Nano-particle, preparation method thereof and application of nano-particle in anti-aging
By preparing nanoparticles composed of Chinese herbal extracellular vesicles, the problems of poor blood circulation and oxidative stress in skin aging were solved, achieving significant anti-aging effects and improved nematode survival rates.
Patent Information
- Application Number
- CN202510928720.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-07
- Publication Date
- 2025-09-26
AI Technical Summary
Existing technologies are difficult to effectively alleviate skin aging, especially by improving aging problems caused by poor circulation of Qi and blood and oxidative stress.
Nanoparticles composed of Chinese herbal extracellular vesicles were used to prepare Astragalus and Salvia miltiorrhiza nanoparticles (ASNPs), taking advantage of their anti-oxidative stress and blood-activating and qi-invigorating properties. The preparation method includes juicing, centrifugation and purification steps of Astragalus and Salvia miltiorrhiza stock solution to form stable nanoparticles.
It significantly alleviates cell aging, improves the survival rate of nematodes, and resists the oxidative stress of endogenous superoxide radical generators in cells, providing a new idea for anti-aging and anti-oxidative stress.
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Figure CN120695066A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of biomedicine technology, and in particular to a nanoparticle, a preparation method thereof, and an application thereof in anti-aging. Background Art
[0002] As the largest organ in the human body, the skin is also the first line of defense against the external environment. It reflects various physiological and pathological changes in the human body, including the aging process. Age-related changes in the skin are closely related to overall aging. The Yellow Emperor's Classic of Internal Medicine discusses the aging process. For example, the "Suwen: On the Original Nature of Ancient Times" states: "At seven or eight... the essence of heaven is exhausted, the essence is scarce, the kidneys weaken, and the body reaches its limit. At eight or eight, the teeth and hair fall out." It also points out that the facial skin is an outward manifestation of the changes in qi, blood, yin and yang within the internal organs. For example, the "Lingshu: On the Forms of Evil Qi and Organ Diseases" states: "The meeting point of all yang is the face... The twelve meridians and three hundred and sixty-five collaterals all flow upwards to the face and flow through the orifices..." The "Suwen: On the Six Sections of Organs and Images" states: "The heart, its brilliance is in the face." And the "Suwen: On the Creation of the Five Zang Organs" states: "The lungs are connected to the skin and their glory is the hair." These descriptions correspond to different parts of the face and organs.
[0003] Traditional Chinese Medicine (TCM) posits that the signs of skin aging, such as dullness, wrinkles, and sagging, are primarily due to problems with Qi and blood. Qi is the leader of blood, and blood is the mother of Qi. Qi and blood are interdependent. When Qi is deficient, the force driving blood circulation is insufficient, leading to blood deficiency; blood deficiency, in turn, impairs the production of Qi. The two complement each other, jointly maintaining vital activities and a radiant complexion. The aging process of the skin largely reflects Qi and blood deficiency, or poor circulation. Activating blood circulation and replenishing Qi are crucial treatments that target this core pathological link.
[0004] From an evolutionary and ecological perspective, the defensive substances produced by medicinal plants in response to adversity, in addition to improving their own adaptation to environmental stress, can also produce good biocompatibility with the human body and be used to treat human diseases. This evolutionary law determines the multi-component material basis of traditional Chinese medicine. Due to their closed structure, extracellular vesicles derived from traditional Chinese medicine often contain a large amount of intracellular bioactive substances such as proteins and nucleic acids, and are one of the main material bases for intercellular communication. Studies on extracellular vesicles of Chinese herbal medicines have found that different nanovesicles can be effectively taken up by different cells, and deliver substances into cells or further change cell functions. Therefore, the present invention specifically proposes a nanoparticle composed of extracellular vesicles of Chinese herbal medicines. Summary of the Invention
[0005] The purpose of the present invention is to provide a nanoparticle and a preparation method thereof and application in anti-aging to solve the problems existing in the above-mentioned prior art. The ASNPs prepared by the present invention can significantly alleviate cell aging; resist oxidative stress induced by endogenous superoxide free radical generator PQ in cells, and improve the survival rate of nematodes, providing a new idea and technical basis for alleviating aging and resisting oxidative stress.
[0006] To achieve the above object, the present invention provides the following solutions:
[0007] The present invention provides a method for preparing nanoparticles, comprising the following steps:
[0008] After adding PBS, astragalus and salvia miltiorrhiza were squeezed and filtered to obtain astragalus and salvia miltiorrhiza stock solution;
[0009] The Astragalus and Salvia miltiorrhiza stock solution was centrifuged six times to obtain a nanoparticle solution;
[0010] The nanoparticle solution is purified to obtain nanoparticles.
[0011] Optionally, the six centrifugation operations are:
[0012] Centrifuging the Astragalus and Salvia miltiorrhiza stock solution at 1000×g for 15 minutes at 4° C. to obtain a first centrifuge solution;
[0013] Centrifuge the supernatant of the first centrifuge again at 3000×g for 30 min at 4°C to obtain a second centrifuge;
[0014] The supernatant of the second centrifuge was centrifuged again at 10,000 × g for 30 min at 4°C to obtain a third centrifuge;
[0015] The supernatant of the third centrifuge was centrifuged again at 10,000 × g for 10 min at 4° C. to obtain a fourth centrifuge;
[0016] Filtering the supernatant of the fourth centrifuge through a 0.45 μm filter membrane and a 0.22 μm filter membrane to obtain a filtrate;
[0017] The filtrate was ultracentrifuged at 100,000×g for 90 minutes at 4° C. to obtain a fifth centrifuge;
[0018] The precipitate of the fifth centrifuge was resuspended, and ultracentrifuged at 100,000 × g for 90 min at 4° C. to obtain a sixth centrifuge;
[0019] The precipitate of the sixth centrifugation liquid is resuspended to obtain a nanoparticle solution.
[0020] Optionally, the mass ratio of the astragalus root to the salvia miltiorrhiza root is 1.5:1-2.5:1.
[0021] Optionally, the juice squeezing operation is performed by squeezing for 1 minute, stopping for 2 minutes, and squeezing 7 times in total.
[0022] Optionally, the purification operation is:
[0023] adding the nanoparticle solution to a sucrose density gradient solution and centrifuging; the sucrose density gradient solution is 8%, 15%, 30%, 45% and 60% sucrose solution;
[0024] The nanoparticles in the 30% sucrose solution and 45% sucrose solution layers were aspirated, and resuspended by centrifugation to obtain ASNPs nanoparticles.
[0025] Optionally, the centrifugation conditions are 150,000 g ultracentrifugation for 120 min at 4°C.
[0026] The present invention also provides nanoparticles obtained according to the preparation method.
[0027] The present invention also provides use of the nanoparticles in preparing anti-aging products.
[0028] The present invention also provides an anti-aging product comprising the nanoparticles.
[0029] Optionally, pharmaceutically acceptable excipients are also included.
[0030] The present invention discloses the following technical effects:
[0031] Through a systematic analysis of 12,583 prescriptions in the "Compendium of Traditional Chinese Medicine Formulas," the present invention discovered that the Astragalus and Salvia miltiorrhiza combination appears in 17.4% of prescriptions, far exceeding other Qi-invigorating and blood-activating herbal combinations, making it a classic Qi-invigorating and blood-activating herbal combination. Therefore, the present invention selected Astragalus and Salvia miltiorrhiza as raw materials to extract their exosome vesicles and prepare nanoparticles.
[0032] The present invention provides a method for extracting and preparing ASNPs nanoparticles, which can successfully extract Astragalus and Salvia miltiorrhiza nanoparticles (ASNPs). Cell and animal experiments have demonstrated that the ASNPs prepared by the present invention can significantly alleviate cellular aging, resist oxidative stress induced by the endogenous superoxide radical generator PQ, and improve the survival rate of nematodes. This invention provides a new approach and technical foundation for alleviating aging and resisting oxidative stress, and the ASNPs prepared by the present invention have broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0033] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.
[0034] Figure 1 The electron micrographs of ASNPs prepared by the conventional method after freeze-drying and the extraction process of the present invention are shown;
[0035] Figure 2 Transmission electron microscopy image of ASNPs prepared in the present invention;
[0036] Figure 3 The average particle size analysis results of ASNPs prepared in the present invention; the left figure shows: Astragalus-derived nanoparticles (ASNP-A); the right figure shows: Salvia-miltiorrhiza-derived nanoparticles (ASNP-S);
[0037] Figure 4 The average potential analysis results of ASNPs prepared in the present invention; the left figure shows: Astragalus-derived nanoparticles (ASNP-A); the right figure shows: Salvia-miltiorrhiza-derived nanoparticles (ASNP-S);
[0038] Figure 5 The staining results of the ASNPs prepared by the present invention on the human vascular endothelial cell aging model and the positive percentage of β-galactosidase staining;
[0039] Figure 6 The staining results of the ASNPs prepared in the present invention on the fibroblast aging model and the positive percentage of β-galactosidase staining;
[0040] Figure 7 The ASNPs prepared in the present invention affect the transcription of the type I collagen encoding gene Col1a1 in mouse fibroblasts;
[0041] Figure 8 This is the effect of the ASNPs prepared in the present invention on the survival rate of nematodes. DETAILED DESCRIPTION
[0042] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.
[0043] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. The intermediate value within any stated value or stated range, and each smaller range between any other stated value or intermediate value within the stated range, is also encompassed within the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.
[0044] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.
[0045] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be exemplary only.
[0046] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.
[0047] Example 1
[0048] 1. Preparation of ASNPs stock solution
[0049] Fresh astragalus and salvia miltiorrhiza were weighed, washed and peeled, and then washed together with instruments with pure water. The washed astragalus and salvia miltiorrhiza were cut into small pieces as much as possible using the washed instruments. A certain amount of pre-cooled PBS (volume ratio of 1:3) was added to the cut small pieces of astragalus and salvia miltiorrhiza (weight ratio of 1.5:1-2.5:1, and 1.5:1 was used in this embodiment) and then juiced. The juice was squeezed for 1 minute and stopped for 2 minutes for a total of 7 times to obtain a squeezed juice. The squeezed juice was filtered using sterile gauze to obtain an astragalus and salvia miltiorrhiza stock solution.
[0050] 2. Preparation of ASNPs Nanoparticles
[0051] 1) Remove the Astragalus and Salvia miltiorrhiza stock solution and transfer it to a first centrifuge tube. Centrifuge at 1000 × g for 15 min at 4°C to obtain a first centrifuge solution.
[0052] 2) Transfer the supernatant of the first centrifuge to a second centrifuge tube and centrifuge again at 3000 × g for 30 min at 4°C to obtain a second centrifuge.
[0053] 3) Transfer the supernatant of the second centrifuge to a third centrifuge tube and centrifuge again at 10,000 × g for 30 min at 4°C to obtain a third centrifuge;
[0054] 4) Transfer the supernatant of the third centrifuge to a fourth centrifuge tube and centrifuge again at 10,000 × g for 10 min at 4°C until no precipitate is observed, thereby obtaining the fourth centrifuge.
[0055] 5) The supernatant of the fourth centrifugation liquid was removed and filtered through a 0.45 μm and a 0.22 μm filter membrane using a 1 mL syringe (without the needle) to obtain a filtrate;
[0056] 6) Transfer the filtrate to a fifth centrifuge tube (ultracentrifuge tube) and ultracentrifuge at 100,000 × g for 90 min at 4°C to obtain the fifth centrifuge solution;
[0057] 7) After removing the supernatant of the fifth centrifugation solution and resuspending the solution in PBS, ultracentrifuge at 100,000 × g for 90 minutes at 4°C to obtain the sixth centrifugation solution;
[0058] 8) The supernatant of the sixth centrifugation solution was removed and the mixture was resuspended in PBS to obtain an ASNPs nanoparticle solution.
[0059] 3. Purification of ASNPs
[0060] 1) Add pre-cooled 1× PBS to the volume of the ASNPs nanoparticle solution to 6 mL to obtain a suspension;
[0061] 2) Prepare sucrose solutions of 8%, 15%, 30%, 45%, and 60% by mass as follows:
[0062] 8% sucrose solution: 0.8 g sucrose + 10 mL PBS;
[0063] 15% sucrose solution: 1.5 g sucrose + 10 mL PBS;
[0064] 30% sucrose solution: 3 g sucrose + 10 mL PBS;
[0065] 45% sucrose solution: 4.5 g sucrose + 10 mL PBS;
[0066] 60% sucrose solution: 6 g sucrose + 10 mL PBS;
[0067] 3) Add 6 mL of 8%, 15%, 30%, 45%, and 60% sucrose solutions from the bottom (60% is at the bottom). Let stand for 30 minutes. The sucrose solutions of different concentrations will form layers due to gravity with clear dividing lines. Carefully add 6 mL of the suspension dropwise to the upper layer of the 8% sucrose solution. Ultracentrifuge at 150,000 g for 120 minutes at 4°C.
[0068] 4) Carefully aspirate the nanoparticles from the 30% sucrose solution and 45% sucrose solution layers, then add an appropriate amount of PBS. Ultracentrifuge at 150,000g for 120 minutes at 4°C. Resuspend in 1 mL of PBS to obtain the purified exosome suspension, which is the ASNP nanoparticles. Store at 4°C or -80°C.
[0069] At the same time, the nanoparticles were extracted using conventional methods and freeze-dried (reference: Trenkenschuh E, Friess W. Freeze-drying of nanoparticles: How to overcome colloidal instability by formulation and process optimization. Eur J Pharm Biopharm. 2021Aug; 165: 345-360. doi: 10.1016 / j.ejpb.2021.05.024.).
[0070] 4. Morphological investigation of ASNPs nanoparticles
[0071] The nanoparticles obtained by conventional methods and the ASNPs nanoparticles prepared in this example were photographed by electron microscopy for comparison;
[0072] ASNPs nanoparticles were added with 2% silicotungstic acid for staining for 3 minutes, washed with pure water for 3 minutes, and naturally air-dried at room temperature. They were then placed under a transmission electron microscope at a voltage of 100 kV for imaging.
[0073] 5. ASNPs nanoparticle size and potential analysis
[0074] The obtained ASNPs nanoparticles were diluted 250,000 times with PBS solution, and 1 mL of the diluted solution was dropped into a nanoparticle tracking analyzer (Particle Metrix, model Zetaview-PMX120-Z) for detection.
[0075] 6. Effects of ASNPs on Vascular Endothelial and Fibroblast Aging Models
[0076] 6.1 Cell culture
[0077] Human umbilical vein endothelial cells (HUVECs) were cultured in ECM medium containing 10% FBS and 1% (100 IU / mL) streptomycin-penicillin at 37°C in an incubator with 5% CO2. NIH / 3T3 fibroblasts were cultured in DMEM medium containing 10% newborn calf serum and 1% (100 IU / mL) streptomycin-penicillin at 37°C in an incubator with 5% CO2.
[0078] 6.2 Cell seeding
[0079] The culture medium was discarded, the cells were washed once with PBS, 1 mL of trypsin was added, digested, 1 mL of culture medium was added to terminate the digestion, the suspension was pipetted evenly, the cell suspension was aspirated into a 5 mL centrifuge tube, and centrifuged at 1000 rpm for 3 min. After completion, the supernatant was discarded, 1 mL of culture medium was added to resuspend, the suspension was pipetted evenly, 20 μL of cell suspension was aspirated into a cell counting plate, and the cells were counted. Calculate 30 wells, 1.5 × 10 5 cells, a total of 6000 μL cell suspension, seeded the plate, and cultured overnight.
[0080] 6.3 Administration
[0081] HUVEC cells were treated with 100 nM doxorubicin (DOX) for 2 h as a senescence-induced cell model, and NIH / 3T3 cells were treated with 30 nM doxorubicin (DOX) for 2 h as a senescence-induced cell model. Both groups of models were treated with ASNPs particles (an appropriate amount of ASNPs particles (the amount added in this example was 1.25 × 10 8 At the same time, senescent model cells and normal cells were treated with equal amounts of PBS solution as the model group (DOX) and the control group (control).
[0082] 6.4 β-Galactosidase staining
[0083] Follow the instructions for the β-galactosidase staining kit (Biyuntian C0602). Aspirate the cell culture medium, wash once with PBS or HBSS, add 1 mL of β-galactosidase staining fixative, and fix at room temperature for 15 minutes. Aspirate the cell fixative and wash the cells three times with PBS. Aspirate the PBS and add 100 μL of staining solution to each well. Incubate at 37°C overnight. Discard the staining solution, add 100 μL of PBS to each well, and store at 4°C.
[0084] 6.5 Microscopic examination and photography
[0085] The staining results of the above steps were photographed using the THUNDER wide-field high-resolution imaging system.
[0086] 6.6 Quantitative Calculation
[0087] Image J software was used to quantify the average area of β-galactosidase-positive regions and the area occupied by cells in the visual field in each group, and the ratio of the two was used to calculate the percentage of β-galactosidase-positive staining.
[0088] 7. RT-qPCR Experiment
[0089] Male 4-month-old C57 / BL6J mice were randomly divided into four groups, with 6 mice in each group: control group, ASNPs particle treatment group (1×10 8 , 3×10 8 , 9×10 8 / mL). The mice were adaptively raised for 1 week, and the back skin was removed and prepared. The corresponding concentration of ASNPs particle hydrogel was applied once a day for 4 consecutive weeks. After the last application, the mice were killed by separating the cervical vertebrae. A back skin sample was taken from each mouse. 20-30 mg of mouse skin samples were weighed from each group, ground and homogenized using Trizol reagent, and total RNA was extracted. The RNA concentration was determined by NanoDrop 2000 spectrophotometer, and reverse transcribed into cDNA according to the reverse transcription kit HiScriptIIQ RT SuperMixfor qPCR instructions. Quantitative real-time polymerase chain reaction was performed using the 2×ChamQ SYBR qPCR Master Mix kit, with 2 -ΔΔCt The relative mRNA expression levels were calculated by the PCR method. Primers were synthesized by Shanghai Sangon Biotechnology Co., Ltd. The primer sequences are shown in Table 1.
[0090] Table 1 Primer names and sequences
[0091]
[0092]
[0093] 8. Statistical analysis
[0094] GraphPad Prism 9 software was used for statistical analysis of experimental results. Data were expressed as mean ± standard deviation (mean ± SD). One-way ANOVA was used for comparisons between multiple groups, and unpaired t-test was used for comparisons between two groups. P ≤ 0.05 indicated statistical significance. # indicates comparison with the control group, P ≤ 0.05; * indicates comparison with the model group, P ≤ 0.05; ns indicates comparison with the control group, P > 0.05.
[0095] 9. Results
[0096] 9.1 Morphological Characterization of ASNPs
[0097] Depend on Figure 1It can be seen that the conventional method cannot obtain complete nanoparticles, while the nanoparticles obtained by the method provided by the present invention are uniformly spherical; the average diameter is about 175.7±68.7nm ( Figure 2 ).
[0098] 9.2ASNPs Nanoparticle Size and Potential Analysis
[0099] Depend on Figure 3 It can be seen that the particle size of ASNPs derived from Astragalus is 169.1±64.5nm, and the particle size of Salvia miltiorrhiza derived from particles is 164.8±60.8nm; Figure 4 It can be seen that among the ASNPs nanoparticles, the Zeta potential of the particles derived from Astragalus is -39.154mV, and the Zeta potential of the particles derived from Salvia miltiorrhiza is -38.388mV.
[0100] 9.3 Effects of ASNPs on Vascular Endothelial and Fibroblast Aging Models
[0101] Figure 5 The staining results of ASNPs on human vascular endothelial cell aging model and the positive percentage of β-galactosidase staining are shown in Figure 2. Figure 6 The staining results of ASNPs on the fibroblast senescence model and the positive percentage of β-galactosidase staining show that doxorubicin successfully induced the senescent cell model, and the senescence of human vascular endothelial cells and fibroblasts can be significantly alleviated after treatment with ASNPs.
[0102] 9.4 RT-qPCR results
[0103] Depend on Figure 7 It can be seen that ASNPs promote the transcription of type I collagen encoding gene Col1a1 in the skin of 4-month-old mice.
[0104] Example 2 Effect of ASNPs Nanoparticles on Nematode Survival Rate
[0105] 1. Maintenance and synchronization of C. elegans strains
[0106] Bristol strain N2 was used as the wild-type strain. Nematodes were maintained and cultured under standard conditions (20°C) on nematode growth medium (NGM) agar coated with a lawn of live Escherichia coli OP50 as a nutrient source. The day before the experiment, pregnant hermaphroditic adults were bleached by sodium hypochlorite (NaOH / HClO) treatment, eggs were purified by centrifugation, and incubated overnight in M9 buffer to obtain an age-synchronized population of first-stage (L1) nematodes. Three days after L1 synchronization, synchronized fourth-stage (L4) nematodes were obtained.
[0107] 2. ASNPs nanoparticle treatment
[0108] To evaluate the potential protective effect of ASNPs against oxidative stress induced by endogenous superoxide radical generator PQ, synchronized L4 nematodes were divided into control group (PBS) and ASNPs nanoparticle treatment group (1×10 8 / mL). The nematodes were transferred to NGM plates containing 50mM 5-fluoro-2′-deoxyuridine (FuDR) to block offspring production. Synchronized L4 nematodes were cultured with ASNPs for 48 hours under a single diet (OP50 suspension). Each group had approximately 30 nematodes per plate, for a total of 100-130 individuals per group. Unresponsive nematodes were determined to be dead by repeated gentle mechanical stimulation (touch) and removed from the culture dish. Survival was assessed at the endpoint.
[0109] 3. Results
[0110] The survival rates of nematodes in the two groups were shown in Figure 8 The nematodes treated with ASNPs were able to resist the damage caused by oxygen free radicals, and their survival period was extended from 19 days to 21 days, a 10.5% extension.
[0111] Based on the above experimental results, the present invention provides a method for extracting and preparing ASNPs nanoparticles. This method can successfully extract Astragalus and Salvia miltiorrhiza nanoparticles (ASNPs). Cell and animal experiments have confirmed that the ASNPs prepared by the present invention can significantly alleviate cellular aging, resist oxidative stress induced by the endogenous superoxide free radical generator PQ, and improve the survival rate of nematodes. Therefore, the present invention provides a new approach and technical foundation for alleviating aging and resisting oxidative stress. The ASNPs prepared by the present invention have broad application prospects.
[0112] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.
Claims
1. A method for preparing nanoparticles, characterized in that: The following steps are involved: After adding PBS, astragalus and salvia miltiorrhiza were squeezed and filtered to obtain astragalus and salvia miltiorrhiza stock solution; The Astragalus and Salvia miltiorrhiza stock solution was centrifuged six times to obtain a nanoparticle solution; The nanoparticle solution is purified to obtain nanoparticles.
2. The preparation method according to claim 1, wherein The six centrifugal operations are as follows: Centrifuging the Astragalus and Salvia miltiorrhiza stock solution at 1000×g for 15 minutes at 4° C. to obtain a first centrifuge solution; Centrifuge the supernatant of the first centrifuge again at 3000×g for 30 min at 4°C to obtain a second centrifuge; The supernatant of the second centrifuge was centrifuged again at 10,000 × g for 30 min at 4°C to obtain a third centrifuge; The supernatant of the third centrifuge was centrifuged again at 10,000 × g for 10 min at 4° C. to obtain a fourth centrifuge; Filtering the supernatant of the fourth centrifuge through a 0.45 μm filter membrane and a 0.22 μm filter membrane to obtain a filtrate; The filtrate was ultracentrifuged at 100,000×g for 90 minutes at 4° C. to obtain a fifth centrifuge; The precipitate of the fifth centrifuge was resuspended, and ultracentrifuged at 100,000 × g for 90 min at 4° C. to obtain a sixth centrifuge; The precipitate of the sixth centrifugation liquid is resuspended to obtain a nanoparticle solution.
3. The preparation method according to claim 1, wherein The mass ratio of the astragalus root to the salvia miltiorrhiza root is 1.5:1-2.5:
1.
4. The preparation method according to claim 1, wherein The juice squeezing operation was as follows: squeezing for 1 minute, stopping for 2 minutes, and squeezing for 7 times in total.
5. The preparation method according to claim 1, wherein The purification operation is as follows: adding the nanoparticle solution to a sucrose density gradient solution and centrifuging; the sucrose density gradient solution is 8%, 15%, 30%, 45% and 60% sucrose solution; The nanoparticles in the 30% sucrose solution and 45% sucrose solution layers were aspirated, and resuspended by centrifugation to obtain ASNPs nanoparticles.
6. The preparation method according to claim 5, wherein The centrifugation conditions were all at 4° C., 150,000 g ultracentrifugation for 120 min.
7. Nanoparticles obtained by the preparation method according to any one of claims 1 to 6.
8. Use of the nanoparticles according to claim 7 in the preparation of anti-aging products.
9. An anti-aging product, characterized in that: Comprising the nanoparticles according to claim 7.
10. The product according to claim 9, characterized in that Pharmaceutically acceptable excipients are also included.