Application of composition compounded with lycium ruthenicum probiotics in preparation of anti-inflammatory, taurine metabolism promoting and anti-fatigue products

Abstract

The invention belongs to the technical field of anti-inflammation and anti-fatigue, and particularly relates to application of a composition compounded with lycium ruthenicum probiotics to preparation of products capable of resisting inflammation, promoting taurine metabolism and resisting fatigue. The lycium ruthenicum and lycium ruthenicum probiotic compound composition not only can synergistically reduce the release of proinflammatory factors IL-6, TNF-alpha and IL-1beta, scavenge free radicals and improve the oxidation resistance, but also can ferment and metabolize various components such as organic acid in lycium ruthenicum freeze-dried powder, promote taurine synthesis, improve the immunity of the lycium ruthenicum and the lycium ruthenicum probiotics, and improve the immunity of the lycium ruthenicum. Therefore, the anti-oxidation and anti-inflammatory effects are further exerted, the obvious anti-fatigue effect is synergistically exerted, and good application prospects are achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of anti-inflammatory and anti-fatigue, and specifically relates to an application of a composite black wolfberry probiotic composition in the preparation of anti-inflammatory, taurine metabolism-promoting and anti-fatigue products. Background Art

[0002] Chronic inflammation is a long-term, low-grade, systemic inflammatory state caused by the body's response to persistent damaging factors or abnormal immune responses. Unlike acute inflammation, it is characterized by monocyte / macrophage infiltration, tissue fibrosis, and impaired repair, and is a common pathological basis for many chronic diseases. Elevated levels of cytokines such as IL-1β and IL-6 suppress activity in the brain's frontal cortex, increasing the body's perception of fatigue.

[0003] While traditional anti-inflammatory drugs (such as nonsteroidal anti-inflammatory drugs) can quickly relieve symptoms, long-term use can easily lead to gastrointestinal damage and drug resistance. Taurine, an endogenous antioxidant and energy metabolism regulator, is susceptible to intestinal flora imbalance and enterohepatic circulation disorders, and its synthesis is susceptible to problems such as low bioavailability and a single metabolic regulation target. Furthermore, anti-fatigue products often rely on short-term stimulation from ingredients like caffeine and taurine, lacking systemic intervention to regulate the root causes of energy metabolism and oxidative stress.

[0004] In recent years, probiotics have become a research hotspot for the prevention and treatment of metabolic diseases due to their intestinal microecological regulatory function. In the existing technology, although single or simply compounded probiotic preparations (such as Lactobacillus and Bifidobacterium) have been proven to relieve inflammation by inhibiting the secretion of proinflammatory factors (such as IL-1β, TNF-α), or indirectly affecting host metabolism by improving intestinal barrier function, their scope of action is mostly limited to the local intestine, and the regulatory effect on systemic inflammation is limited. At the same time, the existing probiotic combinations have insufficient effects on the key links of taurine metabolism, making it difficult to achieve long-term promotion of endogenous taurine metabolism.

[0005] Furthermore, the application of traditional probiotics in the anti-fatigue field focuses on alleviating post-exercise lactic acid accumulation or enhancing immunity, lacking multi-dimensional regulation of anti-inflammation, oxidative stress, taurine metabolism, and other aspects. Furthermore, the functional synergy of the strains is insufficient, resulting in unstable anti-fatigue effects. Therefore, the development of a compound black wolfberry probiotic composition that can synergistically combat inflammation, regulate taurine metabolism, and enhance the body's anti-fatigue ability has become an important direction for breaking through the bottleneck of existing technologies. Summary of the Invention

[0006] In order to overcome the above technical problems, the present invention provides an application of a composite black wolfberry probiotic composition in the preparation of anti-inflammatory, taurine metabolism-promoting and anti-fatigue products.

[0007] To achieve the above objectives, the technical solutions provided by the present invention are as follows:

[0008] A use of a composite black wolfberry probiotic composition in the preparation of an anti-inflammatory, and / or taurine anabolism promoting, and / or anti-fatigue product.

[0009] Preferably, the composite black wolfberry probiotic composition can reduce the level of inflammatory factor IL-6 in the body.

[0010] Preferably, the composition of the compound black wolfberry probiotic can reduce the level of TNF-α in the body.

[0011] Preferably, the composition of the compound black wolfberry probiotic can reduce the level of IL-1β in the body.

[0012] Preferably, the composite black wolfberry probiotic composition can promote taurine synthesis.

[0013] Preferably, the composite black wolfberry probiotic composition comprises, by weight: 30-60 parts of black wolfberry freeze-dried powder, 10-20 parts of oligofructose, and 1-5 parts of composite probiotic powder.

[0014] Preferably, the composite black wolfberry probiotic composition comprises, by weight: 30-50 parts of black wolfberry freeze-dried powder, 10-20 parts of oligofructose, and 1-5 parts of composite probiotic powder.

[0015] Preferably, the composite black wolfberry probiotic composition comprises, by weight: 35-35 parts of black wolfberry freeze-dried powder, 12-18 parts of oligofructose, and 2-4 parts of composite probiotic powder.

[0016] Preferably, the composite probiotic powder comprises Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis in a mass ratio of 4-6:2-4:1-3, and the total viable bacteria count of the composite probiotic powder is ≥1.5×10 11 CFU / g.

[0017] The preferred mass ratio of Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis is 5:3:2.

[0018] Preferably, the composite probiotic powder comprises Lactobacillus plantarum H6, Lactobacillus rhamnosus HN001, and Bifidobacterium animalis subspecies lactis HN019 in a mass ratio of 5:3:2, and the total viable bacteria count of the composite probiotic powder is ≥1.5×10 11 CFU / g.

[0019] Preferably, the method for preparing the black wolfberry freeze-dried powder comprises the following steps:

[0020] Soak wolfberry in water and beat it. After beating, let it stand, remove the floating matter on the liquid surface and add ascorbic acid. Use 0.5-1.5 mol / L citric acid solution to adjust the pH of black wolfberry juice to 4.0-5.0. Stir evenly and filter. Perform vacuum freeze-drying at -35-45℃, grind and pass through a 40-60 mesh sieve to obtain black wolfberry freeze-dried powder.

[0021] Preferably, the wolfberry is soaked in water and pulped. After the pulping is completed, the wolfberry is allowed to stand, and after removing the floating matter on the liquid surface, ascorbic acid is added. The pH of the black wolfberry juice is adjusted to 4.0-5.0 with 1 mol / L citric acid solution, and the mixture is stirred evenly and then filtered. The mixture is vacuum freeze-dried at -40°C, crushed, and sieved to obtain the black wolfberry freeze-dried powder.

[0022] Preferably, the mass of the added water is 10-25 times the mass of the wolfberry, the soaking time is 30-60 minutes, the standing time is 2-4 hours, the amount of ascorbic acid added is 0.05%-0.08% of the mass of the water, 1 mol / L citric acid solution is used to adjust the pH of the black wolfberry juice, the juice is stirred evenly and then filtered, vacuum freeze-dried at -40°C, crushed and sieved to a mesh size of 40-60 mesh.

[0023] On the other hand, the present invention provides an anti-fatigue product, the raw materials of which include the composite black wolfberry probiotic composition in the application and its food-acceptable auxiliary materials.

[0024] Compared with the prior art, the technical advantages of the present invention are:

[0025] On the one hand, the compound black wolfberry probiotic of the present invention metabolizes the active ingredients such as polyphenols and polysaccharides in black wolfberry to generate metabolites with stronger antioxidant capacity (aglycones, short-chain fatty acids, etc.). It can also significantly improve the anti-fatigue effect through multi-target synergistic antioxidant effects such as activating the Nrf2 pathway, scavenging free radicals, and enhancing taurine metabolism.

[0026] On the other hand, the short-chain fatty acids produced by the metabolism of the probiotics of the present invention can block the activity of histone deacetylase (HDAC), and black wolfberry polyphenols can inhibit IKKβ phosphorylation. The dual effects of the black wolfberry probiotic composition synergistically reduce NF-κB nuclear translocation and reduce the release of proinflammatory factors (IL-6, TNF-α and IL-1β), thereby breaking the vicious cycle of inflammatory factors and oxidative stress and alleviating fatigue symptoms.

[0027] In addition, the compound black wolfberry probiotic of the present invention can not only metabolize sulfur-containing compounds (such as cysteine) in black wolfberry to generate taurine precursors (such as hypotaurine), thereby promoting taurine anabolism; but also produce organic acids (such as lactic acid, acetic acid) by fermenting black wolfberry, and increase the content of organic acids (such as chlorogenic acid, caffeic acid and coumaric acid, etc.) by fermenting black wolfberry, which can not only better directly exert anti-inflammatory effects, but also lower intestinal pH, promote the activity of taurine synthase (such as cysteine sulfonic acid decarboxylase, CSAD), and promote taurine synthesis. The taurine generated can directly scavenge hydroxyl free radicals and peroxynitrite, inhibit lipid peroxidation, and reduce the release of pro-inflammatory factors (TNF-α, IL-6), so that the black wolfberry compound black wolfberry probiotic of the present invention can achieve anti-inflammatory and anti-fatigue effects through multiple pathways. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 is the concentration of IL-6, TNF-α and IL-1β in the serum of mice in different groups.

[0029] Figure 2 This figure shows the changes in various phenolic acids in the fermentation system of black wolfberry freeze-dried powder fermented with compound probiotics in vitro. For the same component, different letters represent significant statistical differences in the content of the component at different time points (p < 0.05). DETAILED DESCRIPTION

[0030] The present invention is described below by way of specific examples to make the technical solution of the present invention easier to understand and grasp, but the present invention is not limited thereto. The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials described are all commercially available unless otherwise specified.

[0031] Example 1 Anti-inflammatory and anti-fatigue test of the composition of the present invention compounded with black wolfberry probiotics

[0032] 1.1 Animal grouping

[0033] This experiment used 90 healthy male KM mice aged 6 weeks, purchased from Hangzhou Qizhen Experimental Animal Technology Co., Ltd. The mice were housed in the animal room of the Experimental Animal Center of East China University of Science and Technology with an ambient temperature of 25±2°C and a humidity of 50±5%. The mice were alternating between light and dark every 12 hours and had free access to food and water. This animal experiment was approved by the Bioethics Committee of East China University of Science and Technology, with approval number: ECUST-2025-003. After one week of adaptive feeding, the mice were randomly divided into a normal group, a model group, a salidroside group (drug positive control), a low-dose group of black wolfberry probiotic composition (referred to as the low-dose composition group), a high-dose group of black wolfberry probiotic composition (referred to as the high-dose composition group), a low-dose black wolfberry group, a high-dose black wolfberry group, a high-dose probiotic group, and a low-dose probiotic group according to their body weight, with 10 mice in each group.

[0034] Among them, the black wolfberry probiotic composition is calculated by weight as follows: 40 parts of black wolfberry freeze-dried powder, 15 parts of oligofructose, and 3 parts of compound probiotic powder.

[0035] The preparation method is as follows:

[0036] S1: Preparation of black wolfberry freeze-dried powder: First, remove bad fruits and impurities, select dry wolfberries with bright color and no physical damage, add 15 times the weight of deionized water, soak for 40 minutes and beat for treatment. After beating, let it stand for 3 hours, remove the floating matter on the liquid surface and add 0.06% ascorbic acid, adjust the pH of the black wolfberry juice to 4.5 with 1 mol / L citric acid solution, stir evenly and filter, vacuum freeze-dry at -40℃, crush, and pass through a 60-mesh sieve to obtain black wolfberry freeze-dried powder.

[0037] S2: The material ratio of black wolfberry freeze-dried powder, oligofructose, and probiotic compound powder (Lactobacillus plantarum H6, Lactobacillus rhamnosus HN001, and Bifidobacterium animalis subsp. lactis HN019) is 5:3:2, and the total viable bacteria count of the compound powder is ≥1.5×10 11 CFU / g) are fully mixed and subjected to metal detection to obtain the black wolfberry probiotic composition of the present invention.

[0038] Among them, Lactobacillus plantarum was purchased from but not limited to Shan En Kang Biotechnology (Suzhou) Co., Ltd.; Lactobacillus rhamnosus and Bifidobacterium animalis were obtained from but not limited to Danisco Biotechnology (Shanghai) Co., Ltd. The compound probiotics can also be purchased from Shan En Kang Biotechnology (Suzhou) Co., Ltd., product batch number W2406080.

[0039] 1.2 Insomnia and inflammation mouse model establishment and drug administration

[0040] Three days before the formal experiment, a sodium pentobarbital suprathreshold dose reversal experiment was performed. Except for the normal group, mice in all groups received intraperitoneal injections of 300 mg / kg parachlorophenylalanine (PCPA) starting on day 1 for two consecutive days. The sodium pentobarbital suprathreshold dose reversal experiment was repeated on day 3. Serum levels of inflammatory factors (IL-6, TNF-α, and IL-1β) were measured and compared with pre-modeling serum levels of IL-6, TNF-α, and IL-1β in the same mice to assess modeling efficacy. PCPA was administered intraperitoneally again on day 10 to consolidate the modeling effect. The drug-treated groups received daily gavage from day 1 to day 14. Serum levels of inflammatory factors (TNF-α, IL-6, and IL-1β) were measured 10 minutes after gavage on day 14. A forced swim test was performed on day 15, with the duration of exhaustive swimming used as an indicator of drug efficacy against fatigue. The criterion for exhaustion was that the mouse sank to the bottom of the water for 10 seconds without floating. Table 1 shows the dosing methods for mice in each group.

[0041] Table 1

[0042]

[0043] 1.3 Data Analysis

[0044] All numerical values in the experiment are expressed as (mean ± standard deviation). The results were compared with the differences between different groups by one-way analysis of variance (ANOVA) using the statistical software GraphPadPrism 10, and the significance level was set at p < 0.05.

[0045] 1.4 Experimental Results

[0046] (1) The effects of the composition on IL-6, TNF-α, IL-1β inflammatory factors in mouse serum and forced swimming are shown in Table 2 and Figure 1 .

[0047] Table 2

[0048]

[0049]

[0050] As shown in Table 2 and Figure 1 As shown in the figure, the IL-6 concentration in the model group was significantly higher than that in the normal group (*p<0.05), indicating that the model group had an inflammatory response. The IL-6 concentrations in the salidroside group and the high-dose black wolfberry probiotic composition group were significantly lower than those in the model group (*p<0.05, **p<0.01), while the IL-6 concentration in the low-dose composition group was not significantly different from that in the model group.

[0051] As shown in Table 2 and Figure 1As shown in the figure, the TNF-α concentration in the model group was significantly higher than that in the normal group (*p<0.05), which also indicated that the model group had an inflammatory response. The TNF-α concentration in the high-dose group was significantly lower than that in the model group (**p<0.01), and the TNF-α concentration in the low-dose combination group was also lower than that in the model group (*p<0.05).

[0052] As shown in Table 2 and Figure 1 As shown, the IL-1β concentration in the model group was significantly higher than that in the normal group (*p<0.05), indicating the presence of an inflammatory response. The IL-1β concentration in the high-dose combination group was similar to that in the normal group and significantly lower than that in the model group (**p<0.01), indicating that the high-dose combination group helped to alleviate inflammation. The IL-1β concentration in the low-dose combination group also showed a significant difference compared with the model group (*p<0.05), but was higher than that in the high-dose combination group.

[0053] As shown in Table 2, the exhaustive swimming time of the model group was significantly lower than that of the normal group (*p<0.05), indicating that the model group had more obvious fatigue symptoms. The high-dose group of the composition significantly increased the exhaustive swimming time compared with the model group (*p<0.05), indicating that it had a significant anti-fatigue effect.

[0054] The increase in IL-6, TNF-α, and IL-1β concentrations in the model group may be due to the activation of the inflammatory response. As an antidepressant, salidroside may exert its therapeutic effect by inhibiting the inflammatory response in addition to its effect on the neurotransmitter system. The effect of the high-dose group of the black wolfberry probiotic composition in reducing the concentrations of IL-6, TNF-α, and IL-1β in the serum indicates that the composition has an anti-inflammatory effect, which may be one of the mechanisms of its antidepressant and fatigue-relieving effects. However, the effect of the low-dose group was also significant (*p<0.05), indicating that dosage may be one of the key factors affecting its efficacy.

[0055] 1.5 The present invention further tested the taurine content in the serum of each group of mice. The results are shown in Table 3.

[0056] Table 3

[0057]

[0058]

[0059] Compared with the normal group, the model group *p<0.05, compared with the model group, *p<0.05.

[0060] In addition to the above experiments, the present invention studied the changes in the content of organic acids, total flavonoids, total phenols, and anthocyanins in the fermentation system of black wolfberry freeze-dried powder fermented by a compound probiotic in vitro. The changes in phenolic acid are shown in Figure 2. Figure 2The contents of chlorogenic acid, caffeic acid, and p-coumaric acid continued to increase with the extension of fermentation time; the increase of chlorogenic acid may be due to the combination of free hydroxycinnamic acid with quinic acid in the food matrix; the increase of caffeic acid may be due to the cinnamoyl esterase produced by lactic acid bacteria decomposing the conjugated cinnamic acid in the black wolfberry matrix to obtain free caffeic acid.

[0061] The total flavonoid content in this experiment was 1.90 mg / mL before fermentation. The flavonoid content in the fermentation broth increased significantly after fermentation (p<0.05). The total flavonoid content in the fermentation broth after 4 h was the highest, reaching 2.59 mg / mL (an increase of 36.32%).

[0062] Before fermentation, the total phenol content in black wolfberry was 2.77 mg / mL. After 4 hours of fermentation, the total phenol content in the fermentation broth increased significantly. There was no significant difference in total phenol content between 4 and 24 hours of fermentation with the composite probiotics of the present invention, with the highest total phenol content reaching 3.21 mg / mL after 24 hours of fermentation. This represents a 15.89% increase after fermentation.

[0063] The total anthocyanin content in black wolfberry juice was 3.762 mg / mL before fermentation. After fermentation, the total anthocyanin content increased significantly (p < 0.05). Throughout the fermentation process, the content initially increased, then decreased, and then stabilized. The highest total anthocyanin content reached 6.67 mg / mL after 4 hours of fermentation.

[0064] The present invention has found that the composite probiotics and black wolfberry of the present invention can metabolize the active ingredients such as polyphenols, polysaccharides, total flavonoids in black wolfberry, or significantly increase the content of their active ingredients, or generate metabolites with stronger antioxidant capacity (aglycones, short-chain fatty acids, etc.). It can significantly improve the anti-fatigue effect through multi-target synergistic antioxidant effects such as activating the Nrf2 signaling pathway, scavenging free radicals, and enhancing taurine metabolism.

[0065] By fermenting black wolfberry to produce organic acids (such as lactic acid and acetic acid), and by fermenting black wolfberry to increase its organic acid content (such as chlorogenic acid, caffeic acid and coumaric acid, etc.), not only can the anti-inflammatory effect be better exerted directly, but also the intestinal pH value can be reduced, the activity of taurine synthase (such as cysteine sulfenic acid decarboxylase, CSAD) can be promoted, and taurine synthesis can be promoted. The taurine produced can directly scavenge hydroxyl free radicals and peroxynitrite, inhibit lipid peroxidation, and reduce the release of pro-inflammatory factors (TNF-α, IL-6), so that the black wolfberry compound black wolfberry probiotic of the present invention can achieve anti-inflammatory and anti-fatigue effects through multiple pathways.

[0066] The above detailed description is a specific description of one feasible embodiment of the present invention. This embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not depart from the present invention should be included in the scope of the technical solution of the present invention.

Claims

1. Use of a composite black wolfberry probiotic composition in the preparation of anti-inflammatory, taurine metabolism-promoting and anti-fatigue products.

2. The use according to claim 1, characterized in that The composite black wolfberry probiotic composition can reduce the level of inflammatory factor IL-6 in the body.

3. The use according to claim 1, characterized in that The composite black wolfberry probiotic composition can reduce the level of TNF-α in the body.

4. The use according to claim 1, characterized in that The composite black wolfberry probiotic composition can reduce the level of IL-1β in the body.

5. The use according to claim 1, characterized in that The composite black wolfberry probiotic composition can promote taurine metabolism.

6. The use according to claim 1, characterized in that The composite black wolfberry probiotic composition comprises, by weight, 30-60 parts of black wolfberry freeze-dried powder, 10-20 parts of oligofructose, and 1-5 parts of composite probiotic powder.

7. The use according to claim 6, characterized in that The composite probiotic powder comprises Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium animalis in a mass ratio of 5:3:2, and the total viable bacterial count of the composite probiotic powder is ≥1.5×10 11 CFU / g.

8. The use according to claim 6, characterized in that The preparation method of the black wolfberry freeze-dried powder comprises the following steps: soaking wolfberries in water, beating the wolfberries, allowing the beating to stand, removing the floating matter on the liquid surface, adding ascorbic acid, adjusting the pH of the black wolfberry juice to 4.0-5.0 with a 0.5-1.5 mol / L citric acid solution, stirring evenly, filtering, vacuum freeze-drying at -35-45°C, crushing, and passing through a 40-60 mesh sieve to obtain the black wolfberry freeze-dried powder.

9. The use according to claim 8, characterized in that The mass of the added water is 10-25 times the mass of the wolfberry, the soaking time is 30-60 minutes, the standing time is 2-4 hours, the amount of ascorbic acid added is 0.05%-0.08% of the mass of the water, 1 mol / L citric acid solution is used to adjust the pH of the black wolfberry juice, the juice is stirred evenly and then filtered, vacuum freeze-dried at -40°C, crushed and sieved to a mesh size of 40-60 mesh.

10. An anti-fatigue product, characterized in that: The raw materials of the anti-fatigue product include the composite black wolfberry probiotic composition according to any one of claims 1 to 9 and its food-acceptable auxiliary materials.

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