Neural cell senescence resisting medicine containing p-hydroxybenzaldehyde and application of nerve cell senescence resisting medicine

Abstract

The invention discloses a nerve cell senescence resisting medicine containing p-hydroxybenzaldehyde and application thereof.The p-hydroxybenzaldehyde is adopted as the only active substance, and the p-hydroxybenzaldehyde can reduce accumulation of DNA damage in cells, reduce activity of beta-galactosidase in the cells and slow down reduction of cell metabolic capacity, so that the nerve cell senescence resisting medicine is obtained. Therefore, the senescence of nerve cells is effectively slowed down, the cognitive function is improved, and a potential treatment effect is achieved in neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology. Specifically, it relates to an anti-neural cell aging drug containing p-hydroxybenzaldehyde and its application. Background Art

[0002] As humans age, the cognitive function of the brain gradually declines due to brain aging, which is closely related to the aging of brain nerve cells. The aging of brain nerve cells is an extremely complex terminal cell state, manifested by various pathophysiological changes: continuous accumulation of DNA damage leads to impaired integrity of genetic material, the activity of the cell senescence marker β-galactosidase increases significantly, and at the same time, the energy metabolism and material synthesis capabilities of the cells are significantly reduced. These cell-level aging characteristics will further cause functional disorders such as decreased synaptic plasticity of nerve cells and disordered neurotransmitter secretion, ultimately leading to abnormal organizational structures in key brain regions such as the hippocampus and prefrontal cortex and reduced neural network connection efficiency.

[0003] p-Hydroxybenzaldehyde (p-HBA), as a non-flavonoid phenolic compound, is widely present in medicinal plants such as Gastrodia elata, Galeola septentrionalis, and Vanilla planifolia. Previous studies have confirmed its significant antifungal biological activity. However, in the important medical field of anti-brain aging, there are currently no relevant research reports on p-hydroxybenzaldehyde delaying the process of brain aging through pathways such as clearing senescent cells, repairing DNA damage, or improving neuronal metabolism, which provides a potential research direction for the development of new neuroprotective drugs.

[0004] In view of this, the present invention is specifically proposed. Summary of the Invention

[0005] The purpose of the present invention is to provide an anti-neural cell aging drug containing p-hydroxybenzaldehyde and its application, wherein p-hydroxybenzaldehyde can delay the aging process of nerve cells through pathways such as clearing senescent cells, repairing DNA damage, or improving neuronal metabolism.

[0006] The present invention is implemented as follows:

[0007] The present invention provides an anti-neural cell aging drug containing p-hydroxybenzaldehyde. p-Hydroxybenzaldehyde is used as the only active substance, and its structural formula is shown in formula (I):

[0008]

[0009] In some preferred embodiments, the neural cell aging is induced by hydroxyurea.

[0010] In some preferred embodiments, the nerve cells are human neuroblastoma cells SH-SY5Y.

[0011] In some preferred embodiments, the drug is a preparation prepared from p-hydroxybenzaldehyde as an active ingredient, added with a pharmaceutically acceptable carrier and / or excipient.

[0012] In some preferred embodiments, the preparation is an oral preparation or an injection preparation.

[0013] The present invention also provides an application of an anti-nerve cell aging drug containing p-hydroxybenzaldehyde in preventing or treating nerve cell aging-related diseases.

[0014] In some preferred embodiments, the application of the drug in reducing the phosphorylation level of H2AX protein.

[0015] In some preferred embodiments, the application of the drug in reducing the activity of β-galactosidase.

[0016] In some preferred embodiments, the application of the drug in increasing the protein levels of ATG-3 and ATG-4.

[0017] In some preferred embodiments, the application of the drug in increasing the protein level of LC3-II and reducing the protein level of p62.

[0018] The present invention has the following beneficial effects:

[0019] P-hydroxybenzaldehyde effectively slows down the aging of nerve cells, improves cognitive function, and plays a potential therapeutic role in neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease by reducing the accumulation of DNA damage in cells, reducing the activity of β-galactosidase in cells, and slowing down the decrease in cell metabolic capacity. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings required to be used in the embodiments. It should be understood that the following drawings only show some embodiments of the present invention, and therefore should not be regarded as limiting the scope. For those of ordinary skill in the art, without creative efforts, other related drawings can also be obtained based on these drawings.

[0021] Figure 1 For the toxicity determination of p-hydroxybenzaldehyde;

[0022] Figure 2Effect of p-hydroxybenzaldehyde on the β-galactosidase activity of hydroxyurea-induced SH-SY5Y cells; A: Microscopic imaging, blue-stained cells are positive cells; B: Positive rate of β-galactosidase staining in each group;

[0023] Figure 3 Effect of p-hydroxybenzaldehyde on DNA damage of hydroxyurea-induced SH-SY5Y cells; A: Staining results of immunofluorescence detection of phosphorylated H2AX positive cells; B: Proportion of phosphorylated H2AX positive cells detected by immunofluorescence;

[0024] Figure 4 Effect of p-hydroxybenzaldehyde on the autophagy function of hydroxyurea-induced SH-SY5Y cells; A: Western blot of LC3-I and LC3-II proteins; B: Quantitative results of LC3-I and LC3-II proteins; C: Western blot of p62 protein; D: Quantitative results of p62 protein; E: Western blot of ATG-3 protein; F: Quantitative results of ATG-3 protein;

[0025] Figure 5 Effect of p-hydroxybenzaldehyde on the autophagosome formation of hydroxyurea-induced SH-SY5Y cells; A: Immunofluorescence image of autophagosomes; B: Quantitative results of autophagosomes;

[0026] Figure 6 Effect of p-hydroxybenzaldehyde on cellular senescence in the hippocampus of a D-gal-induced senile mouse model. Specific implementation mode

[0027] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Those not specified in the embodiments are carried out according to conventional conditions or conditions recommended by the manufacturer. Reagents or instruments not specified by the manufacturer are all conventional products that can be obtained through commercial purchase.

[0028] The following specifically describes an anti-neural cell senescence drug containing p-hydroxybenzaldehyde and its application proposed in this application.

[0029] This application provides an anti-neural cell senescence drug containing p-hydroxybenzaldehyde, and p-hydroxybenzaldehyde is used as the only active substance, and its structural formula is shown in formula (I):

[0030]

[0031] p-Hydroxybenzaldehyde (p-HBA) exhibits various beneficial effects in anti-neuronal cell aging, mainly acting through its antioxidant, anti-inflammatory, and neuroprotective mechanisms. As a natural phenolic compound, p-Hydroxybenzaldehyde can effectively scavenge free radicals, reduce the damage of oxidative stress to neuronal cells, and simultaneously activate the Nrf2 / ARE pathway, upregulating the expression of endogenous antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GPx), thereby enhancing the antioxidant defense ability of cells.

[0032] Furthermore, p-Hydroxybenzaldehyde can effectively alleviate hydroxyurea-induced cell aging and reduce the accumulation of toxic substances such as β-amyloid (Aβ), thereby reducing its damage to neurons and protecting the functional integrity of nerve cells. Secondly, p-Hydroxybenzaldehyde can repair DNA damage caused by hydroxyurea, specifically manifested as a significant reduction in the phosphorylation level of H2AX, indicating its important role in maintaining genomic stability and contributing to delaying the cell aging process. In addition, p-Hydroxybenzaldehyde exerts an anti-aging effect by promoting autophagy function, and its mechanisms include upregulating the protein level of ATG-3, promoting autophagosome formation, and enhancing autophagic flux, thereby reversing the autophagy dysfunction caused by hydroxyurea. These effects together indicate that p-Hydroxybenzaldehyde can not only multi-targetedly intervene in the key pathways of neuronal cell aging but also delay the aging process by enhancing the cell self-repair ability, providing a potential new strategy for the development of drugs against neurodegenerative diseases.

[0033] In some preferred embodiments, human neuroblastoma cell line SH-SY5Y was used as the cell model, and hydroxyurea (HU) was used as the drug to induce cell aging.

[0034] Among them, human neuroblastoma cell line SH-SY5Y is a neuronal-like tumor cell line with the characteristics of nerve cells. The aging model established using this cell is closer to the aging process of nerve cells in the human body and has more reference value in physiological research. In addition, as a drug to induce cell aging, hydroxyurea can inhibit the activity of ribonucleotide reductase, thereby hindering DNA synthesis and disrupting cell growth and metabolism. In previous studies, hydroxyurea has been used as an inducer of cell aging in various cells, such as neural stem cells and fibroblasts. Using hydroxyurea as an aging inducer to construct a cell aging model helps to discover how cells respond to DNA damage and observe the process of DNA repair, better verifying the role of p-Hydroxybenzaldehyde in combating brain aging.

[0035] In some preferred embodiments, the drug is a preparation prepared with p-hydroxybenzaldehyde as an active ingredient, plus a pharmaceutically acceptable carrier and / or excipient, and the preparation is an oral preparation or an injection preparation.

[0036] The present application also provides an application of an anti-neural cell senescence drug containing p-hydroxybenzaldehyde in preventing or treating neural cell senescence-related diseases.

[0037] In some preferred embodiments, the drug can be applied to reduce the phosphorylation level of H2AX protein, reduce the activity of β-galactosidase, increase the protein levels of ATG-3 and ATG-4, and increase the protein level of LC3-II and reduce the protein level of p62.

[0038] The features and properties of the present invention will be further described in detail below in conjunction with examples.

[0039] In this example, the human neuroblastoma cell line SH-SY5Y used was derived from the American Type Culture Collection (ATCC number: CRL-2266).

[0040] Example 1: Toxicity determination of p-hydroxybenzaldehyde.

[0041] First, SH-SY5Y cells were cultured in DMEM / F12 medium containing 10% fetal bovine serum (FBS) in a 5% carbon dioxide incubator at 37°C. Then, the SH-SY5Y cells were digested and counted, and inoculated into a 96-well plate at a density of 5×10 4 cells / well, a total of 10 groups, including: blank group, p-HBA 0.01 μM, p-HBA 0.03 μM, p-HBA 0.1 μM, p-HBA 0.3 μM, p-HBA 1 μM, p-HBA 3 μM, p-HBA 10 μM, p-HBA 30 μM, and p-HBA 100 μM. And the medium was changed from DMEM / F12 containing 10% FBS to DMEM / F12 containing 0.5% FBS, and cultured in an incubator for 24 hours. Then, p-hydroxybenzaldehyde solutions with corresponding concentrations were prepared according to the groups and dissolved with DMEM / F12 containing 0.5% FBS. The medium in the 96-well plate was changed to the corresponding p-hydroxybenzaldehyde solution and cultured in an incubator for 48 hours. After 48 hours, the cells were incubated in 10% CCK-8 solution at 37°C for 0.5 hours. The absorbance at 450 nm was measured with a SpectraMax M5 microplate reader. All values were normalized to the control group.

[0042] The results are as Figure 1 shown. The toxicity of p-hydroxybenzaldehyde was determined using the CCK-8 solution, and it was found that the p-hydroxybenzaldehyde solution had no obvious toxicity to SH-SY5Y cells at a concentration of 0.01 - 100 μM.

[0043] Example 2: Effect of p-hydroxybenzaldehyde on β-galactosidase in SH-SY5Y cells induced by hydroxyurea.

[0044] First, SH-SY5Y cells were cultured in DMEM / F12 medium containing 10% FBS in a 5% carbon dioxide incubator at 37°C. They were seeded into six-well plates, with each well evenly covered with 1×10 5 cells, for a total of five groups, including: control group, HU group, HU + p-HBA 1 μM group, HU + p-HBA 3 μM group, and HU + p-HBA 10 μM group.

[0045] After the culture was completed, the cell culture medium was aspirated, washed once with PBS or HBSS, 1 mL of β-galactosidase staining fixative was added, and fixed at room temperature for 15 minutes. The cells were washed 3 times with PBS, 5 minutes each time. The staining solutions A, B, C, and X-Gal solution in the kit were prepared into a staining working solution according to the instructions and added to the well plates, and incubated overnight at 37°C. During observation, counting was performed under an ordinary optical microscope, and the number of stained cells was calculated.

[0046] The results were as Figure 2 shown: The test results showed that hydroxyurea (HU) treatment significantly induced the senescence of SH-SY5Y cells, manifested as a significant increase in the number of β-galactosidase (β-Gal)-positive cells, with a significant difference compared to the control group. However, when different concentrations of p-hydroxybenzaldehyde solution were added, the proportion of β-Gal-positive cells showed a dose-dependent decrease. The 1 μM p-HBA treatment group showed a certain inhibitory effect, but the effect was weak; the inhibitory effect of the 3 μM p-HBA treatment group was more obvious, and the number of β-Gal-positive cells decreased significantly; while the inhibitory effect of the 10 μM p-HBA treatment group was the most significant, almost approaching the level of the control group, indicating that high-concentration p-HBA could almost completely antagonize HU-induced cell senescence. This trend was visible in the Figure 2 quantification graph of -B, in which the proportion of β-Gal-positive cells in the HU group was the highest, and this proportion gradually decreased with the increase in the concentration of p-HBA. In summary, p-HBA could significantly alleviate HU-induced senescence of SH-SY5Y cells in the range of 1 - 10 μM, and its effect was concentration-dependent.

[0047] Example 3: Effect of p-hydroxybenzaldehyde on DNA damage in SH-SY5Y cells induced by hydroxyurea.

[0048] First, SH-SY5Y cells were cultured in DMEM / F12 medium containing 10% FBS in a 5% carbon dioxide incubator at 37°C. Then, the SH-SY5Y cells were digested and counted, and seeded into confocal dishes at a density of 2×10 5 cells / confocal dish, with a total of five groups, including: control group, HU group, HU + p-HBA 1 μM group, HU + p-HBA 3 μM group, and HU + p-HBA 10 μM group.

[0049] After 24 hours of culture, the cells were co-treated with 0.25 mM hydroxyurea and different concentrations of p-hydroxybenzaldehyde. After 48 hours, the cells were fixed with 4% PFA for 15 minutes, washed 3 times with DPBS, blocked in immunostaining blocking buffer for 1 hour, and incubated with p-H2AX antibody (1:200) overnight at 4°C. Then, the cells were incubated with Hoechst 33342 (1:1000) and secondary antibody (1:1000) in the dark for 1 hour at room temperature. After washing 3 times with DPBS, images were taken using a laser scanning microscope (Leica TCS SP8 confocal laser scanning microscope).

[0050] The results were as Figure 3 shown. p-H2AX is essential for checkpoint-mediated cell cycle arrest and DNA repair after double-strand DNA breaks, and DNA damage leads to rapid phosphorylation of H2A. Hydroxyurea treatment significantly increased the phosphorylation level of the DNA damage marker γ-H2AX in SH-SYSY cells, indicating that hydroxyurea induced obvious DNA damage. On the basis of hydroxyurea treatment, after adding different concentrations of p-hydroxybenzaldehyde solution, the phosphorylation level of γ-H2AX showed a concentration-dependent decrease. The phosphorylation level of γ-H2AX was slightly reduced in the 1 μM p-HBA treatment group, but the effect was limited; the DNA damage in the 3 μM p-HBA treatment group was significantly reduced; the effect in the 10 μM p-HBA treatment group was the most significant, and the phosphorylation level of γ-H2AX was close to the control group level. The results showed that p-hydroxybenzaldehyde could effectively inhibit hydroxyurea-induced DNA damage, and its protective effect was enhanced with increasing concentration. In addition, immunofluorescence images further confirmed the reduction of DNA damage foci in the nucleus by p-hydroxybenzaldehyde, and p-hydroxybenzaldehyde may play a protective function through antioxidant or direct DNA repair mechanisms.

[0051] Example 4: Effect of p-hydroxybenzaldehyde on the autophagy function of hydroxyurea-induced SH-SY5Y cells.

[0052] First, SH-SY5Y cells were cultured in DMEM / F12 medium containing 10% FBS in a 5% carbon dioxide incubator at 37°C. Then, the SH-SY5Y cells were digested and counted, and seeded into six-well plates at a density of 3×105 per well, with a total of five groups, including: control group, HU group, HU + p-HBA 1 μM group, HU + p-HBA 3 μM group, and HU + p-HBA 10 μM group.

[0053] After the culture was completed, the cells were washed once with ice-cold DPBS in a culture dish, and then lysed in RIPA buffer containing 1% phenylmethanesulfonyl fluoride and 1% protease / phosphatase inhibitor mixture (Thermo Fisher) at 4°C for 30 minutes. Subsequently, centrifugation was performed at 12,500 × g for 20 minutes at 4°C. The lysate in 1× sample buffer was boiled at 95°C for 5 minutes to denature the proteins, and then separated by SDS-polyacrylamide gel electrophoresis. Western blotting was performed using the corresponding specific antibody to detect the target protein, with β-actin as an internal control. According to the manufacturer's instructions, an ECL kit (GE Healthcare) was used to visualize the blot. The intensity of the bands was quantitatively analyzed using Image Lab 5.0 software.

[0054] The results are as Figure 4 shown. Autophagy is closely related to the process of cellular senescence. The autophagy process can clear the accumulated proteins and damaged organelles in cells and play a positive role in anti-aging. Chloroquine is an autophagy inhibitor that can block autophagic flux and is commonly used to detect the level of autophagic flux. Among them, Figure 4 -A Western blotting detection found that 10 μM p-hydroxybenzaldehyde could significantly restore the decrease in LC3-II protein level caused by hydroxyurea; Figure 4 -C Western blotting detection found that 10 μM p-hydroxybenzaldehyde could significantly reduce the increase in p62 protein level caused by hydroxyurea; Figure 4 -A to Figure 4 -D together indicated that p-hydroxybenzaldehyde restored the reduction in autophagic flux caused by hydroxyurea; Figure 4 -E Western blotting detection found that 10 μM p-hydroxybenzaldehyde could significantly restore the decrease in ATG-3 protein level caused by hydroxyurea; indicating that hydroxyurea caused damage to the autophagy function in cells by downregulating the ATG-3 protein level, while p-hydroxybenzaldehyde could increase the protein level of ATG-3, promote the formation of autophagosomes, and increase autophagic flux, thereby repairing the autophagy dysfunction caused by hydroxyurea.

[0055] Example 5: The effect of p-hydroxybenzaldehyde on the formation of autophagosomes in hydroxyurea-induced SH-SY5Y cells.

[0056] First, SH-SY5Y cells were cultured in DMEM / F12 medium containing 10% FBS in a 5% carbon dioxide incubator at 37°C. Then, the SH-SY5Y cells were digested and counted, and seeded into confocal dishes at a density of 2 × 105 Confocal dishes, a total of five groups, including: control group, HU group, HU + p-HBA 1 μM group, HU + p-HBA 3 μM group, HU + p-HBA 10 μM group.

[0057] Subsequently, SH-SY5Y cells were infected with mCherry-GFP-LC3 adenovirus, and autophagic flux dynamics were monitored by fluorescence labeling. In this dual-fluorescence reporting system, mCherry-GFP-LC3 emits both red (mCherry) and green (GFP) fluorescence in autophagosomes (not fused with lysosomes), presenting a yellow merged signal; when autophagosomes fuse with lysosomes, due to the acidic environment in lysosomes, GFP fluorescence is quenched, and only mCherry red fluorescence remains, thus distinguishing autophagosomes (yellow dots) from autophagolysosomes (red dots).

[0058] After the infection was completed, the cells in each group were treated with the corresponding drugs: the control group used conventional medium, the HU group was added with hydroxyurea to induce autophagy inhibition, and the other three groups were added with 1 μM, 3 μM, and 10 μM of p-HBA on the basis of HU treatment. The number and proportion of yellow (autophagosomes) and red (autophagolysosomes) spots in the cells were observed and quantified by fluorescence microscopy or confocal microscopy.

[0059] Result analysis: As Figure 5 shown, treatment with hydroxyurea led to an accumulation of yellow spots and a decrease in red spots in the cells, indicating that the transport of autophagosomes to lysosomes was blocked and autophagic flux decreased; while the addition of 10 μM p-HBA significantly increased the proportion of red spots, indicating that it promoted autophagosome maturation and fusion with lysosomes, restoring autophagic flux. This experiment verified that p-HBA alleviated hydroxyurea-induced autophagic dysfunction by enhancing autophagosome formation and subsequent degradation processes.

[0060] Example 6: Effect of p-hydroxybenzaldehyde on cellular senescence in the hippocampus of a D-galactose (D-gal)-induced senescence mouse model.

[0061] Eight-week-old female C57BL / 6 mice were randomly divided into 4 groups, including: control group, D-gal group, D-gal + p-HBA 5 mg / kg group, and D-gal + p-HBA 10 mg / kg group. The body weights were weighed before administration, and the dosage was calculated according to the body weight. Among them, the D-gal group, D-gal + p-HBA 5 mg / kg group, and D-gal + p-HBA 10 mg / kg group were subcutaneously injected with 200 mg / kg D-gal at 8 weeks of age for modeling. The drug groups were gavaged with 5 mg / kg and 10 mg / kg P-HBA starting from the 3rd week of modeling. At the 13th week of modeling, the mice were anesthetized by intraperitoneal injection of 4% chloral hydrate. The chest was quickly cut open to expose the heart. A syringe filled with normal saline was inserted into the aorta through the left ventricle. After washing the blood, 4% paraformaldehyde was used for perfusion. After the mice became stiff all over, the mouse brains were taken out, placed in 4% paraformaldehyde for fixation for 24 hours, and then transferred to 30% sucrose for dehydration in turn. After the brain completely sank to the bottom of the bottle, it was taken out and continuously coronal sectioned with a cryostat, with each section being 20 μm thick.

[0062] The brain sections were directly detected by SA-β-gal staining. The brain sections were washed twice with distilled water for 2 minutes each time, stained with SA-β-gal, incubated in the dark at 37°C for 24 hours, sealed with neutral gum, and the blue-green positive cells (senescent cells) in the hippocampal region were observed under an optical microscope.

[0063] The results were as Figure 6 shown. It was found by SA-β-gal staining that continuous administration of different concentrations of p-HBA for 13 weeks could significantly alleviate D-gal-induced senescence of hippocampal cells in mice.

[0064] In summary, p-hydroxybenzaldehyde (p-HBA) exhibits significant anti-aging effects at multiple levels. Toxicity tests indicate that p-HBA has no obvious toxicity to SH-SYSY cells within the concentration range of 0.01 - 100 μM, providing a basis for its safety. Secondly, p-HBA can alleviate hydroxyurea (HU)-induced cellular senescence in a dose-dependent manner, significantly reducing the number of β-galactosidase-positive cells and almost completely alleviating the senescence-inducing effect of HU at a concentration of 10 μM. In addition, p-HBA can effectively inhibit HU-induced DNA damage, reduce the phosphorylation level of γ-H2AX, alleviate DNA damage and promote repair. In terms of autophagy regulation, p-HBA promotes autophagosome formation and autophagic flux recovery by upregulating the protein level of ATG-3, thereby improving the autophagic dysfunction caused by hydroxyurea. Further experiments confirm that p-HBA can promote the fusion of autophagosomes and lysosomes and enhance autophagic function. Finally, animal experiments show that p-HBA can also alleviate D-gal-induced senescence of hippocampal cells in mice. In summary, p-HBA exerts anti-aging effects through multiple pathways, including inhibiting cellular senescence, reducing DNA damage, and promoting the restoration of autophagic function.

[0065] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention can have various modifications and changes. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. An anti-nerve cell aging drug containing p-hydroxybenzaldehyde, characterized in that, p-Hydroxybenzaldehyde is used as the sole active substance, and its structural formula is shown in Formula (I):

2. The anti-neural cell aging drug containing p-hydroxybenzaldehyde according to claim 1, characterized in that, The senescence of the nerve cells is induced by hydroxyurea.

3. The anti-nerve cell aging drug containing p-hydroxybenzaldehyde according to claim 1, wherein, The nerve cells are human neuroblastoma cells SH-SY5Y.

4. The anti-nerve cell aging drug containing p-hydroxybenzaldehyde according to claim 1, characterized in that The drug is a preparation prepared from p-hydroxybenzaldehyde as an active ingredient, plus a pharmaceutically acceptable carrier and / or excipient.

5. An anti-nerve cell aging drug containing p-hydroxybenzaldehyde according to claim 4, characterized in that, The preparation is an oral preparation or an injection preparation.

6. Use of an anti-nerve cell senescence drug containing p-hydroxybenzaldehyde according to any one of claims 1-5 in the prevention or treatment of nerve cell senescence-related diseases.

7. The application according to claim 6, characterized in that, Use of the drug in reducing the phosphorylation level of H2AX protein.

8. The application according to claim 6, characterized in that, Use of the drug in reducing the activity of β-galactosidase.

9. The application according to claim 6, wherein Use of the drug in increasing the protein levels of ATG-3 and ATG-4.

10. The application according to claim 6, characterized in that, Use of the drug in increasing the protein level of LC3-II and reducing the protein level of p62.

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