Hematopoietic anti-aging active component extracted from fish embryo as well as extraction method and application of hematopoietic anti-aging active component

Abstract

The invention provides a hematopoietic anti-aging active component extracted from fish embryos as well as an extraction method and application of the hematopoietic anti-aging active component, and belongs to the technical field of active component extraction. The hematopoietic anti-aging active component is extracted from fish embryos which are developed from fish fertilized eggs to the earlier stage of incubation at the optimum temperature and stop development through rapid freezing, and is rich in exosomes, active protein, peptide, chondroitin sulfate, hyaluronic acid, small molecule metabolites, various active factors and the like, and all the nutritional active components are matched with one another, so that the effects of resisting aging and resisting aging are achieved. The cartilage development can be supported, the joint health is ensured, the hematopoietic effect is enhanced, and the skin aging is delayed. The orally disintegrating tablet containing the hematopoietic anti-aging active component can avoid inactivation of bioactive substances caused by high temperature, and the biological activity of the hematopoietic anti-aging active component is reserved to the maximum extent. The hematopoietic anti-aging active component and the orally disintegrating tablet are simple in preparation process and remarkable in hematopoietic anti-aging effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of active component extraction, and relates to a hematopoietic anti-aging active component extracted from fish embryos, an extraction method and an application thereof. Background Art

[0002] With the intensification of global aging, the market demand for anti-aging products continues to grow. Traditional anti-aging means such as supplementing antioxidants and peptides have problems such as low bioavailability and poor stability. Exosomes, due to naturally carrying bioactive substances such as miRNAs, proteins, and lipids, show great potential in cell-to-cell communication, tissue repair, and hematopoietic anti-aging. Currently, the main source of exosomes is mammalian cells, such as mesenchymal stem cells, but there are problems such as high extraction costs and low yields.

[0003] A fish embryo refers to the early embryo from the fertilized egg until hatching or independent feeding after cell division, tissue differentiation, and organ formation. It is rich in high-quality protein, unsaturated fatty acids, vitamins, minerals such as selenium and zinc, and rare bioactive substances such as natural astaxanthin and selenocysteine. Especially for the embryo at the pre-hatching stage, at this time, the embryo has a basic morphological structure but is not yet fully mature, and is in the transition stage from "structure construction" to "function improvement". It is a critical window period for exosome secretion, antioxidant system activation, and organ primordium formation. Many exosomes produced at this time are not present in ordinary fish eggs at all and can only be formed during embryonic development. They are rich in various types and high concentrations of active substances and participate in tissue coordinated development and organ formation. In addition, cartilage is formed but not hardened during this period, and may contain active factors that promote osteogenic differentiation. The cartilage is more nutritious and easier to absorb after enzymatic hydrolysis with collagenase, and high-quality type II collagen peptides, chondroitin sulfate, hyaluronic acid and other nutrients can be obtained, and active substances such as TGF-β, IGF-1, and BMP-2 are released, which has the function of promoting cartilage regeneration and differentiation.

[0004] Currently, there is no research report on extracting a hematopoietic anti-aging active component rich in exosomes from fish embryos and its application. Summary of the Invention

[0005] Based on the above purpose, the present invention provides a hematopoietic anti-aging active component extracted from fish embryos, an extraction method and an application thereof.

[0006] To achieve the above purpose, the present invention adopts the following technical solutions: In the first aspect, the present application provides a hematopoietic anti-aging active component extracted from fish embryos, and the extraction method of this active component includes: S01: Wash the fish embryos that have developed to the pre-hatching stage after fertilization with ice-cold PBS, add pre-cooled PBS, and homogenize to obtain a homogenate.

[0007] Since temperature is a key factor affecting the development rate of fish embryos, in this invention, fish embryos are all taken from those that have developed to the pre-hatching stage at their optimal hatching temperature and then rapidly frozen to stop development. After gently washing the fish eggs twice with ice-cold PBS, the fish embryos and pre-cooled PBS at 0 - 8°C are mixed at a mass-to-volume ratio of 1 g : 1 - 3 mL, and gently homogenized 2 - 3 times for 5 - 10 s each time to obtain a homogenate. More preferably, the mass-to-volume ratio of fish embryos to pre-cooled PBS is 1 g : 2 mL. Washing with ice-cold PBS can reduce the temperature of the fish embryos, reduce their metabolic activities, and extend their storage time. In addition, mixing with pre-cooled PBS can keep the fish embryos within an appropriate pH range, thus maintaining the biological activity of the fish embryos, further maintaining their integrity, and preventing mildew.

[0008] In this application, fish embryos are selected to extract hematopoietic anti-aging active components containing exosomes, especially fish embryos that have developed to the pre-hatching stage after fertilization. Fish embryos at this stage are in the critical period of rapid development. The secretion amount of exosomes and the production and release amounts of various enzymes both reach their peaks. Therefore, the hematopoietic anti-aging active components prepared from pre-hatching fish embryos are rich in exosomes with high amounts and high activities of substances, very rich active proteins, peptides, chondroitin sulfate, hyaluronic acid, and small molecule metabolites, etc. These various nutritional active components cooperate with each other to enhance the hematopoietic anti-aging effect and can more effectively exert the hematopoietic anti-aging efficacy.

[0009] S02: After the homogenate is mixed with collagenase II, it is enzymatically digested at room temperature for 2 - 4 h, and the enzymatic digestion product is filtered to obtain the hematopoietic anti-aging active component.

[0010] The homogenate is mixed with collagenase II and enzymatically digested at room temperature for 2 - 4 h to obtain an enzymatic digestion product. Among them, the mass-to-volume of collagenase II to the homogenate is 0.1 - 0.3% g / mL. The enzymatic digestion product is filtered successively through nylon mesh cloths with 40 meshes, 100 meshes, and 200 meshes to remove the residue, and the hematopoietic anti-aging active component extracted from fish embryos is obtained. More preferably, the mass-to-volume of collagenase II to the homogenate is 0.2% g / mL, and the enzymatic digestion time is 3 h.

[0011] Fish embryos that have developed to the pre-hatching stage after fertilization have produced cartilage but have not formed hard bones. At this time, to enzymatically digest and differentiate the cartilage, collagenase II is used in this application to enzymatically digest the homogenate formed by fish embryos. After the cartilage is enzymatically digested by collagenase II, high-quality type II collagen peptides, chondroitin sulfate, hyaluronic acid, and other nutrients can be obtained, and at the same time, active substances such as TGF-β, IGF-1, and BMP-2 are released. This makes the enzymatic digestion product have the effect of promoting cartilage regeneration and differentiation. After the enzymatic digestion product is filtered to retain the filtrate and remove the residue, the above-mentioned nutrients and active substances can be retained to form the hematopoietic anti-aging active component.

[0012] In a second aspect, the hematopoietic anti-aging active component extracted from fish embryos provided in the present application is used for preparing a drug or health product with hematopoietic anti-aging efficacy, and the drug or health product is an orally disintegrating tablet.

[0013] In a third aspect, the present application provides an orally disintegrating tablet, which includes the hematopoietic anti-aging active component in the first aspect, D-mannitol, ginger extract, peppermint powder and gelatin. Among them, D-mannitol is a filler, ginger extract and peppermint powder are flavoring agents, and gelatin is a thickening agent. Before use, gelatin needs to be dissolved in water at 65°C in advance. The preparation method of the orally disintegrating tablet is prepared according to the existing preparation method, such as stirring and mixing components, degassing, injecting into a mold, pre-freezing, sublimation drying, analytical drying by freeze-drying and heat sealing.

[0014] Preferably, based on the hematopoietic anti-aging active component, the addition amounts of D-mannitol, ginger extract, peppermint powder and gelatin are 0 - 10% g / mL, 0 - 1.0% g / mL, 0 - 0.5% g / mL and 0 - 2% g / mL respectively.

[0015] More preferably, based on the hematopoietic anti-aging active component, the addition amounts of D-mannitol, ginger extract, peppermint powder and gelatin are 1% g / mL, 0.6% g / mL, 0.3% g / mL and 0.5% g / mL respectively.

[0016] The present invention has the following beneficial effects: (1) The present application extracts for the first time a hematopoietic anti-aging active component containing exosomes from fish embryos, especially fish embryos at the pre-hatching stage. Fish embryos at this stage are in the critical period of rapid development. The secretion amount of exosomes and the production and release amounts of various enzymes reach their peaks. Therefore, the hematopoietic anti-aging active component prepared from fish embryos at the pre-hatching stage is rich in exosomes with high amounts and high activities of substances, very rich active proteins, peptides, chondroitin sulfate, hyaluronic acid and small molecule metabolites, etc. Each nutritional active ingredient cooperates with each other to enhance the hematopoietic anti-aging effect and can more effectively exert the hematopoietic anti-aging efficacy.

[0017] (2) Fish embryos at the pre-hatching stage after fertilization are also the critical period of cartilage development. By using collagenase to enzymatically hydrolyze fish embryos at this stage in the present application, the enzymatic hydrolysis products can contain active factors such as TGF-β, IGF-1, BMP-2 that promote osteogenic differentiation, as well as nutrients such as type II collagen peptides, chondroitin sulfate, and hyaluronic acid. Furthermore, the hematopoietic anti-aging active component can support cartilage development, ensure joint health and delay skin aging.

[0018] (3) The hematopoietic anti-aging active component extracted from fish embryos in the orally disintegrating tablet in the present application. The combination of the two can avoid the inactivation of bioactive substances caused by high temperature and maximally retain the biological activity of the hematopoietic anti-aging component.

[0019] (4) The orally disintegrating tablets in this application can rapidly disintegrate in the oral cavity, significantly improving the oral absorption efficiency of exosomes and the utilization rate of highly bioactive substances, effectively exerting the hematopoietic anti-aging effect, being applicable to uses such as promoting hematopoiesis, delaying aging, and improving skin aging, and having both convenience and stability.

[0020] (5) The preparation process of the hematopoietic anti-aging active component and the orally disintegrating tablets in this application is simple, and the hematopoietic anti-aging effect is remarkable. Description of the Drawings

[0021] Figure 1 TEM image and distribution diagram of the size and number of exosome vesicles after centrifugation of the hematopoietic anti-aging active component prepared in Example 1; among them, A is the TEM image, and B is the distribution diagram of the size and number of exosome vesicles; Figure 2 Appearance diagram of the orally disintegrating tablets prepared in Example 4; Figure 3 Diagram of the influence of the orally disintegrating tablets prepared in Example 4 at different concentrations on the colony morphology and the number of colony CFUs of hematopoietic stem cells. Among them, A is the colony morphology diagram of hematopoietic stem cells in the blank group, B is the colony morphology diagram of hematopoietic stem cells in the low-dose group, and C is the colony morphology diagram of hematopoietic stem cells in the high-dose group; D is the diagram of the influence of the orally disintegrating tablets on the number of colony CFUs of hematopoietic stem cells, ** significantly different from the blank group (P<0.01); Figure 4 Diagram of the escape latency of mice in each group during the positioning navigation test, where * is significantly different from the normal group (P<0.05), and # is significantly different from the model group (P<0.05); Figure 5 Diagram of the proportion of the residence time of mice in each group in the original platform quadrant during the spatial exploration test, where ** is significantly different from the normal group (P<0.01), and # is significantly different from the model group (P<0.05); Figure 6 Diagram of the muscle strength test results of mice in each group, where * is significantly different from the normal group (P<0.05); ** is significantly different from the normal group (P<0.01); Figure 7 Diagram of the anti-fatigue ability test results of mice in each group; among them, ** is significantly different from the normal group (P<0.01), and ## is significantly different from the model group (P<0.01); Figure 8 Diagram of the TNF-α concentration in the serum of mice in each group, where * is significantly different from the normal group (P<0.05); Figure 9IL-1β concentrations in the sera of mice in each group, where *, significantly different from the normal group (P < 0.05), **, significantly different from the normal group (P < 0.01); #, significantly different from the model group (P < 0.05); Figure 10 T-AOC concentrations in the sera of mice in each group, where **, significantly different from the normal group (P < 0.01); ##, significantly different from the model group (P < 0.01); Figure 11 Comparison diagrams of skin elasticity detection of mice in each group, where *, significantly different from the normal group (P < 0.05); **, significantly different from the normal group (P < 0.01), #, significantly different from the model group (P < 0.05); Figure 12 Comparison diagrams of osteophyte volume of the bone joints of mice in each group, where **, significantly different from the normal group (P < 0.01); ##, significantly different from the model group (P < 0.01). Specific implementation manners

[0022] The technical solutions of the present invention will be further explained and illustrated by specific embodiments below.

[0023] Example 1 The embodiment of the present application provides a hematopoietic anti-aging active component extracted from fish embryos. The extraction method of the active component includes: S101: Take turbot fish embryos that have been fertilized and developed at the optimal hatching temperature of 19 - 21°C for 48 h and then rapidly frozen to stop development. Gently wash the fish embryos 2 times with ice-cold PBS. Mix the fish eggs and PBS pre-cooled to 0°C at a mass-to-volume ratio of 1 g:2 mL, and gently homogenize 3 times, 5 s each time, to obtain a homogenate.

[0024] S102: Mix the homogenate with collagenase II and enzymatically digest at room temperature for 3 h to obtain an enzymatic digestion product. Among them, the mass-to-volume ratio of collagenase II to the homogenate volume is 0.2% g / mL. The enzymatic digestion product is filtered through nylon mesh cloths with 40 meshes, 100 meshes, and 200 meshes in sequence to remove residues, and a hematopoietic anti-aging active component extracted from fish embryos is obtained.

[0025] Example 2 The embodiment of the present application provides a hematopoietic anti-aging active component extracted from fish embryos. The extraction method of the active component includes: S201: Take turbot fish embryos that have been fertilized and developed at the optimal hatching temperature of 13 - 15°C for 108 h and then rapidly frozen to stop development. Gently wash the fish embryos 2 times with ice-cold PBS. Mix the fish embryos and PBS pre-cooled to 5°C at a mass-to-volume ratio of 1 g:1 mL, and gently homogenize 2 times, 10 s each time, to obtain a homogenate.

[0026] S202: Mix the homogenate with collagenase II and enzymatically digest at room temperature for 4 h to obtain an enzymatically digested product. Among them, the mass-to-volume ratio of collagenase II to the volume of the homogenate is 0.3% g / mL. The enzymatically digested product is sequentially filtered through nylon mesh cloths with 40 meshes, 100 meshes, and 200 meshes to remove residues, thereby obtaining the hematopoietic anti-aging active component extracted from fish embryos.

[0027] Example 3 The embodiment of the present application provides a hematopoietic anti-aging active component extracted from fish embryos. The extraction method of this active component includes: S301: Take grouper fish embryos that have been fertilized and developed for 22 h at the optimal hatching temperature of 26 °C and rapidly frozen to stop development, and gently wash the fish embryos 2 times with ice-cold PBS. Mix the fish embryos and PBS pre-cooled to 8 °C at a mass-to-volume ratio of 1 g: 3 mL, and gently homogenize 3 times, 8 s each time, to obtain a homogenate.

[0028] S302: Mix the homogenate with collagenase II and enzymatically digest at room temperature for 2 h to obtain an enzymatically digested product. Among them, the mass-to-volume ratio of collagenase II to the volume of the homogenate is 0.1% g / mL. The enzymatically digested product is sequentially filtered through nylon mesh cloths with 40 meshes, 100 meshes, and 200 meshes to remove residues, thereby obtaining the hematopoietic anti-aging active component extracted from fish embryos.

[0029] Example 4 The embodiment of the present application provides an orally disintegrating tablet, which includes the hematopoietic anti-aging active component in Example 1, D-mannitol, ginger extract, peppermint powder, and gelatin. Among them, based on the added volume of the hematopoietic anti-aging active component in Example 1, the addition amounts of D-mannitol, ginger extract, peppermint powder, and gelatin are 1% g / mL, 0.6% g / mL, 0.3% g / mL, and 0.5% g / mL, respectively.

[0030] Example 5 The embodiment of the present application provides an orally disintegrating tablet, which includes the hematopoietic anti-aging active component in Example 2, D-mannitol, ginger extract, peppermint powder, and gelatin. Among them, based on the added volume of the hematopoietic anti-aging active component in Example 2, the addition amounts of D-mannitol, ginger extract, peppermint powder, and gelatin are 0 - 10% g / mL, 0 - 1.0% g / mL, 0 - 0.5% g / mL, and 0 - 2% g / mL, respectively.

[0031] Example 6 An embodiment of the present application provides an orally disintegrating tablet, which includes the hematopoietic anti-aging active component in Example 3, D-mannitol, ginger extract, mint powder and gelatin. Among them, based on the added volume of the hematopoietic anti-aging active component in Example 3, the addition amounts of D-mannitol, ginger extract, mint powder and gelatin are 0-10% g / mL, 0-1.0% g / mL, 0-0.5% g / mL and 0-2% g / mL respectively.

[0032] To verify that the hematopoietic anti-aging active component extracted from fish embryos contains exosomes, in this application, the hematopoietic anti-aging active component prepared in Example 1 is taken as an example for verification tests. The specific experimental process is as follows: At 4°C, the hematopoietic anti-aging active component prepared in Example 1 is centrifuged at 300g, 2000g, 20000 g, and 100000g for 15 minutes, 15 minutes, 30 minutes, and 70 minutes respectively; after centrifugation, the precipitate is resuspended in ice-cold PBS, filtered through a 0.22 μm pore size filter membrane, and stored temporarily at -80°C. Transmission electron microscopy is used to observe the structure of the precipitate, and a nanoparticle tracking analyzer is used to detect the size and number distribution map of exosome vesicles in the precipitate, obtaining Figure 1 .

[0033] It can be seen from Figure 1 that there are many complete oval membrane-like structure vesicles in the centrifuged component of the hematopoietic anti-aging active component prepared in Example 1; at the same time, it is detected by a nanoparticle tracking analyzer that the peak diameter of the vesicles is 127.4 nm, and the number of vesicles is 3.5×10 6 per mL, and the proportion of vesicles with a diameter of 127.4±71.3 nm is 97.6%, which conforms to the typical characteristics of exosomes, indicating that the hematopoietic anti-aging active component prepared in Example 1 contains exosomes.

[0034] This application also conducts appearance quality inspection, hematopoietic efficacy inspection and anti-aging efficacy inspection on the orally disintegrating tablets prepared in Example 4 respectively. The specific contents are as follows: 1. Appearance quality inspection The orally disintegrating tablets prepared in Example 4 are respectively inspected for appearance, taste, disintegration time limit and redispersibility indexes, and Table 1 and Figure 2 .

[0035] Table 1: Appearance quality inspection results of orally disintegrating tablets It can be seen from Table 1 and Figure 2 that the orally disintegrating tablets prepared in Example 4 are light pink, porous, regular in shape, uniform in color, plump, non-sticking to the wall, and the disintegration time is 25 s, with good quality.

[0036] 2. Hematopoietic efficacy inspection Disperse the orally disintegrating tablets prepared in Example 4 in water and dissolve them. After filtering through a coarse filter paper and a 0.22 μm filter membrane, dilute the filtered stock solution by 10 times and 100 times respectively. Isolate and extract umbilical cord blood mononuclear cells (MNCs) according to the existing conventional method. Add MNCs suspension with a final concentration of 5×10 3 / mL and a volume of 250 μL to 5 mL of MethoCult™ H4434 Classic medium. At the same time, add 250 μL of PBS buffer solution, the filtered stock solution, and the filtered stock solution diluted 10 times respectively to form a blank group, a high-dose group, and a low-dose group. Mix well and inoculate into a 6-well plate, and place 1.1 mL of the mixed solution in each well. Place the 6-well plate in an incubator at 37°C and 5% CO2 for 14 days, observe the colony morphology, take pictures, and count to obtain the attached Figure 3 .

[0037] As can be seen from the attached Figure 3 Most of the colonies in each group are erythroid burst-forming unit (BFU-E) colonies with a red color, and there are almost no other colony types. Compared with the blank group, the colony volume of hematopoietic stem cells in the high-dose group and the low-dose group is larger, and the number of colonies (CFU) in the high-dose group is significantly increased (P<0.05), indicating that the orally disintegrating tablets prepared in Example 4 have the function of promoting hematopoiesis in vitro.

[0038] 3. Anti-aging efficacy detection In this application, a mouse aging model is established and the orally disintegrating tablets prepared in Example 4 are administered by gavage to observe the changes in anti-aging indicators and verify its anti-aging efficacy.

[0039] Specifically, select 24 SPF-grade female KM mice with a body weight of 18 - 22 g. After 1 week of adaptive feeding, divide them into 3 groups: a normal group, a model group, and a Japanese flounder embryo group. Each group of mice is administered at a volume of 0.1 mL / 10 g BW, weighed once a day, and the dosage is adjusted according to the body weight. The feeding cycle is 8 weeks.

[0040] Normal group: Inject 0.9% normal saline subcutaneously at the same site on the back of the neck of the mice every day; Model group: Inject 200 mg / kg of D-galactose solution subcutaneously at the same site on the back of the neck of the mice every day; Japanese flounder embryo group: Inject 200 mg / kg of D-galactose solution subcutaneously at the same site on the back of the neck of the mice every day, and at the same time administer the orally disintegrating tablets prepared in Example 4 by gavage at 20 mg / kg.

[0041] After the feeding ended, the anti-aging function of the orally disintegrating tablets prepared in Example 4 was verified from the senescence signs, spatial learning and memory ability, muscle strength, anti-fatigue ability, in vivo inflammation level, body antioxidant ability, skin elasticity, and osteophyte volume of the mice in each group.

[0042] (1) Senescence signs The hair of the mice in the normal group was smooth, shiny, and had good skin elasticity. They had good mental state, loved to move, and had normal diet, drinking water, defecation, and urination. The hair of the mice in the model group was somewhat sparse, dull, and easy to fall off. They were listless, with significantly reduced activity, increased food intake, and decreased frequency of defecation and urination. The signs of the mice in the Japanese flounder embryo group were similar to those in the normal group, with almost no difference.

[0043] (2) Spatial learning and memory ability of mice In the examples of this application, the Morris water maze experiment was used to evaluate the spatial learning and memory ability of mice. Among them, the test content of the Morris water maze experiment included the place navigation test and the spatial exploration experiment. Its specific steps were carried out according to the existing method, and the attached Figure 4 、 5 .

[0044] As can be seen from the attached Figure 4 , in the place navigation test, as the number of training days increased, the escape latency of the mice in the normal group and the Japanese flounder embryo group gradually shortened; while the escape latency of the mice in the model group had little relationship with the number of training days; especially in the final place navigation, the escape latency of the Japanese flounder embryo group was significantly lower than that of the model group (P<0.05), and there was no significant difference from the normal group.

[0045] As can be seen from the attached Figure 5 , in the spatial exploration test, compared with the normal group, the time that the mice in the model group stayed in the original platform quadrant was significantly reduced, while there was no significant difference in the Japanese flounder embryo group; compared with the model group, the time that the mice in the Japanese flounder embryo group stayed in the original platform increased significantly.

[0046] The above results show that the spatial learning ability and memory ability of D-galactose-induced senescent mice are impaired, while the orally disintegrating tablets prepared in Example 4 can improve the spatial learning ability and memory ability of D-galactose-induced senescent mice.

[0047] (3) Muscle strength A four-limb tensiometer was used to test the muscle strength of the mice in each group, and the test results are as Figure 6 shown. As can be seen from the attached Figure 6 , compared with the normal group, the four-limb grasping force of the mice in the model group was significantly reduced; the four-limb grasping force of the mice in the Japanese flounder embryo group was between that of the normal group and the model group, and the four-limb grasping force was increased compared with the model group. This indicates that the orally disintegrating tablets prepared in Example 4 can increase the muscle strength of D-galactose-induced senescent mice.

[0048] (4)Anti-fatigue ability The anti-fatigue ability of each group of mice was tested using a rotary fatigue tester, and the number of drops of each group of mice was recorded to obtain the appendix Figure 7 . From the appendix Figure 7 It can be seen that compared with the normal group, the number of drops of the mice in the model group increased significantly, while there was no significant difference in the flounder embryo group; compared with the model group, the number of drops of the mice in the flounder embryo group decreased significantly. This indicates that the orally disintegrating tablets prepared in Example 4 can enhance the motor flexibility and endurance of D-galactose-induced aging mice.

[0049] (5)Inflammatory level in vivo The aging process is often accompanied by the occurrence of inflammatory reactions, and chronic inflammation will further exacerbate the aging of the body. As an inflammatory factor, TNF-α can regulate immune function and mediate the occurrence of inflammatory reactions; IL-1β can damage the extracellular fluid, change the permeability of blood vessels, and damage the body. Therefore, TNF-α and IL-1β in serum can objectively and accurately judge the degree of inflammation of the body. In the examples of this application, the blood of each group of mice was taken respectively, the serum in the blood was extracted, and the concentrations of TNF-α and IL-1β in the serum were detected to obtain the appendix Figure 8 , 9 .

[0050] From the appendix Figure 8 It can be seen that compared with the normal group, the serum TNF-α concentration of the mice in the model group increased significantly, while there was no significant difference in the flounder embryo group. From the appendix Figure 9 It can be seen that compared with the normal group, the serum IL-1β concentration of the mice in the model group increased significantly, while the flounder embryo group was significantly lower than the model group. The above indicates that the orally disintegrating tablets prepared in Example 4 can reduce the level of inflammatory factors in vivo, and thus delay the aging of the body.

[0051] (6)Antioxidant capacity of the body T-AOC (Total antioxidant capacity) is an important index to evaluate the overall efficiency of the body's antioxidant defense system, reflecting the comprehensive ability of all antioxidant substances in the body to synergistically neutralize free radicals and resist oxidative stress. When the T-AOC level decreases, it indicates insufficient antioxidant reserve, which may be related to aging and inflammation. In the examples of this application, the blood of each group of mice was taken respectively, the serum in the blood was extracted, and the concentration of T-AOC in the serum was detected to obtain the appendix Figure 10 .

[0052] From the appendix Figure 10 It can be known that compared with the normal group, the serum T-AOC concentration of the mice in the model group decreased significantly, while the flounder embryo group was significantly higher than the model group. This indicates that the orally disintegrating tablets prepared in Example 4 can enhance the antioxidant capacity of the body, and thus delay the aging of the body.

[0053] (7)Skin elasticity There is a close relationship between aging and skin elasticity. Aging is often accompanied by the loss of skin collagen, the degradation of elastin, the accumulation of free radicals that damage cells, etc., resulting in a decrease in skin elasticity. In the embodiments of the present application, a skin elasticity tester MPA580 was used to test the skin elasticity of each group of mice, and the attached Figure 11 .

[0054] As can be seen from the attached Figure 11 , compared with the normal group, the R2 value of the skin elasticity of the mice in the model group was significantly reduced, while the R2 value of the skin elasticity of the mice in the Japanese flounder embryo group was significantly increased compared with the model group. This indicates that the orally disintegrating tablets prepared in Example 4 can delay skin aging and increase skin elasticity.

[0055] (8)Osteophyte volume Osteophytes are abnormal bony protrusions at the joint margins, formed by endochondral ossification, and are common in aging joints. Osteophyte volume is a core indicator for evaluating bone aging. In the embodiments of the present application, Micro-CT was used to accurately measure and quantitatively analyze the osteophyte volume of the bone joints of each group of mice, and the attached Figure 12 .

[0056] As can be seen from the attached Figure 12 , compared with the normal group, the osteophyte volume of the bone joints of the mice in the model group was significantly increased, while the osteophyte volume of the mice in the Japanese flounder embryo group was almost indistinguishable from that of the normal group. This indicates that the orally disintegrating tablets prepared in Example 4 can delay bone joint aging and maintain the health of bone joints.

[0057] In summary, it can be seen that the orally disintegrating tablets containing hematopoietic anti-aging active components prepared in Example 4 of the present application do have the functions of delaying the aging signs of mice, delaying organ aging, improving the spatial learning and memory ability of aging mice, increasing muscle strength, enhancing motor flexibility and endurance, reducing inflammation in the body, enhancing the antioxidant capacity of the body, and delaying skin aging.

[0058] The above are only the preferred embodiments of the present application and are not used to limit the present application. For those skilled in the art, the present application can have various changes and modifications. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for extracting a hematopoietic anti-aging active component extracted from fish embryos, characterized in that, Comprising: Washing fish embryos that have developed to the pre-hatching stage after fertilization with ice-cold PBS, adding pre-chilled PBS, and homogenizing to obtain a homogenate. After mixing the homogenate with collagenase II, enzymatically digest at room temperature for 2 - 4 h, and sequentially filter the enzymatic digestion products through nylon mesh cloth with 40 meshes, 100 meshes, and 200 meshes to obtain the hematopoietic anti-aging active component.

2. The extraction method of the hematopoietic anti-aging active component extracted from fish embryos according to claim 1, characterized in that, The mass-volume ratio of the fish embryos to the pre-chilled PBS is 1 g: 1 - 3 mL.

3. The extraction method of the hematopoietic anti-aging active component extracted from fish embryos according to claim 1, wherein The mass ratio of the added mass of collagenase II to the volume of the homogenate is 0.1 - 0.3% g / mL.

4. A hematopoietic anti-aging active component extracted from fish embryos, characterized in that, Obtained by the extraction method according to any one of claims 1 - 3.

5. The hematopoietic anti-aging active component obtained by the extraction method according to any one of claims 1 - 3 is used for preparing a drug or health product with hematopoietic anti-aging efficacy.

6. The application according to claim 5, characterized in that, The drug or health product is an orally disintegrating tablet.

7. An orally disintegrating tablet, characterized in that, Comprising the hematopoietic anti-aging active component obtained by the extraction method according to any one of claims 1 - 3, D-mannitol, ginger extract, peppermint powder, and gelatin.

8. The orally disintegrating tablet according to claim 7, characterized in that, Based on the hematopoietic anti-aging active component, the addition amounts of D-mannitol, ginger extract, peppermint powder, and gelatin are respectively 0 - 10% g / mL, 0 - 1.0% g / mL, 0 - 0.5% g / mL, and 0 - 2% g / mL.

9. The orally disintegrating tablet according to claim 7, wherein Based on the hematopoietic anti-aging active component, the addition amounts of D-mannitol, ginger extract, peppermint powder, and gelatin are respectively 1% g / mL, 0.6% g / mL, 0.3% g / mL, and 0.5% g / mL.

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