Composition with anti-inflammatory function as well as preparation method and application thereof

Abstract

The invention provides a composition with an anti-inflammatory function as well as a preparation method and application thereof, and relates to the technical field of dietary nutrition supplements for improving inflammation. The composition with the anti-inflammatory function is prepared from hucho taimen oil and ergothioneine. The composition provided by the invention not only can comprehensively inhibit cardiovascular inflammatory reaction through multiple mechanisms, but also can improve the stability and delivery efficiency of the product, and provides important theoretical and practical basis for developing novel and efficient anti-inflammatory dietary supplement products.

Description

Technical Field

[0001] The present invention relates to the technical field of dietary nutritional supplements for improving inflammation, and more particularly, to a composition having anti-inflammatory functions, a preparation method thereof, and an application thereof. Background Art

[0002] Vascular diseases, including atherosclerosis, myocardial infarction, etc., are one of the main causes of death and disability worldwide. The occurrence and development of these diseases are closely related to chronic inflammatory responses. The overexpression of inflammatory factors such as IL-6, TNF-α, and NF-κB plays a key role in cardiovascular injury and disease progression. Therefore, anti-inflammation and improvement of chronic inflammation have become one of the important strategies for cardiovascular disease intervention. In recent years, dietary nutritional supplements have gradually attracted attention as an auxiliary means.

[0003] In the field of dietary nutritional supplements, most of the existing anti-inflammatory products focus on the supplementation of single components, such as fish oil, curcumin, or vitamin D. Although these components have certain anti-inflammatory effects, they often can only intervene in a single inflammatory factor or pathway, and it is difficult to comprehensively inhibit complex inflammatory responses. In addition, many dietary supplements have deficiencies in bioavailability and delivery efficiency, and cannot effectively deliver active ingredients to the inflammatory site, thus limiting their anti-inflammatory effects. At the same time, some supplements may not be able to fully exert their potential effects due to unreasonable formulations or lack of synergistic effects.

[0004] The improvement and treatment of cardiovascular inflammation require supplements to be accurately delivered to the inflammatory site. However, in the existing technology, the lack of targeting of supplements is an urgent problem to be solved. Although nano-delivery systems have made some progress in improving stability and delivery efficiency, these systems are still difficult to achieve precise targeting of cardiovascular inflammatory sites. In addition, cardiovascular inflammation involves multiple inflammatory factors and cell signaling pathways, and single-component supplements often cannot comprehensively inhibit inflammatory responses. In the existing technology, although there are studies attempting to deliver multiple inflammation-inhibiting and improving components through nanotechnology, the synergistic effects between these components are not ideal, and it is difficult to achieve the expected therapeutic effects.

[0005] In summary, the existing anti-inflammatory dietary nutritional supplements have many defects in improving cardiovascular inflammation, including limited effects of single components, low bioavailability and delivery efficiency, lack of targeting, and unsatisfactory synergistic anti-inflammatory effects. These problems not only limit the application effects of anti-inflammatory supplements in the prevention, treatment, and improvement of cardiovascular diseases, but also pose an urgent need for the development of more efficient and precise anti-inflammatory dietary supplements.

[0006] In view of this, the present invention is specifically proposed. Summary of the Invention

[0007] The object of the present invention is to provide a composition with anti-inflammatory function, its preparation method and application. The composition with anti-inflammatory function can not only comprehensively reduce cardiovascular inflammatory responses through multiple mechanisms, but also improve the stability and delivery efficiency of the composition, providing an important theoretical and practical basis for the development of new and efficient cardiovascular anti-inflammatory dietary nutritional supplements.

[0008] In order to achieve the above object of the present invention, the following technical solutions are specifically adopted: In the first aspect, the present invention provides a composition with anti-inflammatory function, including: Tisbe fasciata oil and ergothioneine.

[0009] In an alternative embodiment, in the composition with anti-inflammatory function, the weight ratio of Tisbe fasciata oil to ergothioneine is 100:(1 - 10).

[0010] In an alternative embodiment, it further includes an emulsifier.

[0011] In an alternative embodiment, the emulsifier includes beeswax or lecithin.

[0012] In an alternative embodiment, the addition amount of the emulsifier accounts for 0.5% - 1.5% of the total amount of the composition with anti-inflammatory function.

[0013] In an alternative embodiment, the composition with anti-inflammatory function is an oil-in-water emulsion composed of ergothioneine coated with nanoparticles and Tisbe fasciata oil.

[0014] In an alternative embodiment, the nanoparticles include nanoliposomes and / or polymer nanoparticles.

[0015] In the second aspect, the present invention provides a preparation method of the composition with anti-inflammatory function according to any one of the foregoing embodiments, including: Mix Tisbe fasciata oil and ergothioneine to obtain the composition with anti-inflammatory function.

[0016] In an alternative embodiment, the step of mixing Tisbe fasciata oil and ergothioneine to obtain the composition with anti-inflammatory function includes: Add Tisbe fasciata oil and ergothioneine into a container and stir to mix to obtain a mixture; Add an emulsifier to the mixture and continue to stir to mix to obtain the composition with anti-inflammatory function.

[0017] In an alternative embodiment, the particle size of ergothioneine is not less than 120 mesh; and / or, the emulsifier is beeswax or lecithin; and / or, the addition amount of the emulsifier is 0.5% - 1.5% of the total amount of the composition with anti-inflammatory function.

[0018] In a third aspect, the present invention provides an application of the composition according to any one of the foregoing embodiments in the preparation of a dietary nutritional supplement for improving chronic cardiovascular inflammation.

[0019] The present invention provides a composition with anti-inflammatory function, its preparation method and application. Among them, the composition with anti-inflammatory function exhibits significant synergistic effects through the combination of Calanus finmarchicus oil and ergothioneine. Calanus finmarchicus oil is rich in various bioactive components, such as long-chain fatty acids (including EPA, DHA), astaxanthin, and exists in the form of wax esters, etc. These components have effects such as antioxidant, anti-inflammatory, and cardiovascular protection. As a natural antioxidant, ergothioneine can effectively scavenge free radicals, reduce oxidative stress, and inhibit the overexpression of inflammatory factors. The synergistic effect between the two lies in that astaxanthin and long-chain fatty acids in Calanus finmarchicus oil can enhance the antioxidant ability of ergothioneine, while ergothioneine can further enhance the anti-inflammatory effect of Calanus finmarchicus oil. This synergistic effect not only improves the anti-inflammatory and antioxidant effects of single components, but also inhibits the production of inflammatory factors (such as IL-6, TNF-α) through a multi-target mechanism, regulates the NF-κB signaling pathway, thereby more comprehensively blocking the cardiovascular inflammatory response. In addition, the oil matrix of Calanus finmarchicus oil provides a good carrier for ergothioneine, improves its stability, and further enhances the delivery efficiency of the composition. This synergistic effect provides a more efficient and stable solution for the prevention and improvement of cardiovascular diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0021] Figure 1 It is a columnar result diagram of cell viability in Test Experimental Example 2 of this application; Figure 2 It is a result diagram of the cell apoptosis experiment in Test Experimental Example 3 of this application; Figure 3 It is a result diagram of the inflammatory factor IL-6 in Test Experimental Example 4 of this application; Figure 4 It is a result diagram of the inflammatory factor TNF-α in Test Experimental Example 4 of this application; Figure 5 It is a result diagram of the inflammatory factor NF-κB in Test Experimental Example 4 of this application. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0022] The embodiments of the present invention will be described in detail below in conjunction with examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. For those not specified in the examples, the operations are carried out under conventional conditions or conditions recommended by the manufacturer. All reagents or instruments not indicated by the manufacturer can be obtained as conventional products through commercial channels.

[0023] In an embodiment of the present application, a composition with anti-inflammatory function is provided, including: Calanus finmarchicus oil and ergothioneine.

[0024] Calanus finmarchicus oil (also known as red jewel oil) is a unique marine oil extracted from the zooplankton Calanus finmarchicus. It is rich in various bioactive components, such as wax esters, long-chain fatty acids (including EPA and DHA), and astaxanthin, etc. These components endow Calanus finmarchicus oil with multiple effects such as antioxidant, anti-inflammatory, and cardiovascular protection.

[0025] Ergothioneine (EGT) is a naturally occurring sulfur-containing amino acid derivative, first isolated from ergot fungi. It is a water-soluble antioxidant and widely exists in mushrooms, animal livers, kidneys, and certain legumes. Since the human body cannot synthesize ergothioneine by itself, it must be obtained through diet or external supplementation to meet physiological needs. Ergothioneine has multiple biological activities, including scavenging free radicals, protecting mitochondria and DNA inside cells, and maintaining the normal growth and immune function of cells.

[0026] The combination of ergothioneine and Calanus finmarchicus oil not only gives full play to their respective advantages but also enhances the overall anti-inflammatory and antioxidant effects through synergistic effects, providing a new solution for the prevention and treatment of cardiovascular diseases.

[0027] In the treatment of cardiovascular inflammation, most existing anti-inflammatory dietary supplements or anti-inflammatory products only target a single inflammatory factor or pathway and are difficult to comprehensively inhibit complex inflammatory reactions. However, the composition of Calanus finmarchicus oil and ergothioneine can act on multiple inflammation-related targets simultaneously, including inhibiting the overexpression of inflammatory factors (such as IL-6, TNF-α) and regulating the activity of the NF-κB signaling pathway, thereby more effectively blocking the progression of the inflammatory reaction. In addition, this composition can also significantly improve cell viability, reduce apoptosis, protect cardiovascular endothelial cells from inflammatory damage, and thus maintain the normal function of the cardiovascular system.

[0028] From the perspective of drug delivery, the lipid matrix of Calanus finmarchicus oil provides a good carrier for ergothioneine, which helps to improve the stability and bioavailability of ergothioneine. This composition not only solves the problems of easy degradation and poor stability of ergothioneine in water-soluble media, but also achieves a more efficient delivery effect by optimizing the ratio of the two, enabling the active ingredient to act more precisely on the cardiovascular inflammation site and improving the therapeutic effect.

[0029] In summary, the composition of Calanus finmarchicus oil and ergothioneine shows significant advantages in the field of cardiovascular anti-inflammation. It can not only comprehensively inhibit the cardiovascular inflammatory response through multiple mechanisms, but also improve the stability and delivery efficiency of the composition, providing an important theoretical and practical basis for the development of new and efficient cardiovascular anti-inflammatory products (such as dietary supplement products).

[0030] In some embodiments, in the composition having anti-inflammatory function, the weight ratio of Calanus finmarchicus oil to ergothioneine is 100:(1~10). For example, it can be 100:1, 100:2, 100:5, 100:7, 100:10, etc.

[0031] In some embodiments, it further includes an emulsifier.

[0032] In some embodiments, the emulsifier includes beeswax or lecithin.

[0033] In some embodiments, the addition amount of the emulsifier accounts for 0.5%~1% of the total amount of the composition having anti-inflammatory function. For example, it can be 0.5%, 0.7%, 0.9%, 1.2%, 1.5%, etc.

[0034] It should be noted that an emulsifier is a surfactant with hydrophilic and lipophilic groups. It can reduce the interfacial tension between the oil phase and the water phase, enabling two immiscible liquids to form a stable emulsion. In the composition of Calanus finmarchicus oil (oil phase) and ergothioneine (usually water-soluble), the emulsifier helps these two components to mix and disperse better through its hydrophilic and lipophilic properties.

[0035] In the composition of Calanus sinicus oil and ergothioneine, the functions of the emulsifier are mainly reflected in the following aspects: First, the emulsifier can reduce the interfacial tension between oil and water, enabling the oil-soluble Calanus sinicus oil and the water-soluble ergothioneine to form a stable emulsion, thereby achieving uniform dispersion and solving the problem of difficult mixing in their natural state. Second, the emulsifier forms a protective film at the oil-water interface to prevent droplet aggregation and separation, significantly improving the stability of the composition, especially playing an important role in preventing precipitation and stratification, which is crucial for the stability of the composition during storage and use. In addition, through emulsification, the emulsifier disperses Calanus sinicus oil and ergothioneine into smaller particles, increasing their contact area with biological membranes, thereby enhancing the bioavailability of both and promoting absorption and efficacy in the body. Finally, the emulsifier can also make the composition form a uniform emulsion, improving the appearance and texture of the product, which is particularly important for the application of the composition in foods, health products, or cosmetics, as a uniform appearance and good texture can significantly enhance the attractiveness and user experience of the product.

[0036] In some embodiments, the composition with anti-inflammatory function is an oil-in-water emulsion composed of ergothioneine coated with nanoparticles and Calanus sinicus oil.

[0037] As mentioned above, ergothioneine is a water-soluble substance. Although ergothioneine has good water solubility, its gastrointestinal absorption rate is still slow, especially the solubility in the gastrointestinal tract and the permeability of the composition through cell membranes are not ideal.

[0038] In this embodiment, in order to improve the absorption rate of ergothioneine, by encapsulating the water-soluble substance (ergothioneine) in nanoparticles, the surface area of the substance can be significantly increased, its solubility can be improved, and its absorption in the gastrointestinal tract can be accelerated. In this embodiment, the nanoparticles can be polymer nanoparticles, nanoliposomes, etc. The mechanism is that nanoparticles can accelerate the dissolution process of the substance by increasing its surface area, thereby improving the bioavailability of the target substance.

[0039] It should be noted that encapsulating ergothioneine in nanoparticles does not affect its absorption, but may instead improve its absorption efficiency. The actual effect depends on factors such as the type, size, and surface modification of the nanoparticles. In this case, nanoparticles can promote the absorption of ergothioneine by increasing the surface area, improving solubility, and enhancing cell membrane penetration, especially in the gastrointestinal tract.

[0040] Although ergothioneine itself is water-soluble, nanoparticles can further improve its solubility and bioavailability. By encapsulating ergothioneine within nanoparticles, the surface area of the substance can be significantly increased, thereby accelerating its dissolution in the gastrointestinal tract, especially for substances with poor solubility. The size of nanoparticles is typically in the nanoscale (usually 50 - 200 nanometers), which greatly increases the surface area of the substance, promotes the dissolution of the substance in the aqueous phase, and thus accelerates absorption. By dispersing ergothioneine within nanoparticles, the dissolution rate of the substance can be increased, enabling it to dissolve and be absorbed more rapidly in the gastrointestinal tract.

[0041] Through nanoparticle encapsulation, the release of the substance can be precisely controlled. This means that the substance will not be metabolized or excreted too quickly in the body, thus maintaining an effective concentration for a longer time. Nanoparticles provide a mechanism for the slow release of active substances. By controlling the release rate of the active substance, the rapid release and absorption of the active substance are avoided, enabling the active substance to maintain an effective concentration for a longer time and achieving a better therapeutic effect. Since nanoparticles can protect the active substance and reduce its degradation in the body, the half-life of the active substance is prolonged, thereby increasing its bioavailability.

[0042] The morphology and size of nanoparticles enable them to more easily pass through cell membranes, increasing the chance of active substances entering cells. Liposomes or polymer nanoparticles have physical properties similar to cell membranes, which allows them to facilitate the penetration of active substances through cell membranes, thereby accelerating the absorption of active substances.

[0043] It should be noted that chronic cardiovascular inflammation is a long-term pathological process that requires continuous anti-inflammatory and antioxidant treatment. As a potent antioxidant and anti-inflammatory agent, ergothioneine can be slowly released in the body through sustained-release technology, extending the effective action time of the active substance. This continuous release of the active substance can better maintain the concentration of ergothioneine in the body, avoiding unstable therapeutic effects caused by fluctuations in the concentration of the active substance.

[0044] The sustained-release system can reduce the frequency of drug administration, thereby improving patient compliance. In addition, by controlling the release rate, the instantaneous concentration of ergothioneine in the body can be reduced, minimizing potential side effects such as gastrointestinal discomfort or liver and kidney burden. This is particularly important for patients with chronic cardiovascular inflammation.

[0045] Therefore, although nanoparticles themselves can already improve the stability and release characteristics of active substances, under the premise of aiming for sustained release, further treatment using water-in-oil (W / O) emulsions may bring additional advantages: First, the water-in-oil emulsion system can further slow down the release of the composition. In the water-in-oil system, the oil phase acts as the continuous phase and is encapsulated in the aqueous core or the oil phase, which can provide a longer-lasting sustained release effect, especially for active substances that require a long time to exert their therapeutic effects. Second, the oil phase in the water-in-oil emulsion can stabilize substances and protect them from environmental factors (such as oxidation, moisture, etc.). Especially when the active substance needs to be stored for a long time or stably exist in the gastrointestinal tract, the oil phase helps prevent the degradation of the active ingredient. Third, if the composition requires specific targeted delivery (for example, targeting the cardiovascular system), using a water-in-oil emulsion can better carry the active substance to the target area and control the release through the oil phase, thereby enhancing the targeting and effectiveness of the active substance.

[0046] In some embodiments, the nanoparticles include nanoliposomes and / or polymer nanoparticles.

[0047] Nanoliposomes are vesicle structures composed of a phospholipid bilayer and usually contain one or more aqueous cores. Its main components include phospholipids, cholesterol, and polyethylene glycol-modified lipids, etc. The diameter of nanoliposomes is generally about 100 nanometers, and it has good biocompatibility and biodegradability, and can effectively encapsulate and protect active substance molecules, preventing their degradation and premature release in the body.

[0048] Polymer nanoparticles are nanoparticles made of synthetic or natural polymers. Its preparation methods include emulsion polymerization, microemulsion polymerization, and self-assembly methods, etc. The size of polymer nanoparticles is usually between 10 - 1000 nanometers, and it has good stability and adjustable active substance release ability.

[0049] In the embodiments of the present application, a preparation method of a composition having an anti-inflammatory function as described in any one of the foregoing embodiments is provided, including: Step S100, mixing Tisbe furcata oil and ergothioneine to obtain the composition having an anti-inflammatory function.

[0050] In some embodiments, the mixing of Tisbe furcata oil and ergothioneine to obtain the composition includes: Step S1, adding Tisbe furcata oil and ergothioneine into a container and stirring and mixing to obtain a mixture; Step S2, adding an emulsifier to the mixture and continuing to stir and mix to obtain the composition having an anti-inflammatory function.

[0051] In some embodiments, the particles of ergothioneine are not less than 120 mesh.

[0052] In some embodiments, the emulsifier is beeswax or lecithin.

[0053] In some embodiments, the addition amount of the emulsifier is 0.5% - 1% of the total amount of the composition having anti-inflammatory function. For example, it can be 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, etc.

[0054] In the embodiments of the present application, there is provided an application of a composition having anti-inflammatory function as described in any one of the foregoing embodiments in the preparation of an anti-inflammatory dietary supplement product.

[0055] As mentioned above, the anti-inflammatory dietary supplement product can be a dietary supplement for anti-chronic cardiovascular inflammation.

[0056] The present invention will be further described below through specific examples. However, it should be understood that these examples are only for more detailed description and should not be construed as limiting the present invention in any form.

[0057] Example 1 In this example, a composition having anti-inflammatory function was prepared.

[0058] Experimental method: Weigh 10 g and 1000 g respectively according to the ratio of ergothioneine (120-mesh fine powder): Thalassiosira weissflogii oil = 1:100, add them to a stirring device, and stir at a rotation speed of <100 r / min for 10 min.

[0059] After stirring, add 0.5% phospholipid (i.e., 5.05 g), increase the stirring speed to 500 r / min, and stir for 30 min until evenly mixed to obtain the composition.

[0060] Example 2 In this example, a composition having anti-inflammatory function was prepared.

[0061] Experimental method: Weigh 30 g and 1000 g respectively according to the ratio of ergothioneine (120-mesh fine powder): Thalassiosira weissflogii oil = 3:100, add them to a stirring device, and stir at a rotation speed of <100 r / min for 10 min.

[0062] After stirring, add 0.5% phospholipid (i.e., 5.15 g), increase the stirring speed to 500 r / min, and stir for 30 min until evenly mixed to obtain the composition.

[0063] Example 3 In this example, a composition having anti-inflammatory function was prepared.

[0064] Experimental methods: Weigh 50 g and 1000 g of ergothioneine (120 mesh micropowder) and Pegasus Daphnia oil in a ratio of 5:100, add them into a stirring device, and stir at a speed of <100 r / min for 10 min.

[0065] After stirring, 1% phospholipid (i.e., 10.5 g) was added, the stirring speed was increased to 500 r / min, and the mixture was stirred for 30 minutes until it was uniformly mixed to obtain a composition.

[0066] Example 4 In this example, a composition having anti-inflammatory function was prepared.

[0067] Experimental methods: Weigh 50 g and 1000 g of ergothioneine (120 mesh micropowder) and Pegasus Daphnia oil in a ratio of 5:100, add them into a stirring device, and stir at a speed of <100 r / min for 10 min.

[0068] After stirring, 0.5% beeswax (ie, 5.25 g) was added, the stirring speed was increased to 500 r / min, and the mixture was stirred for 30 minutes until the mixture was uniformly mixed to obtain a composition.

[0069] Comparative Example 1 In this comparative example, a composition was prepared.

[0070] Experimental methods: Weigh 50 g and 1000 g of ergothioneine (60 mesh): Pegasus Daphnia oil in a ratio of 5:100, add them into a stirring device, stir at a speed of <100 r / min for 10 min, and let stand.

[0071] Comparative Example 2: In this comparative example, a composition was prepared.

[0072] Experimental methods: Weigh 50 g and 1000 g of ergothioneine (120 mesh): Daphnia pulex oil in a ratio of 5:100, add them into a stirring device, and stir at a speed of <100 r / min for 10 min.

[0073] After stirring, 0.1% beeswax (ie, 1.05 g) was added, the stirring speed was increased to 500 r / min, and the mixture was stirred for 30 minutes until the mixture was uniformly mixed, thereby obtaining a comparative composition.

[0074] Test Example 1 In this test experimental example, the preparation processes of the compositions of different examples and comparative examples were investigated.

[0075] Experimental method: The samples prepared in Examples 1-4, Comparative Example 1 and Comparative Example 2 were placed for different times, and the states of the samples were observed.

[0076] Experimental results: Table 1. Observation results of the standing states of examples and comparative examples

[0077] Referring to the results in Table 1, by increasing ergothioneine at a specific mesh number and adding a specific proportion of emulsifier, the stability of the composition can be significantly improved.

[0078] Test experimental example 2 In this test experimental example, the viability of vascular endothelial cells of the composition was investigated.

[0079] Experimental method: Grouping: blank group, model group, ergothioneine group, Paracyclopina nana oil group, composition group (the composition in Example 2, i.e., the combination of ergothioneine + Paracyclopina nana oil); The cell viability was detected by the MTT method. Cells were seeded in 96-well plates at a density of 3000 cells per well. After the cells in each group were treated with the test substances for 24 h, the old culture medium was discarded. 100 μL of MTT at 0.5 g·L -1 was added to each well. After 4 h, the supernatant was discarded. 150 μL of DMSO was added to each well, and it was shaken for 10 min to dissolve. The OD value was measured at a wavelength of 490 nm using a multifunctional microplate reader, and the cell viability was calculated.

[0080] Experimental results: Table 2. Cell viability

[0081] Note: When performing statistical analysis using the t-test two-tailed test method, when comparing the model group with the blank group, the significance is indicated by #, p < 0.05 is indicated by #, p < 0.01 is indicated by ##. When comparing the sample group with the model group, the significance is indicated by *, p < 0.05 is indicated by *, p < 0.01 is indicated by **.

[0082] Reference Figure 1 (The red precious oil in the figure is Paracyclopina nana oil) and Table 2, it can be seen that compared with the single ergothioneine group and the single Paracyclopina nana oil group in terms of the improvement of cell viability, the composition can significantly (p < 0.01) improve the viability of vascular endothelial cells, indicating that the composition has an outstanding effect on the protection of the vascular endothelium.

[0083] Test Experiment Example 3 In this test experiment example, an apoptosis experiment of vascular endothelial cells of the composition was conducted.

[0084] Experimental method: Grouping: blank group, model group, ergothioneine group, Calanus sinicus oil group, composition group (the composition in Example 2, i.e., the combination of ergothioneine + Calanus sinicus oil); For the apoptosis experiment of HUVECs cells, palmitic acid was selected for modeling.

[0085] Cells were inoculated into 6-well plates at a density of 5×10 4 cells per well. After the cells adhered for 24 hours, the old culture medium was discarded. After treating the cells with the test substances according to the grouping, they were continuously cultured in an incubator for 24 hours, and then the old culture solution was discarded. The cells were washed twice with PBS, stained with Hoechst 33342 staining solution, and photographed under an inverted fluorescence microscope.

[0086] Experimental results: Reference Figure 2 , compared with the blank group, the nuclei in the model group had more fragmented shapes, the nuclei were shrunken, and the nuclear morphology was irregular, indicating that apoptosis occurred in the cells.

[0087] Compared with the model group, after the combination of Calanus sinicus oil, ergothioneine, and the combination of Calanus sinicus oil & ergothioneine, the phenomena of nuclear fragmentation, nuclear shrinkage, and irregular morphology were all reduced. Among them, the effect of the combination group of Calanus sinicus oil & ergothioneine was more significant, indicating that the combination of the two can produce a better anti-apoptotic effect.

[0088] Test Experiment Example 4 In this test experiment example, a test experiment on inflammatory factors of vascular endothelial cells of the composition was conducted.

[0089] Experimental method: Investigate inflammatory factor indicators: IL-6, TNF-α, and NF-κB.

[0090] Grouping: blank group, model group, ergothioneine group, Calanus sinicus oil group, composition group (the composition in Example 2, i.e., the combination of ergothioneine + Calanus sinicus oil); Collect the cell culture medium, perform experimental operations according to the instructions of the detection kit, and detect IL-6, TNF-α, and NF-κB in the cell culture medium by ELISA method.

[0091] Experimental results: Table 3. Results of the content of inflammatory factor IL-6

[0092] Note: When using the t-test two-tailed test method for statistical analysis, when comparing the model group with the blank group, the significance is indicated by #, p < 0.05 is indicated by #, p < 0.01 is indicated by ##, when comparing the sample group with the model group, the significance is indicated by *, p < 0.05 is indicated by *, p < 0.01 is indicated by ** (the same below).

[0093] Table 4. Results of the content of inflammatory factor TNF-α

[0094] Table 5. Results of the content of inflammatory factor NF-κB

[0095] Reference Figure 3 、 Figure 4 and Figure 5 (the red precious oil in the figure is the calanus sinicus oil), and the results in Table 3, Table 4 and Table 5, it can be seen that the composition has extremely significant improvement effects on the three endothelial cell inflammation indexes NF-κB, TNF-α, and IL-6 induced by palmitic acid.

[0096] In Test Example 4, the contents of inflammatory factors IL-6, TNF-α and NF-κB were investigated to evaluate the anti-inflammatory effect of the composition group (ergothioneine + calanus sinicus oil). Reference Figure 3 、 Figure 4 and Figure 5 , the experimental results show that the composition group is significantly superior to the group using only ergothioneine and the group using only calanus sinicus oil in reducing the contents of these inflammatory factors, specifically as follows: In terms of the inflammatory factor IL-6, the group using only ergothioneine was 1.38 pg / mL; the group using only calanus sinicus oil was 1.38 pg / mL; the composition group was 0.18 pg / mL; the IL-6 content of the composition group was significantly lower than that of the group using only ergothioneine and the group using only calanus sinicus oil (p < 0.01), indicating that the composition group has a significant synergistic effect in inhibiting the production of IL-6. This synergistic effect is not expected in the prior art and has been clearly confirmed by experimental data.

[0097] In terms of the inflammatory factor TNF-α, the group using only ergothioneine: 1.38 pg / mL; the group using only calanus sinicus oil: 1.38 pg / mL; the composition group: 0.18 pg / mL; similar to IL-6, the TNF-α content of the composition group was significantly lower than that of the single-use group (p < 0.01), further confirming the synergistic effect of the composition group in inhibiting the production of TNF-α. This synergistic effect is not only superior to the single-use group, but also indicates that the composition group has a significant advantage in multi-target inhibition of inflammatory reactions.

[0098] Regarding the inflammatory factor NF-κB, in the group treated with ergothioneine alone: 0.81 pg / mL; in the group treated with Temora longicornis oil alone: 0.71 pg / mL; in the composition group: 0.48 pg / mL. The content of NF-κB in the composition group was significantly lower than that in the group treated with ergothioneine alone and the group treated with Temora longicornis oil alone (p<0.01), indicating that the composition group had a significant synergistic effect in inhibiting the activity of NF-κB. NF-κB is a key signaling pathway in the inflammatory response, and its inhibition indicates that the composition group can block the progression of the inflammatory response from the source.

[0099] In Test Experimental Example 4, the anti-inflammatory effect of the composition group was significantly better than that of the single-use group, and this effect was not expected in the prior art. This synergistic effect was not only reflected in the inhibition of a single inflammatory factor, but also comprehensively inhibited the inflammatory response through a multi-target mechanism, thus providing a new solution for the treatment of cardiovascular inflammation. The discovery of this synergistic effect was not only based on rigorous experimental design and scientific statistical analysis, but also clearly confirmed by comparing experimental data.

[0100] In summary, the experimental data of Test Experimental Example 4 clearly demonstrated the synergistic effect of the composition group (ergothioneine + Temora longicornis oil) in inhibiting the inflammatory factors IL-6, TNF-α and NF-κB. This synergistic effect was not only significantly better than that of the single-use group, but also comprehensively inhibited the inflammatory response through a multi-target mechanism, providing a more efficient and stable solution for the treatment of cardiovascular inflammation. This synergistic effect provided important theoretical support for the development of highly effective anti-inflammatory dietary supplements.

[0101] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composition with anti-inflammatory function, characterized in that: include: Pegasus Daphnia Oil and Ergothioneine.

2. The composition with anti-inflammatory function as claimed in claim 1, characterized in that: In the composition, the weight ratio of the Daphnia pulex oil to ergothioneine is 100:(1-10).

3. The composition with anti-inflammatory function as claimed in claim 1, characterized in that: Also included are emulsifiers; Preferably, the emulsifier comprises beeswax or lecithin.

4. The composition with anti-inflammatory function as claimed in claim 3, characterized in that: The added amount of the emulsifier accounts for 0.5% to 1.5% of the total amount of the composition with anti-inflammatory function.

5. The composition with anti-inflammatory function as claimed in claim 1, characterized in that: The composition is an oil-in-water emulsion consisting of ergothioneine coated with nanoparticles and Daphnia pulex oil.

6. The composition with anti-inflammatory function as claimed in claim 5, characterized in that: The nanoparticles include nanoliposomes and / or polymer nanoparticles.

7. A method for preparing the composition with anti-inflammatory function as claimed in any one of claims 1 to 6, characterized in that: include: The Daphnia pulex oil and ergothioneine are mixed to obtain the composition with anti-inflammatory function.

8. The method for preparing the composition with anti-inflammatory function as claimed in claim 7, characterized in that: The method of mixing the Daphnia pulex oil and ergothioneine to obtain the composition with anti-inflammatory function comprises: Adding Pegasus daphnia oil and ergothioneine into a container and stirring and mixing to obtain a mixture; An emulsifier is added to the mixture and the mixture is stirred and mixed continuously to obtain the composition with anti-inflammatory function.

9. The method for preparing the composition having anti-inflammatory function as claimed in claim 8, characterized in that: The particles of ergothioneine are not smaller than 120 mesh; and / or, The emulsifier is beeswax or lecithin; and / or, The amount of the emulsifier added is 0.5% to 1.5% of the total amount of the composition with anti-inflammatory function.

10. Use of the composition with anti-inflammatory function as claimed in any one of claims 1 to 6 in the preparation of anti-inflammatory dietary supplement products.

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