Diglyceride composition and application thereof

Abstract

The invention relates to a functional nutritional composition, in particular to an anti-fatigue diglyceride composition, which is characterized by comprising (1) diglyceride, (2) isomaltooligosacharide, (3) sodium caseinate, (4) soybean phospholipid, (5) sodium citrate, (6) potassium citrate, (7) calcium ascorbate, (8) silicon dioxide, (9) mogroside, (10) vitamin B1 and (11) vitamin B2. The composition disclosed by the invention can be used as an anti-fatigue beverage, a nutritional health-care product, a food for special diets, a food for special medical purposes and the like.

Description

Technical Field

[0001] This invention belongs to the field of food / health products and relates to a functional nutritional composition, more specifically, to a diglyceride composition with anti-fatigue effects. Background Technology

[0002] Fatigue is a ubiquitous physiological phenomenon and a complex physiological and biochemical process. Fatigue can be divided into two types: central fatigue and peripheral fatigue (McKenna, MJ and M. Hargreaves, Resolving fatigue mechanisms determining exercise performance: integrative physiology at its finest! J ApplPhysiol (1985), 2008. 104(1): p. 286-7.). Central fatigue is caused by a disorder of the motor nerve center, resulting in a decrease in the excitability of the motor nerves; peripheral fatigue manifests as muscle fatigue, i.e., a decrease in muscle strength (Zając, A., et al., Central and Peripheral Fatigue During Resistance Exercise - ACritical Review. J Hum Kinet, 2015.49: p. 159-69.).

[0003] Diacylglycerol (DAG) is a product obtained by esterifying two fatty acid molecules and one glycerol molecule. As a functional oil, it has been recognized as a GRAS (generally recognized as safe) food ingredient. As a recognized safe oil with no side effects, the scientific research and food applications of diglycerol will receive increasing attention.

[0004] Currently, the market offers a wide variety of products for relieving fatigue, including chemical drugs, health supplements, and traditional Chinese medicine preparations. While chemical drugs such as caffeine can provide short-term alertness and fatigue relief, long-term use may lead to side effects such as dependence and tolerance. The health supplement market is somewhat chaotic, with the efficacy and safety of some products lacking sufficient scientific validation. In particular, products that simultaneously relieve both central and peripheral fatigue are rare. Therefore, there is an urgent need to provide a highly effective anti-fatigue composition. Summary of the Invention

[0005] The purpose of this invention is to provide an anti-fatigue composition. In particular, this invention creatively selects to use diglycerides in combination with other ingredients, which greatly enhances the anti-fatigue function of the composition.

[0006] The present invention provides a diglyceride composition comprising the following components: diglyceride, isomaltooligosaccharide, sodium caseinate, soybean lecithin, sodium citrate, potassium citrate, calcium ascorbate, silicon dioxide, mogroside, vitamin B1, and vitamin B2.

[0007] Furthermore, the diglyceride composition, by weight, comprises the following components: 35-65 parts diglyceride, 35-55 parts isomaltooligosaccharide, 1.5-4.5 parts sodium caseinate, 0.05-1 part soybean lecithin, 0.05-1 part sodium citrate, 0.05-1 part potassium citrate, 0.05-0.5 parts calcium ascorbate, 0.1-0.5 parts silicon dioxide, 0.05-0.2 parts mogroside, 0.001-0.01 parts vitamin B1, and 0.001-0.01 parts vitamin B2.

[0008] Furthermore, the diglyceride composition, by weight, comprises the following components: 50 parts diglyceride, 44.8 parts isomaltooligosaccharide, 2.8 parts sodium caseinate, 0.5 parts soybean lecithin, 0.6 parts sodium citrate, 0.5 parts potassium citrate, 0.3 parts calcium ascorbate, 0.3 parts silicon dioxide, 0.1 parts mogroside, 0.05 parts vitamin B1, and 0.05 parts vitamin B2.

[0009] Furthermore, the diglyceride composition can be prepared as an oral formulation.

[0010] Furthermore, oral preparations include various dosage forms, such as powders, ointments, granules, tablets, pills, or capsules.

[0011] The present invention also provides the application of a diglyceride composition in beverages, nutritional supplements, special dietary foods, and foods for special medical purposes.

[0012] Furthermore, the products derived from this application are used to alleviate peripheral fatigue.

[0013] Furthermore, the products derived from this application are used to relieve central nervous system fatigue.

[0014] Furthermore, it can alleviate peripheral fatigue by increasing muscle mass, improving exercise endurance, reducing BUN content, increasing LG and MG content, reducing MDA content, enhancing SOD activity, and relieving gastrocnemius muscle damage.

[0015] Furthermore, it can alleviate central nervous system fatigue by reducing the level of the neurotransmitter γ-aminobutyric acid (GABA), increasing the content of glutamate (Glu), increasing the number of grids crossed in the mining experiment, and improving the damage of hippocampal neurons.

[0016] The diglyceride composition provided by this invention can relieve both peripheral and central nervous system fatigue; and while exerting its anti-fatigue effect, it has no significant adverse effects on the body's normal physiological functions and growth and development, has high safety, and the formulation and preparation method are relatively simple. Attached Figure Description

[0017] Figure 1 The effect of the composition on body weight (A) and weight gain (B) in fatigued mice.

[0018] Figure 2 The effect of the composition on the exhaustive swimming time of a fatigue model mouse.

[0019] Figure 3 The effect of the composition on blood urea nitrogen (BUN) levels in fatigue model mice was investigated.

[0020] Figure 4 The effect of the composition on liver glycogen (A) and muscle glycogen (B) in fatigue model mice.

[0021] Figure 5 The effect of the composition on superoxide dismutase (SOD) and malondialdehyde (MDA) in fatigue model mice.

[0022] Figure 6 The effect of the composition on the gastrocnemius muscle structure (HE staining (×400)) of fatigue model mice.

[0023] Figure 7 The effects of the composition on the neurotransmitters γ-aminobutyric acid (GABA) (A) and glutamate (Glu) (B) in fatigue model mice.

[0024] Figure 8 The effect of the composition on the fatigue model mouse mine field experiment (number of grid crossings and number of cheek scratches).

[0025] Figure 9 The effect of the composition on the hippocampus of fatigue model mice (HE staining (×200)). Detailed Implementation

[0026] The present invention will be further described below with reference to specific examples and accompanying drawings. Those skilled in the art should understand that the examples and accompanying drawings are only for a better understanding of the present invention and are not intended to make any limitations.

[0027] Example 1: The composition of the composition is as follows: 50 parts diglyceride, 44.8 parts isomaltooligosaccharide, 2.8 parts sodium caseinate, 0.5 parts soybean lecithin, 0.6 parts sodium citrate, 0.5 parts potassium citrate, 0.3 parts calcium ascorbate, 0.3 parts silicon dioxide, 0.1 parts mogroside, 0.05 parts vitamin B1 and 0.05 parts vitamin B2. In preparation, 50 parts of diglycerides and 0.5 parts of soybean lecithin were mixed and heated to 45°C, and stirred until completely dissolved. Then, 44.8 parts of isomaltooligosaccharide and 2.8 parts of sodium caseinate were added, and the mixture was stirred at 300 r / min for 15 min until a uniform suspension was formed. Then, 0.6 parts of sodium citrate, 0.5 parts of potassium citrate, 0.3 parts of calcium ascorbate, and 0.3 parts of silicon dioxide were added sequentially, and the mixture was stirred for another 10 min. Finally, 0.1 parts of mogroside, 0.05 parts of vitamin B1, and 0.05 parts of vitamin B2 were added, and the mixture was stirred for 5 min and then cooled to room temperature to form a powder.

[0028] Example 2: The composition of the composition is as follows: 35 parts diglyceride, 55 parts isomaltooligosaccharide, 1.5 parts sodium caseinate, 0.05 parts soybean lecithin, 0.05 parts sodium citrate, 0.05 parts potassium citrate, 0.05 parts calcium ascorbate, 0.1 parts silicon dioxide, 0.05 parts mogroside, 0.001 parts vitamin B1 and 0.001 parts vitamin B2. In preparation, 35 parts of diglycerides and 0.05 parts of soybean lecithin were mixed and heated to 45°C, and stirred until completely dissolved. Then, 55 parts of isomaltooligosaccharide and 1.5 parts of sodium caseinate were added, and the mixture was stirred at 300 rpm for 15 minutes to form a uniform suspension. Then, 0.05 parts of sodium citrate, 0.05 parts of potassium citrate, 0.05 parts of calcium ascorbate, and 0.1 parts of silicon dioxide were added sequentially, and the mixture was stirred for another 10 minutes. Finally, 0.05 parts of mogroside, 0.001 parts of vitamin B1, and 0.001 parts of vitamin B2 were added, and the mixture was stirred for 5 minutes and then cooled to room temperature to form a powder.

[0029] Example 3: Composition of the composition: 65 parts of diglyceride, 35 parts of isomaltooligosaccharide, 4.5 parts of sodium caseinate, 1 part of soy lecithin, 1 part of sodium citrate, 1 part of potassium citrate, 0.5 part of calcium ascorbate, 0.5 part of silicon dioxide, 0.2 part of mogroside, 0.01 part of vitamin B1 and 0.01 part of vitamin B2. During preparation, 65 parts of diglyceride and 1 part of soy lecithin are mixed and heated to 45°C, stirred until completely dissolved, 35 parts of isomaltooligosaccharide and 4.5 parts of sodium caseinate are added, and stirred at a speed of 300 r / min for 15 min until a uniform suspension is formed. Then, 1 part of sodium citrate, 1 part of potassium citrate, 0.5 part of calcium ascorbate, and 0.5 part of silicon dioxide are added in sequence, and stirring is continued for 10 min. Finally, 0.2 part of mogroside, 0.01 part of vitamin B1, and 0.01 part of vitamin B2 are added, stirred for 5 min, and then cooled to room temperature to prepare a powder.

[0030] Verification of anti-fatigue effect: The composition prepared in Example 1 is taken below for verification of anti-fatigue effect.

[0031] Experimental animals: 50 male ICR mice of SPF grade, 6 - 8 weeks old, with animal license number SCXK(Yue) 2020 - 0051, weighing 18 - 22 g. They are adaptively fed for 7 d, during which they have normal diet and water, with a light cycle of 12 / 12 h (day / night), the room temperature is controlled at 20°C - 26°C, and the humidity is controlled in the range of 40% - 70%, and the animal room is kept clean and hygienic.

[0032] Establishment of fatigue model: The chronic multi-factor composite stress stimulation method is used to establish a fatigue model (chronic restraint + chronic unpredictable mild stress + excessive exercise), simulating pathogenic factors such as modern human physical overloading, instinct frustration, and ultra-high-pressure mental state, to replicate the irregular negative factors in all aspects of humans to the greatest extent and induce fatigue symptoms, mainly including: chronic restraint for 5 h, fasting and water deprivation for 24 h, wet bedding for 24 h, forced swimming for 10 min, running on a rotating wheel for 15 min, shaking the cage for 5 min, crowding for 24 h, and circadian rhythm inversion. Two stimuli are randomly given every day, and the same stimulus does not appear for two consecutive days. The model is established for 28 days in total.

[0033] The grouping and drug administration of the mice are shown in Table 1 below: Table 1 Grouping and drug administration of mice

[0034] Weighted swimming test: This test simulates high-intensity exercise and assesses exercise endurance and fatigue levels, making it an effective tool for studying exercise-induced fatigue. A mouse is placed in a heated bath with a lead weight approximately 5% of its body weight wrapped around its tail. The mouse swims in the bath (water temperature 25 ± 1°C, water depth ≥ 30 cm, water surface constantly stirred to ensure the mouse remains in a swimming state). Timing begins, and the mouse is considered to have reached exhaustion when its head is submerged for 10 seconds. This is the mouse's exhaustion swimming time.

[0035] Mine Field Experiment: The mine field experiment assesses the degree of central nervous system fatigue in mice by recording their spontaneous activities within an open area. The experimental method is a slight modification of the de Oliveira method. Mice are placed in a black box with 25 squares (50cm × 50cm × 45cm), and their movements are recorded over 5 minutes.

[0036] Weight measurement: The weight of each mouse was measured and recorded using a balance at 0, 7, 14, 21 and 28 days.

[0037] Determination of organ index in mice: After removing each organ (whole brain tissue, heart, liver, kidney), they were cleaned with physiological saline, dried, and weighed.

[0038] Organ index (%) = Determination of m1 / m0 number In the formula, m1 represents the weight of each organ of the mouse (g), and m0 represents the body weight of the mouse (g).

[0039] Determination of mouse muscle tissue index: After dissecting the mouse muscle tissue, clean it with physiological saline, dry it, and weigh it.

[0040] Muscle tissue index (%) = m1 / m0 × 100 In the formula, m1 represents the weight of the mouse's left and right legs (g), and m0 represents the mouse's body weight (g).

[0041] ELISA detection and biochemical index determination: After the last administration of the drug to establish the mouse model, blood was collected from mice and tissues were dissected. ELISA and biochemical index determination were then performed. Serum BUN levels and liver and muscle glycogen levels were detected using biochemical kits. ELISA kits were used to detect Glu and GABA levels in brain tissue and MDA and SOD enzyme activities in liver tissue.

[0042] Histopathological HE staining sections: After dissection, hippocampal and muscle tissues from the mouse brain were obtained and stained using a hematoxylin-eosin (HE) staining kit. The results were observed under an optical microscope.

[0043] Data Processing: All experiments were repeated at least three times, and a p-value < 0.05 was considered statistically significant. Graphpad Prism 10 software (Graphpad Software, USA) was used for data analysis and plotting. T-tests were used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. Significance comparison between the model group and the control group: "#" indicates p < 0.05; "##" indicates p < 0.01; "###" indicates p < 0.001; "####" indicates p < 0.0001. Significance comparison between the dosage group and the model group: "Indicates p < 0.05;" "Indicates p < 0.01;" "Indicates p < 0.001;" " indicates p < 0.0001.

[0044] Mouse body weight results: Figure 1 It was found that the average weight of mice before drug administration was 32.06 ± 2.51 g. After drug administration began, the weight of mice in all five groups steadily increased. After drug administration ended, the weights of mice in the blank group, model group, Rhodiola rosea positive group, low-dose combination group, and high-dose combination group were 41.82 ± 3.44 g, 38.59 ± 3.67 g, 37.99 ± 1.98 g, 42.23 ± 2.04 g, and 40.18 ± 2.02 g, respectively. The weight gains of mice in each group were 9.56 ± 2.26 g, 6.01 ± 1.25 g, 6.77 ± 1.68 g, 8.79 ± 1.18 g, and 8.76 ± 1.45 g, respectively. Compared with day 0, the mice in the model group had the slowest weight gain (p<0.01), indicating that the fatigue model was successfully established. The weight gain of the mice in the composition administration group was significantly higher than that in the model group (p<0.05), but similar to the blank group (normal mouse group), indicating that the composition can improve the weight loss of fatigued mice and restore them to normal levels.

[0045] Results of organ index experiments in mice: As shown in Table 2, there were no significant differences in organ indices between the model group and the control group, indicating that fatigue had little effect on the organs of mice. Compared with the model group, there were no significant differences in organ indices between the Rhodiola rosea positive group and the combined drug administration group, indicating that drug intervention also had no significant effect on the organs of fatigued mice.

[0046] Table 2. Organ Index of Mice (%)

[0047] Effect of the composition on relieving peripheral fatigue Mouse muscle tissue index (Table 4): Compared with the blank group, the model group mice showed a significant decrease in muscle tissue (p<0.05), indicating successful model establishment and reduced muscle mass in fatigued mice. Compared with the model group, the Rhodiola rosea positive group and the combination drug administration group showed a significant increase in muscle tissue (p<0.05), suggesting that the combination can promote muscle growth and improve motor ability.

[0048] Table 4. Mouse muscle tissue index (%)

[0049] Mouse exhaustive swimming time: The exhaustive swimming test is commonly used to evaluate the anti-fatigue activity of compounds; the longer the time, the more significant the anti-fatigue effect of the compound. Figure 2 Before drug administration, there was no statistically significant difference in the exhaustive swimming time among the groups (p>0.05), with an average of 7.31 ± 2.12 min. Twenty-eight days after drug administration, the exhaustive swimming times in each group were 7.94 ± 0.99 min, 4.16 ± 1.7 min, 6.26 ± 2.46 min, 7.13 ± 3.07 min, and 8.2 ± 3.36 min, respectively. Compared with the control group, the exhaustive swimming time was reduced in the model group, and significantly increased in the positive control group and the high-dose group of the composition (p<0.01, p<0.05, p<0.05). These results indicate that the fatigue mouse model in this experiment is reliable, and the diglyceride composition has a similar effect to Rhodiola rosea, both significantly improving fatigue and enhancing swimming endurance in fatigued mice, exhibiting a certain dose-dependent characteristic. This may promote muscle growth and improve athletic ability to some extent, thereby improving the athletic performance of mice.

[0050] Blood urea nitrogen (BUN) levels in mice after exercise: BUN is an important indicator of fatigue. During high-intensity exercise, muscle oxygen consumption increases, and the body switches to anaerobic metabolism due to hypoxia. Glycolysis accelerates, leading to a large accumulation of lactic acid, which lowers the pH of muscle tissue and blood, affecting cardiovascular and skeletal muscle function, and is a significant cause of fatigue. Lactate dehydrogenase can reflect lactic acid levels, indirectly reflecting exercise status. During high-intensity exercise, if the body does not replenish glucose in time, it will consume protein for energy. Urea is the final product of protein metabolism; therefore, blood urea levels increase significantly during strenuous exercise. The more severe the fatigue, the higher the blood urea nitrogen concentration. Figure 3As shown, 28 days after drug administration, the BUN content in the model group mice was significantly increased compared with the control group (p<0.0001). This indicates that the fatigue mouse model was successful. Compared with the control group, the BUN content in mice increased after intervention with Rhodiola rosea and the combination; however, compared with the model group, the BUN content in mice was significantly reduced (p<0.01, p<0.05, p<0.01). These results indicate that the multi-factor induction method for constructing a fatigue mouse model is reliable, and the combination has a similar effect to Rhodiola rosea, partially reversing the increased BUN content in mice due to fatigue.

[0051] Liver glycogen (LG) and muscle glycogen (MG) in mice after exercise: Glycogen is an important energy source during exercise. When blood glucose levels decrease, liver glycogen is converted into glucose, while muscle glycogen directly supplies energy to muscle tissue via anaerobic glycolysis. Therefore, glycogen content is an important indicator for assessing fatigue capacity. Figure 4 As shown, 28 days after drug administration, compared with the control group, the liver glycogen and muscle glycogen contents of mice in the model group were significantly decreased (p<0.01, p<0.01). This indicates that the fatigue mouse model was successful. However, compared with the model group, after intervention with high doses of Rhodiola rosea and the combination, the liver glycogen and muscle glycogen contents of mice significantly recovered (p<0.05, p<0.05, p<0.001, p<0.05). These results indicate that the fatigue mouse model is reliable, and the effects of the combination are comparable to those of Rhodiola rosea.

[0052] Superoxide dismutase (SOD) and malondialdehyde (MDA) in mouse muscles after exercise: After strenuous exercise, the level of reactive oxygen species in the body increases, leading to intensified lipid peroxidation in the liver and muscle tissue. MDA is one of the metabolic products of lipid peroxidation during cell membrane degradation. Therefore, MDA is an important indicator of the level of oxidative stress in an organism and can indirectly reflect the degree of oxidative damage to lipids on the cell membrane. Higher MDA levels indicate reduced antioxidant capacity, decreased ability to scavenge free radicals, and more severe lipid peroxide fatigue. Figure 5 As shown, 28 days after drug administration, the MDA level in the model group mice was significantly increased compared with the blank group (p<0.01), indicating that the fatigue mouse model was successful. However, compared with the model group, the MDA level in mice significantly decreased after intervention with Rhodiola rosea and its combination (p<0.01, p<0.05, p<0.05).

[0053] On the other hand, the presence of reactive oxygen species (ROS) in the body leads to protein oxidation. Under normal circumstances, ROS are effectively eliminated by the antioxidant defense system. However, when the oxidative and antioxidant systems are out of balance, ROS-induced oxidative stress damages biomolecules in the body, leading to DNA damage and fatigue. Superoxide dismutase (SOD) can catalyze the production of H2O2 and O2 from ROS, and glutathione peroxidase (GSH-px) can further catalyze the production of H2O2 and O2, protecting body tissues from free radical attacks. Therefore, enhancing the activity of these antioxidant enzymes can help alleviate fatigue. Figure 5 As shown, 28 days after drug administration, the SOD level in the model group mice was significantly lower than that in the control group (p<0.0001). This indicates that the multi-factor induction method for constructing a fatigue mouse model was successful. Compared with the model group, the SOD level in mice significantly increased after intervention with Rhodiola rosea and the combination (p<0.0001, p<0.01, p<0.01). These results indicate that the fatigue mouse model is reliable, and the combination has a similar effect to Rhodiola rosea, which can alleviate fatigue by increasing the activity of antioxidant enzymes in the body.

[0054] Muscle structure in mice after exercise: The gastrocnemius muscle of the hind leg of mice was dissected, and histological changes in the gastrocnemius muscle were observed by H&E staining to reflect the mouse's motor ability. Figure 6 As shown, compared with the control group, many skeletal muscle cells in the model group mice were loosely arranged, with ruptured nuclei, numerous intracellular cracks, and severe necrosis. In the Rhodiola rosea-positive group, local muscle fibers showed cracks. In the low-dose administration group, skeletal muscle cells were loosely arranged, with many cells showing slight swelling and some cracks. In contrast, the gastrocnemius muscle cells in the high-dose administration group were tightly arranged, with only slight cracks. Therefore, Rhodiola rosea and its combination can protect the gastrocnemius muscle of mice to a certain extent, relieve fatigue, and enhance motor ability.

[0055] In summary, the diglyceride composition can alleviate peripheral fatigue in fatigued mice by increasing muscle mass, improving exercise endurance, reducing BUN content, increasing LG and MG content, reducing MDA content, enhancing SOD activity, and relieving gastrocnemius muscle damage.

[0056] Effect of the composition on relieving central nervous system fatigue Neurotransmitters in the mouse brain: Glutamate (Glu), as a precursor to the central inhibitory neurotransmitter gamma-aminobutyric acid (GABA), is synthesized into GABA during amino acid metabolism in vivo under the action of glutamate decarboxylase. GABA is released at synaptic terminals, acting on the cell bodies of postsynaptic neurons, inhibiting neuronal excitatory activity, inducing inhibitory processes in the brain, and ultimately leading to central fatigue. As shown in Figure 7, after 28 days of treatment, compared with the control group, the GABA content in the model group mice was significantly increased (p<0.0001), and the Glu content rapidly decreased (p<0.001). This indicates that the multi-factor induction method for constructing a fatigue mouse model was successful. Compared with the model group, after intervention with Rhodiola rosea and its combination, the GABA content in mice was significantly decreased (p<0.0001, p<0.05, p<0.01), and the Glu content was significantly increased (p<0.01, p<0.05). The above results indicate that the fatigue mouse model constructed in this experiment is reliable, and the composition has a similar effect to Rhodiola rosea, and can reverse the changes in neurotransmitters γ-aminobutyric acid (GABA) and glutamate (Glu) caused by fatigue.

[0057] In the mouse mine experiment, a decrease in the number of times an animal traverses a grid typically indicates a decline in its activity level, possibly due to weakened motor coordination and motivation caused by central nervous system fatigue. An increase in cheek-grabbing can reflect anxiety or discomfort behaviors due to fatigue; central nervous system fatigue affects mood regulation, leading to an increase in similar behaviors. These behavioral changes are closely related to central nervous system fatigue, which affects motor function, mood, and exploratory abilities, thereby altering the performance of these behaviors. Therefore, observing these indicators can indirectly assess the degree of central nervous system fatigue. Figure 8 Before modeling and drug administration, there were no statistically significant differences in the number of squares crossed and the number of times mice scratched their cheeks among the different groups (p > 0.05). Twenty-eight days after modeling and drug administration, the number of squares crossed by the five groups were 186 ± 74, 134 ± 29, 181 ± 63, 154 ± 51, and 176 ± 56, respectively. Compared with the control group, the number of squares crossed by the model group was significantly reduced (p < 0.01), indicating successful modeling and a significant slowdown in the mice's autonomous exploration behavior. Furthermore, compared with the model group, the number of squares crossed by the positive control group and the high-dose group of the composition was significantly increased (p < 0.05), indicating that the composition had an intervention effect on the fatigue behavior of fatigued mice, which resulted in a reduced number of squares crossed. Regarding the number of times mice scratched their cheeks, there was no significant difference among the groups after 28 days of modeling and drug administration (p > 0.05), indicating that the composition had little effect on the number of times mice scratched their cheeks.

[0058] Mouse hippocampal tissue: Figure 9The images show the HE staining results of coronal sections of mouse hippocampal tissue under a microscope at 200x magnification. In the control group, hippocampal neurons in the brain tissue were neatly arranged, with clear, dense, and uniform size, and clearly defined nuclei. In the model group, hippocampal neurons in the brain tissue were loosely and disordered, with increased gaps between cells, swollen cytoplasm, and deeper cytoplasmic staining. Cells were deformed and atrophied, with significant loss of granule cells and numerous scattered inflammatory cells, indicating successful modeling and inducing hippocampal tissue damage. Compared to the model group, intervention with Rhodiola rosea and the combined drug significantly improved hippocampal neuronal cell activity in mouse brain tissue. Although the neuronal arrangement was looser than in the control group, there was less deeper cytoplasmic staining and less granule loss, and the number of inflammatory cells was reduced. These results indicate that the multi-factor-induced fatigue mouse model used in this experiment is reliable, and the combined drug has comparable effects to Rhodiola rosea, partially improving fatigue-induced hippocampal tissue damage.

[0059] In summary, the diglyceride composition can alleviate central nervous system fatigue by reducing the level of the neurotransmitter γ-aminobutyric acid (GABA), increasing the content of glutamate (Glu), increasing the number of grids crossed in the mine field experiment, and improving the damage of hippocampal neurons.

[0060] In this way, the diglyceride composition can relieve both peripheral and central nervous system fatigue; and while exerting its anti-fatigue effect, it has no significant adverse effects on the body's normal physiological functions and growth and development, with high safety, and the formulation and preparation method are relatively simple.

[0061] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A diglyceride composition, characterized in that, It includes the following components: diglycerides, isomaltooligosaccharides, sodium caseinate, soybean lecithin, sodium citrate, potassium citrate, calcium ascorbate, silicon dioxide, mogrosides, vitamin B1, and vitamin B2.

2. The diglyceride composition according to claim 1, characterized in that, By weight, it consists of the following components: 35-65 parts diglycerides, 35-55 parts isomaltooligosaccharide, 1.5-4.5 parts sodium caseinate, 0.05-1 part soybean lecithin, 0.05-1 part sodium citrate, 0.05-1 part potassium citrate, 0.05-0.5 parts calcium ascorbate, 0.1-0.5 parts silicon dioxide, 0.05-0.2 parts mogroside, 0.001-0.01 parts vitamin B1 and 0.001-0.01 parts vitamin B2.

3. The diglyceride composition according to claim 1, characterized in that, By weight, it consists of the following components: 50 parts diglyceride, 44.8 parts isomaltooligosaccharide, 2.8 parts sodium caseinate, 0.5 parts soybean lecithin, 0.6 parts sodium citrate, 0.5 parts potassium citrate, 0.3 parts calcium ascorbate, 0.3 parts silicon dioxide, 0.1 parts mogroside, 0.05 parts vitamin B1, and 0.05 parts vitamin B2.

4. The diglyceride composition according to any one of claims 1-3, characterized in that, The diglyceride composition can be prepared as an oral formulation.

5. The oral preparation according to claim 4 includes powder, ointment, granules, tablets, pills or capsules.

6. The use of the diglyceride composition according to any one of claims 1-3 in beverages, nutritional supplements, special dietary foods, foods for special medical purposes, etc.

7. The application according to claim 6, characterized in that, The product obtained from this application is used to alleviate peripheral fatigue.

8. The application according to claim 6, characterized in that, The product obtained from this application is used to relieve central nervous system fatigue.

9. The application according to claim 7, characterized in that, The relief of peripheral fatigue is achieved by increasing muscle mass, improving exercise endurance, reducing BUN content, increasing LG and MG content, reducing MDA content, enhancing SOD activity, and alleviating gastrocnemius muscle damage.

10. The application according to claim 8, characterized in that, The relief of central nervous system fatigue is achieved by reducing the level of the neurotransmitter γ-aminobutyric acid (GABA), increasing the content of glutamate (Glu), increasing the number of grids crossed in the mining experiment, and improving the damage of hippocampal neurons.

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