conclusion and Analysis 6.1 The effect of galactooligosaccharides on improving motor function: Galacto-oligosaccharides can effectively improve the motor functions of mice, such as muscle strength, upper limb coordination, and exercise endurance, and significantly promote muscle growth in mice.

[0043] 6.2 Bacterial-muscle axis mechanism hypothesis: The improvement in motor abilities such as muscle strength, upper limb coordination, and exercise endurance in mice after intervention with galactooligosaccharides (GOS), as well as the increase in fast and slow muscle fiber content, is essentially a systemic optimization achieved by reshaping the gut microbiota and activating the "gut microbiota-muscle axis." GOS, as a prebiotic, is efficiently fermented by specific short-chain fatty acid-producing bacteria, producing metabolites such as butyric acid, propionic acid, and acetic acid. These substances not only provide energy for intestinal epithelial cells and repair the barrier to reduce chronic inflammation caused by endotoxins entering the bloodstream, but also enter the body through blood circulation, improving energy homeostasis, enhancing mitochondrial function, and reducing oxidative stress, thereby creating a more favorable metabolic environment for muscles. Simultaneously, microbiota-derived neurotransmitter precursors and metabolic signals regulate central motivation and neuromuscular coordination through the gut microbiota-brain axis. Under these combined effects, mice exhibited improved motor function in behavioral tests, including enhanced muscle strength and upper limb coordination, and delayed fatigue during endurance exercise, along with increased muscle fiber content. This reflects a cross-organ integrated regulatory effect from gut microbiota to systemic physiological state and then to motor performance.

[0044] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. The application of a prebiotic ingredient in enhancing muscle strength and improving upper limb coordination, wherein the prebiotic ingredient is galactooligosaccharide.

2. The application according to claim 1, wherein the application further includes promoting muscle growth.

3. The application of a type of galactooligosaccharide in the preparation of foods and sports drinks that enhance muscle strength and improve upper limb coordination.

4. The application according to claim 3, wherein the food and sports drink are used to promote muscle growth.

Images