Application of naked flower extractive in preparation of anti-aging product

Abstract

This invention belongs to the field of biomedical technology, specifically relating to the application of *Callicarpa nudiflora* extract in the preparation of anti-aging products. Experiments revealed that *Callicarpa nudiflora* extract can significantly improve the survival rate and lifespan of nematodes, and also enhance their health, such as body movement ability, demonstrating a significant anti-aging effect. Furthermore, experiments using doxorubicin-induced premature aging mice as an animal model showed that *Callicarpa nudiflora* extract can significantly reduce the expression of aging-related pro-inflammatory cytokines, inflammatory chemokines, and cell cycle inhibitory factors, thereby achieving an anti-aging effect. This anti-aging effect is similar to, or even better than, the positive control metformin, making it highly suitable for the preparation of anti-aging products.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology. More specifically, it relates to the application of extracts from *Callicarpa nudiflora* in the preparation of anti-aging products. Background Technology

[0002] Aging refers to the gradual decline of physiological functions after the body reaches maturity, eventually leading to death. This decline in bodily functions can induce various age-related diseases, such as diabetes, cardiovascular disease, osteoporosis, atherosclerosis, and Alzheimer's disease. Therefore, delaying aging and maintaining health and longevity makes the development and research of anti-aging drugs of great significance and value. Currently, many drugs used clinically for anti-aging, such as metformin, acarbose, and rapamycin, are mostly chemical drugs. These older drugs lack structural novelty, and long-term use poses certain safety risks.

[0003] Many traditional Chinese medicines (TCMs) also possess anti-aging effects. For example, Chinese patent application CN114073735A discloses an anti-aging TCM composition comprising wolfberry, safflower, mulberry, hawthorn, and pearl powder, prepared through ethanol extraction. This composition increases the thymus and spleen indices in a D-galactose-induced mouse aging model, enhances serum SOD and GSHPx activity, reduces MDA levels in brain tissue, and inhibits lipid peroxidation in serum and tissues, thus achieving an anti-aging effect. It is evident that TCMs or TCM compositions can achieve anti-aging effects comparable to chemical drugs, and many TCM herbs have not been fully explored and utilized in the health industry, necessitating the development of more anti-aging active ingredients derived from TCMs. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of existing chemical drugs with certain toxic side effects, and to provide a safe and effective application of Callicarpa nudiflora extract in the preparation of anti-aging products.

[0005] Another object of the present invention is to provide the use of Callicarpa nudiflora extract in the preparation of drugs that improve the survival rate and / or prolong the lifespan of nematodes.

[0006] The above-mentioned objective of this invention is achieved through the following technical solution:

[0007] Application of Callicarpa nudiflora extract in the preparation of anti-aging products.

[0008] Furthermore, the extract of *Callicarpa nudiflora* is an ethanol extract of *Callicarpa nudiflora*.

[0009] Furthermore, the ethanol extract of Callicarpa nudiflora is obtained by extracting Callicarpa nudiflora with a 70-80% aqueous ethanol solution.

[0010] Furthermore, the preparation method of the Naked Flower Purple Callicarpa extract specifically includes the following steps:

[0011] Extract the naked flower of Callicarpa nudiflora with 70-80% ethanol aqueous solution 1-3 times, each time for 8-12 hours, combine the filtrates, concentrate and dry to obtain the product.

[0012] Preferably, the mass-to-volume ratio of the naked-flowered artichoke to the 70-80% ethanol aqueous solution is 1:(5-25) kg / L.

[0013] Preferably, the concentration is vacuum concentration and the drying is freeze drying.

[0014] Preferably, the extraction is carried out at room temperature, specifically at a temperature of 15–35°C.

[0015] Furthermore, the extraction of Callicarpa nudiflora with 70-80% ethanol aqueous solution also includes an ethyl acetate extraction step.

[0016] Furthermore, the preparation method of the Naked Flower Purple Callicarpa extract specifically includes the following steps:

[0017] Extract the naked flower of Callicarpa nudiflora with 70-80% ethanol aqueous solution 1-3 times, each time for 8-12 hours, combine the filtrates, concentrate and dry; redissolve in water, and then extract with ethyl acetate 2-4 times, each time for 30-120 minutes, collect the ethyl acetate fraction, concentrate and dry to obtain the final product.

[0018] Preferably, the resolution is performed using ultrasound-assisted dissolution.

[0019] Furthermore, the anti-aging product is characterized as a pharmaceutical, food, health product, cosmetic, feed, or additive.

[0020] Furthermore, the extract of Callicarpa nudiflora reduces the expression of aging markers TNF-α, IL-1β, CXCL-1 and p21.

[0021] Furthermore, based on the results of nematode experiments, this invention also claims protection for the use of Callicarpa nudiflora extract in the preparation of drugs that improve nematode survival rate and / or prolong nematode lifespan.

[0022] Furthermore, the extract of *Callicarpa nudiflora* enhances the body-shaking ability of nematodes.

[0023] The present invention has the following beneficial effects:

[0024] This invention, through experiments, has found that extracts of *Callicarpa nudiflora* can significantly improve the survival rate and lifespan of nematodes, and also enhance their health, such as body movement ability, demonstrating significant anti-aging effects. Furthermore, experiments using doxorubicin-induced premature aging mice as an animal model showed that *Callicarpa nudiflora* extract can significantly reduce the expression of aging-related pro-inflammatory cytokines, inflammatory chemokines, and cell cycle inhibitory factors, thereby achieving an anti-aging effect. The anti-aging effect is similar to, or even superior to, metformin as a positive control. Moreover, continuous administration for more than 20 days showed no toxic reactions, indicating high safety, making it highly suitable for the preparation of anti-aging products. Attached Figure Description

[0025] Figure 1 This is a high-performance liquid chromatography chromatogram of the 75% ethanol extract of Callicarpa nudiflora leaves in Example 3 of the present invention.

[0026] Figure 2 This is a high-performance liquid chromatography chromatogram of the petroleum ether extract of Callicarpa nudiflora leaves in Example 4 of the present invention.

[0027] Figure 3 This is a high-performance liquid chromatography chromatogram of the ethyl acetate extract of Callicarpa nudiflora leaves in Example 4 of the present invention.

[0028] Figure 4 This is a high-performance liquid chromatography chromatogram of the n-butanol extract of Callicarpa nudiflora leaves in Example 4 of the present invention.

[0029] Figure 5 This is a high-performance liquid chromatography chromatogram of the aqueous extract of Callicarpa nudiflora leaves in Example 4 of the present invention.

[0030] Figure 6 The life curve of nematodes under the action of 75% ethanol extract of Callicarpa nudiflora leaves in Example 5 of the present invention (****P<0.0001).

[0031] Figure 7 This is a statistical graph showing the body movement data of nematodes under the action of 75% ethanol extract of Callicarpa nudiflora leaves in Example 6 of the present invention (****P<0.0001).

[0032] Figure 8 The life curve of nematodes under the action of 75% ethanol extract of Callicarpa nudiflora leaves in Example 7 of the present invention (*P<0.05).

[0033] Figure 9 This is a life curve of nematodes under the action of petroleum ether extract of Callicarpa nudiflora leaves in Example 7 of the present invention.

[0034] Figure 10The life curve of nematodes under the action of ethyl acetate extract of Callicarpa nudiflora leaves in Example 7 of the present invention is shown (*P<0.05, ****P<0.0001).

[0035] Figure 11 This is a life curve of nematodes under the action of n-butanol extract of Callicarpa nudiflora leaves in Example 7 of the present invention.

[0036] Figure 12 This is a life curve of nematodes under the action of the aqueous extract of Callicarpa nudiflora leaves in Example 7 of the present invention.

[0037] Figure 13 This is a statistical chart showing the effects of 75% ethanol extract and ethyl acetate extract of *Callicarpa nudiflora* leaves on SA-β-Gal in the kidneys of an adriamycin-induced premature aging mouse model, as described in Example 8 of this invention.

[0038] Figure 14 The following is a statistical graph showing the effects of the 75% ethanol extract and the ethyl acetate extract of *Callicarpa nudiflora* leaves on the expression of pro-inflammatory cytokines TNF-α (A), IL-1β (B), chemokine CXCL-1 (C), and the aging-related biomarker p21 in the kidneys of an adriamycin-induced premature aging mouse model (*P<0.05, **P<0.01). Detailed Implementation

[0039] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0040] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0041] Example 1: Preparation of 75% ethanol extract of Callicarpa nudiflora leaves

[0042] The leaves of the Callicarpa nudiflora plant were naturally dried. 500.0g of the leaves were weighed and soaked in 12 liters of 75% ethanol. The leaves were extracted three times at room temperature, each time overnight. The extracts were combined after filtration and concentrated under reduced pressure to obtain an extract. The extract was freeze-dried to obtain a 75% ethanol extract of Callicarpa nudiflora leaves.

[0043] Example 2: Preparation of petroleum ether, ethyl acetate, and n-butanol extracts and aqueous raffinate from Callicarpa nudiflora leaves

[0044] Weigh 10g of the 75% ethanol extract of Callicarpa nudiflora leaves from Example 1, measure 400mL of pure water, add the extract in portions, adding 1g of powder each time, and stir repeatedly under ultrasonic assistance until completely dissolved; extract with petroleum ether, ethyl acetate, and n-butanol in sequence, each extraction three times, 30min each time; after concentration and drying, obtain 0.525g of petroleum ether fraction (extraction rate 5.25%), 1.089g of ethyl acetate fraction (extraction rate 10.89%), 3.197g of n-butanol fraction (extraction rate 31.97%), and 5.052g of aqueous raffinate.

[0045] Example 3: High-performance liquid chromatography analysis of 75% ethanol extract of Callicarpa nudiflora leaves

[0046] 10 mg of the 75% ethanol extract of *Callicarpa nudiflora* leaves from Example 1 was dissolved in 1 mL of methanol (chromatographic grade) under ultrasonic assistance. The dissolved solution was filtered through a 0.22 μm filter. The ethanol extract of *Callicarpa nudiflora* leaves was analyzed using a Shimadzu LC-2030Plus high-performance liquid chromatograph and a SHIMADZU C18 column (4.6 × 250 mm HSS, 5 μm). The mobile phase was acetonitrile and water. Chromatographic conditions were as follows: 10–90% acetonitrile for 0–30 min, 90% acetonitrile for 30–40 min, flow rate 1 mL / min, column temperature 30 °C, and detection wavelength 254 nm. The results are as follows. Figure 1 As shown.

[0047] As shown in the figure, the 75% ethanol extract of Callicarpa nudiflora leaves has a high concentration of active ingredients and contains a variety of different components.

[0048] Example 4: High-performance liquid chromatography analysis of extracts from leaves of Callicarpa nudiflora using different solvents

[0049] 10 mg of samples from different solvent extracts in Example 2 were dissolved in 1 mL of methanol (chromatographic grade) under ultrasonic assistance. After dissolution, the solution was filtered through a 0.22 μm filter. Using a Shimadzu LC-2030Plus high-performance liquid chromatograph and a SHIMADZU C18 (4.6 × 250 mm HSS, 5 μm) with acetonitrile and water as the mobile phase, the petroleum ether fraction, ethyl acetate fraction, n-butanol fraction, and aqueous raffinate of the leaves of *Callicarpa nudiflora* were analyzed under the following chromatographic conditions: 10–90% acetonitrile for 0–30 min, 90% acetonitrile for 30–40 min, flow rate 1 mL / min, and detection wavelength 254 nm. The chromatograms are shown below. Figures 2-5 As shown.

[0050] As shown in the figure, the ethyl acetate fraction of Callicarpa nudiflora leaves has more active ingredient peaks, which are similar to those of the 75% ethanol extract, while the aqueous and petroleum ether fractions have fewer peaks.

[0051] Example 5: Nematode lifespan experiment using 75% ethanol extract of Callicarpa nudiflora leaves.

[0052] Wild-type *C. elegans* (N2) was used in this experiment. Synchronized nematodes were transferred at stage L4 to nematode growth medium (NGM) for the blank control group and groups treated with 75% ethanol extract of *Callicarpa nudiflora* leaves (100 μg / mL, 200 μg / mL, and 400 μg / mL), with approximately 60 nematodes in each group. The medium was changed every 3 days, and the nematodes were cultured at 20°C. The number of surviving nematodes was recorded daily, and dead nematodes were removed from the culture dishes until all nematodes died. Data were summarized, survival curves were plotted using GraphPad, and statistical significance (Log-rank test) was calculated. The survival curves are shown below. Figure 6 As shown in Table 1, the life test data are as follows.

[0053] Table 1. Results of nematode lifespan experiment using 75% ethanol extract of Callicarpa nudiflora leaves.

[0054]

[0055] It was observed that the 75% ethanol extract of *Callicarpa nudiflora* leaves at 100 μg / mL significantly prolonged the lifespan of nematodes, increasing it by 27.60% compared to the control group (P<0.0001). At 200 μg / mL, it significantly prolonged the lifespan of nematodes, increasing it by 27.76% compared to the control group (P<0.0001). At 400 μg / mL, it significantly prolonged the lifespan of nematodes, increasing it by 29.96% compared to the control group (P<0.0001).

[0056] Example 6: Nematode wobble test of 75% ethanol extract of Callicarpa nudiflora leaves

[0057] Nematodes were cultured according to the method described in Example 5 for the nematode lifespan test. On days 4 and 11 of adulthood, 20 to 30 nematodes were selected from both the control group and the 75% ethanol extract group of *Callicarpa nudiflora* leaves. The number of body movements per nematode in the buffer solution (M9) was recorded for 30 seconds. After the experiment, the data were statistically analyzed, and statistical significance was calculated using a graph pad (Two-way ANOVA, Sidak multiple comparisons test). The error bars indicate the standard error of the mean (SEM).

[0058] Experimental results are as follows Figure 7 As shown, the 75% ethanol extract of Callicarpa nudiflora leaves (100 μg / mL, 200 μg / mL, and 400 μg / mL) significantly improved the body wiggling ability of nematodes, with significant differences on both day 4 and day 11. **** indicates P < 0.0001.

[0059] Example 7: Experiment on nematode lifespan using extracts from *Callicarpa nudiflora* leaves in different solvents.

[0060] Synchronized nematodes were transferred at stage L4 to nematode growth media (NGM) containing different solvent extracts (100 μg / mL, 200 μg / mL) and 75% ethanol extract of *Callicarpa nudiflora* leaves (100 μg / mL, 200 μg / mL). The medium was changed every 3 days, and the nematodes were cultured at 20°C. The number of surviving nematodes was recorded daily, and dead nematodes were removed from the culture dishes until all nematodes died. Data were summarized, survival curves were plotted using GraphPad, and statistical significance (Log-rank test) was calculated. The survival curves are shown below. Figures 8-12 As shown in Table 2, the life test data are as follows.

[0061] Table 2. Experimental data on nematode lifespan extracted from different solvents of Callicarpa nudiflora leaf.

[0062]

[0063] It was observed that the 75% ethanol extract of Callicarpa nudiflora leaves significantly prolonged the lifespan of nematodes at 100 μg / mL, with an 8.05% increase compared to the control group (* indicates P<0.05); and at 200 μg / mL, it significantly prolonged the lifespan of nematodes by 6.83% compared to the control group (* indicates P<0.05).

[0064] The petroleum ether extract of Callicarpa nudiflora leaves at 100 μg / mL prolonged the lifespan of nematodes by 4.96% compared with the control group; at 200 μg / mL, it prolonged the lifespan of nematodes by 0.62% compared with the control group, with no significant lifespan extension effect.

[0065] The ethyl acetate extract of Callicarpa nudiflora leaves at 100 μg / mL prolonged the lifespan of nematodes by 8.21% compared with the control group (* indicates P < 0.05); at 200 μg / mL, it prolonged the lifespan of nematodes compared with the control group (**** indicates P < 0.0001).

[0066] The n-butanol extract of Callicarpa nudiflora leaves at 100 μg / mL prolonged the lifespan of nematodes by 2.45% compared with the control group; at 200 μg / mL, it prolonged the lifespan of nematodes by 5.84% compared with the control group, but the effect on prolonging lifespan was not significant.

[0067] The aqueous extract of Callicarpa nudiflora leaves at 100 μg / mL prolonged the lifespan of nematodes by 6.36% compared with the control group; at 200 μg / mL, it prolonged the lifespan of nematodes by 6.01% compared with the control group, but the effect on prolonging lifespan was not significant.

[0068] Anti-aging Experiment of 75% Ethanol Extract and Ethyl Acetate Extract of Callicarpa nudiflora Leaves in Mice

[0069] Male C57BL / 6J mice (7 weeks old) were purchased from Guangdong Vital River Laboratory Animal Technology Co., Ltd. The mouse experiment was carried out in the Experimental Animal Center of the School of Pharmacy, Hainan University, in the Industrial Incubation Park of the High-tech Zone in Haikou City, with the license number: SYXK(Qiong)2023 - 0031. According to the protocol of the reference (Tianyue Sun, Lanxin Zhang, et al. Characterization of cellular senescence in doxorubicin-induced aging mice[J]), the common chemotherapeutic drug doxorubicin was used as an inducer of premature aging and hepatotoxicity in rodents and humans. Premature aging mice induced by doxorubicin were used as animal models, and the anti-aging drug efficacy of the ethanol extract and ethyl acetate extract of Callicarpa nudiflora leaves was evaluated through biomarkers such as pro-inflammatory cytokines, inflammatory chemokines, and cell cycle inhibitors (such as TNF-α, IL-1β, CXCL-1, and p21).

[0070] In the blank group, mice were intragastrically administered normal saline; in the doxorubicin induction group, mice were intraperitoneally injected with doxorubicin at 5 mg / kg on day 0 and day 10 to induce aging, and normal saline was intragastrically administered starting from day 15; in the 75% ethanol extract treatment group of Callicarpa nudiflora leaves, doxorubicin at 5 mg / kg was also intraperitoneally injected on day 0 and day 10 to induce aging, and the 75% ethanol extract of Callicarpa nudiflora leaves was administered starting from day 15, with two concentration groups of 1 g / kg and 2 g / kg; in the ethyl acetate extract treatment group of Callicarpa nudiflora leaves, doxorubicin at 5 mg / kg was intraperitoneally injected on day 0 and day 10 to induce aging, and a dose of 1 g / kg of the ethyl acetate extract of Callicarpa nudiflora leaves was administered starting from day 15; according to the above reference, metformin was used as a positive anti-aging drug, and metformin (20 mg / kg) was set as the positive control group.

[0071] Medication was given until day 39, and on day 40, the mice in each group were dissected, and the internal organs of each part were separated and immediately cryopreserved.

[0072] 1. SA-β-Gal Staining Experiment of Mouse Kidneys

[0073] Liver and kidney samples were collected from each group of animals, embedded in embedding medium, frozen in liquid nitrogen (N2), cut into 20 μm sections, and fixed on glass slides. The slides were brought to room temperature for 5 minutes to warm up. After warming, the slides were rinsed twice in cold phosphate-buffered saline (PBS, Servicebio), 5 minutes each time. The rinsed slides were then fixed by rinsing with fixative (0.8% formaldehyde + 0.4% glutaraldehyde) for 10 minutes, followed by rinsing twice in cold phosphate-buffered saline, 5 minutes each time. Finally, the slides were placed in preheated X-gal staining solution (deionized water + citrate-disodium hydrogen phosphate buffer + 0.1M potassium ferricyanide + 0.1M potassium ferrocyanide + 5M sodium chloride + 1M magnesium chloride + X-gal solution (20 mg / mL, DMF)) at 37°C and stained for 12 hours. The staining was observed the next day, and staining was terminated by rinsing with PBS. The stained slides were then rinsed sequentially in 75% ethanol, 85% ethanol, anhydrous ethanol, and xylene, 5 minutes each time. This yielded stained mouse kidney tissue slides.

[0074] The above sections were photographed and quantitatively analyzed using a microscope, and the results are as follows: Figure 13 As shown. Quantitative analysis of mouse kidney sections using SA-β-Gal staining revealed that, compared to the doxorubicin model group, the low-concentration group (1 g / kg) of 75% ethanol extract of *Callicarpa nudiflora* leaves, the high-concentration group (2 g / kg) of 75% ethanol extract of *Callicarpa nudiflora* leaves, and the ethyl acetate extract group (1 g / kg) of *Callicarpa nudiflora* leaves all showed a trend of reversing doxorubicin-induced aging, and the effect was superior to the positive control metformin (20 mg / kg) treatment group.

[0075] 2. Mouse kidney qPCR experiment

[0076] Kidneys were harvested from mice in each group, and kidney tissue the size of a mung bean was cut and placed in 1.5 mL enzyme-free centrifuge tubes. Two 3 mm enzyme-free grinding beads and one 4 mm enzyme-free grinding bead were added. Total RNA was extracted using a total RNA extraction reagent (10606ES60, Yeasen), and qPCR was performed. First-strand cDNA was synthesized using SuperMix (11123ES10, Yeasen) and reverse transcribed into cDNA. Using... qPCR was performed using SYBR Green Master Mix (nuclease-free) (11201ES 08, Yeasen) for polymerase chain reaction (PCR) amplification to detect the mRNA expression levels of TNF-α, IL-1β, CXCL-1, and p21. The primer sequences are as follows:

[0077] TNF-αForward:5'-GCCTCTTCTCATTCCTGCTT-3'

[0078] TNF-αReverse:5'-CTCCTCCACTTGGTGGTTTG-3'

[0079] IL-1βForward:5'-TGCCACCTTTTGACAGTGATG-3'

[0080] IL-1βReverse:5'-TGATGTGCTGCTGCGAGATT-3'

[0081] CXCL-1Forward:5'-ACCGAAGTCATAGCCACACTC-3'

[0082] CXCL-1Reverse:5'-CTCCGTTACTTGGGGACACC-3'

[0083] p21 Forward:5'-CAGATCCACAGCGATATCCA-3'

[0084] p21 Forward:5'-ACGGGACCGAAGAGACAAC-3'

[0085] Gadph Forward:5'-GTGGCAAAGTGGAGATTGTTG-3'

[0086] Gadph Reverse:5'-AGTCTTCTGGGTGGCAGTGAT-3'

[0087] The results are as follows Figure 14 As shown, the doxorubicin-induced premature aging model in mice was successful. All treatment groups reversed the doxorubicin-induced upregulation of the pro-inflammatory cytokine TNF-α: the low-concentration group of 75% ethanol extract of *Callicarpa nudiflora* leaves (1 g / kg) was comparable to the positive control metformin treatment group (20 mg / kg); all treatment groups reversed the doxorubicin-induced upregulation of the pro-inflammatory cytokine IL-1β, with the low-concentration group of 75% ethanol extract of *Callicarpa nudiflora* leaves (1 g / kg) and the ethyl acetate extract of *Callicarpa nudiflora* leaves (1 g / kg) showing significantly better effects than the positive control metformin treatment group (20 mg / kg); all treatment groups reversed the doxorubicin-induced upregulation of the chemokine CXCL-1; and all treatment groups reversed the doxorubicin-induced upregulation of the aging-related marker P21, with significant differences between the low-concentration group of 75% ethanol extract of *Callicarpa nudiflora* leaves (1 g / kg) and the ethyl acetate extract group of *Callicarpa nudiflora* leaves (1 g / kg). * indicates P < 0.05, ** indicates P < 0.01.

[0088] In summary, the results of the nematode lifespan experiment show that the 75% ethanol extract and ethyl acetate extract of *Callicarpa nudiflora* leaves obtained in this invention can significantly prolong the lifespan of nematodes. The health parameter evaluation experiment confirmed that the 75% ethanol extract of *Callicarpa nudiflora* leaves can improve the body-shaking ability of nematodes. The SA-β-Gal aging staining experiment on mouse kidney sections showed that both the 75% ethanol extract and ethyl acetate extract of *Callicarpa nudiflora* leaves tended to improve doxorubicin-induced kidney aging in mice. The qPCR experiment on mouse kidney tissue showed that each treatment group exhibited a certain trend of reversing doxorubicin-induced premature aging in mice, with significant differences. Therefore, *Callicarpa nudiflora* has significant anti-aging effects and high utilization value in the field of anti-aging technology.

[0089] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. Use of an ethyl acetate extract of S. nudiflora in the preparation of an anti-aging pharmaceutical product, characterized in that, The preparation method of the ethyl acetate extract of the naked flower, *Callicarpa nudiflora*, specifically includes the following steps: Extract the naked flower of Callicarpa nudiflora with 75% ethanol aqueous solution 1-3 times, each time for 8-12 hours, combine the filtrates, concentrate and dry; redissolve in water, and extract with ethyl acetate 2-4 times, each time for 30-120 minutes, collect the ethyl acetate fraction, concentrate and dry to obtain the product; The ethyl acetate extract of *Callicarpa nudiflora* reduced the expression of aging markers TNF-α, IL-1β, CXCL-1, p21, and SA-β-Gal.

2. Use of the ethyl acetate extract of V. nudiflora in the preparation of anti-aging drugs, characterized in that, The preparation method of the ethyl acetate extract of the naked flower, *Callicarpa nudiflora*, specifically includes the following steps: Extract the naked flower of Callicarpa nudiflora with 75% ethanol aqueous solution 1-3 times, each time for 8-12 hours, combine the filtrates, concentrate and dry; redissolve in water, and extract with ethyl acetate 2-4 times, each time for 30-120 minutes, collect the ethyl acetate fraction, concentrate and dry to obtain the product; The ethyl acetate extract of the naked flower purple artichoke improves the survival rate of nematodes and / or prolongs their lifespan. The ethyl acetate extract of the naked flower, *Callicarpa nudiflora*, enhances the body-shaking ability of nematodes.

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