Application of Mianserin in delaying or treating aging

Abstract

The application belongs to the application of mianserin in delaying or treating aging, and belongs to the field of biomedicine. The technical problem solved by the application is how to treat aging. The application finds that mianserin can be used as a new drug for treating or delaying aging: at the cell level, mianserin can alleviate the aging of HGPS cells and skin fibroblasts of the elderly; at the individual level, mianserin can improve the external morphology of progeria animals, increase the body weight of progeria animals, improve the movement ability and rickets degree of progeria animals, increase the grip and food intake of progeria animals, and improve the degree of cardiac and muscle tissue fibrosis. It is shown that, as an "old drug", mianserin can be "newly used" in the treatment of aging.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to the application of mianserin in delaying or treating aging. Background Technology

[0002] Hutchinson–Gilford Progeria Syndrome (HGPS) is the most common type of progeria in children. It is a rare and fatal autosomal dominant genetic disorder. Affected children age rapidly from childhood, appearing elderly, with organ decline and decreased physiological function. Cardiovascular diseases such as myocardial infarction and heart failure, as well as systemic atherosclerotic complications, are the main causes of death in HGPS patients. The aging process in these patients is 5 to 10 times faster than normal, resembling an accelerated version of normal aging. HGPS is caused by a mutation in LMNA, which produces abnormal LaminA, or progerin. At the cellular level, progerin accumulates on the nuclear membrane, interfering with the function and normal distribution of LaminA, thereby affecting many cellular processes, such as abnormal nuclear morphology, telomere shortening, loss of peripheral heterochromatin, increased DNA damage, persistent activation of DNA damage response proteins, increased genomic instability, and premature aging. Exploring the mechanisms of HGPS aging is not only of great significance for revealing the pathogenesis of HGPS and developing new treatment strategies, but will also provide a new theoretical basis for the study of normal aging.

[0003] Studies have found many similarities between cellular senescence in HGPS patients and physiological cellular senescence, such as the aberrant LMNA splicing leading to low levels of progerin accumulation in physiologically senescent cells. Therefore, HGPS and physiological aging may share the same signaling pathways. Exploring the mechanisms of cellular senescence in HGPS is not only crucial for elucidating its pathogenesis and developing new treatment strategies, but will also provide a new theoretical foundation for research on normal aging.

[0004] Mianserin (MIA, CAS No.: 21535-47-7) is a tetracyclic antidepressant. Mianserin is a norepinephrine and specific 5-HT receptor antagonist (NaSSA), blocking both autoreceptors and heteroreceptors of norepinephrine α2, increasing the concentration of norepinephrine and 5-HT in the synaptic cleft, thus exerting antidepressant, anti-anxiety, and sleep-promoting effects. It also selectively blocks 5-HT2 and 5-HT3 receptors, reducing side effects such as sexual dysfunction and nausea and vomiting. Simultaneously, it blocks H1 receptors, exhibiting a sedative effect. Mianserin can be used as a first-line drug for treating geriatric depression. Its outstanding advantages are fewer adverse reactions, lower toxicity in cases of overdose, and a high safety profile. It is generally considered suitable for outpatients at high risk of overdose, patients with cardiovascular disease, and elderly patients. Summary of the Invention

[0005] The technical problem to be solved by this invention is how to treat aging.

[0006] To address the aforementioned technical problems, the present invention first provides any of the following applications of Mianselin: 1. Its application in the preparation of anti-aging products, or its application in delaying aging; 2. Application in the preparation of products for treating aging, or application in the treatment of aging.

[0007] In the above applications, the aging can be pathological aging or physiological aging.

[0008] Specifically, the physiological aging mentioned above can be natural aging.

[0009] The pathological aging mentioned can be premature aging, such as progeria in children.

[0010] In the above applications, aging can refer to the aging of a human or animal individual, organ, tissue, or cell.

[0011] Furthermore, the animal may be a mammal.

[0012] In the above applications, the delay or treatment of aging can be reflected in improving the external morphology of aging individuals, slowing down the weight loss of aging individuals, improving the mobility of aging individuals, enhancing the grip strength of aging individuals, improving the function of aging organs, reducing the degree of fibrosis in aging tissues, alleviating osteoporosis, and / or improving the content of aging-related factors or inflammatory factors in aging cells.

[0013] Furthermore, the improvement of the external morphology of aging individuals can be reflected in the improvement of their body shape, including size, hair, degree of rickets, etc.

[0014] The aging tissue may be aging heart tissue and / or muscle tissue.

[0015] The aging organ may be an aging liver. Improving the function of the aging organ may be reflected in improving the levels of aspartate aminotransferase (AST) and / or alanine aminotransferase (ALT).

[0016] The aging-related factors may be γH2AX, Cyclin A2, Lamin B1, or PARP1. The inflammatory factors may be IL6 or IL8.

[0017] The present invention also provides a medicament for delaying or treating aging, the medicament containing (or having mianserin as its active ingredient).

[0018] This invention also provides the application of mianserin in the preparation of at least one of the following models: X1) Prepare animal models for improving the external morphology of aging individuals; X2) To prepare an animal model of slowed weight loss in aging individuals; X3) Prepare animal models for improved motor function in aging individuals; X4) To prepare an animal model of enhanced grip strength in aging individuals; X5) Prepare animal organ, tissue, or cell models for improved aging; X6) Prepare animal tissue models with reduced fibrosis in aging tissues; X7) Prepare animal tissue models for osteoporosis remission; X8) Prepare animal cell models that improve aging-related factors or inflammatory factors.

[0019] This invention discovers that mianserin can be used as a novel drug for the treatment or delay of aging: at the cellular level, mianserin can alleviate the aging of HGPS cells and skin fibroblasts in the elderly; at the individual level, mianserin can improve the external morphology of prematurely aging animals, increase their body weight, improve their motor skills and rickets, increase their grip strength and food intake, and improve the degree of fibrosis in the heart and muscle tissue. This demonstrates that mianserin, as an "old drug," can be "repurposed" for the treatment of aging.

[0020] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way. Attached Figure Description

[0021] Figure 1The role of mianserin in HGPS model cell senescence. A: Western blot was used to detect the expression of progerin, cyclin A2, lamin B1, IL6, IL8, γH2AX, PARP1, and S100A6 proteins in CRL-1474 cells overexpressing progerin treated with mianserin, using β-actin as an internal control. The antibodies used were Flag antibody, cyclin A2 antibody, lamin B1 antibody, IL6 antibody, IL8 antibody, γH2AX antibody, PARP1 antibody, S100A6 antibody, and β-actin antibody. "+" indicates the addition of the substance, and "-" indicates its absence. B and C: Immunofluorescence was used to detect and statistically analyze the expression of the cell proliferation marker Ki67 in HGPS model cells after mianserin treatment. The antibody used was Ki67 antibody. D: SA-β-gal staining was used to detect and statistically analyze the role of mianserin in HGPS model cell senescence. E and F: Immunofluorescence assays were performed to detect the expression of the DNA damage marker γH2AX in HGPS model cells after mianserin treatment, and the number of γH2AX foci in the cell nucleus was counted. The antibody used was γH2AX antibody. Vector-Ctrl represents cells transfected with the control virus, Flag-progerin represents HGPS model cells transfected with progerin lentivirus, and Flag-progerin+MIA represents HGPS model cells treated with mianserin.

[0022] Figure 2 Different concentrations of mianserin have the effect on Lmna G609G / G609G The effects of premature aging on the external morphology of mice. A: Lmna G609G / G609G Premature aging mice were treated with intraperitoneal injection of 10 mg / kg micaceline or an equal volume of 0.9% saline (administered every other day) 4 weeks after birth. External morphological changes were observed in the control and treatment groups after 8 weeks of treatment. B: Lmna G609G / G609G Four weeks after birth, premature aging mice were treated with intraperitoneal injection of 20 mg / kg mirtazapine or an equal volume of 0.9% saline (administered every other day). External morphological changes were observed in the control and treatment groups (wild-type mice). Lmna + / + The "-" in the text indicates that no substance was injected; Lmna G609G / G609G In premature aging mice, "-" indicates injection of saline solution, and "+" indicates injection of mianserin.

[0023] Figure 3 Mianselin Lmna G609G / G609GThe effect on body weight in prematurely aging mice. A: Lmna G609G / G609G Four weeks after birth, premature aging mice were treated with intraperitoneal injections of 10 mg / kg micaceline or an equal volume of 0.9% saline (administered every other day). After 8 weeks of treatment, the weight changes of the control and treatment groups were compared, with n=21, 22, and 20 mice in each group. Lmna G609G / G609G Premature aging mice and the 10 mg / kg mianserin treatment group Lmna G609G / G609G Weight changes in premature aging mice at 4 weeks of age (before treatment) and 12 weeks of age (8 weeks after treatment). C: Lmna G609G / G609G Premature aging mice were treated with intraperitoneal injections of 20 mg / kg micaceline or an equal volume of 0.9% saline (administered every other day) 4 weeks after birth. After 8 weeks of treatment, the weight changes of the control and treatment groups were compared, with n=15, 15, and 13 mice in each group. D: Lmna G609G / G609G Premature aging mice and treatment with 20 mg / kg mianserin Lmna G609G / G609G Weight changes in premature aging mice at 4 weeks of age (before treatment) and 12 weeks of age (8 weeks after treatment).

[0024] Figure 4 Different concentrations of mianserin inhibited Lmna G609G / G609G Effects of premature aging on motor function in mice. A: Treatment with 10 mg / kg mianserin (administered every other day). Lmna G609G / G609G Premature aging mice were used, and their motor abilities were assessed and statistically analyzed after 8 weeks. B: Treatment with 20 mg / kg mianserin (administered every other day). Lmna G609G / G609G Premature aging mice were used, and their motor abilities were assessed and statistically analyzed after 8 weeks.

[0025] Figure 5 10 mg / kg of mirtazapine improves Lmna G609G / G609G The degree of rickets in premature aging mice.

[0026] Figure 6 Mianselin Lmna G609G / G609G Effects of Mianserin on cardiac and muscle tissue fibrosis in premature aging mice. A, C: Treatment with Mianserin (administered every other day). Lmna G609G / G609G Premature aging mice were perfused after 8 weeks, and heart and muscle tissues were collected for Masson's staining. The degree of tissue fibrosis was measured using ImageJ. Each group of mice had n=5 mice. B and D represent the quantitative results of the degree of tissue fibrosis.

[0027] Figure 7 Mianselin Lmna G609G / G609G Effects of Mianserin on grip strength in prematurely aging mice. Treatment with Mianserin (administered every other day). Lmna G609G / G609G Eight weeks after the premature aging mice were induced, their grip strength was tested using a grip strength tester. Each group of mice had n=6 mice.

[0028] Figure 8 Mianselin Lmna G609G / G609G Effects of Mianserin on Liver Function in Prematurely Aging Mice. A: Treatment with Mianserin (administered every other day). Lmna G609G / G609G Premature aging mice were treated with AST expression in their serum after 8 weeks, with n=8, 6, and 8 mice in each group. B: Mianserin (administered every other day) was used for treatment. Lmna G609G / G609G In premature aging mice, ALT expression in serum was measured after 8 weeks, with n=8, 6, and 8 mice in each group.

[0029] Figure 9 The effect of mianserin in skin fibroblasts of an 88-year-old patient. Skin fibroblasts derived from an 88-year-old patient were treated with mianserin, and the expression of proteins such as Cyclin A2, Lamin B1, and PARP1 was detected by Western blot after 4 days.

[0030] Figure 10 Mianserin improves the external morphology of naturally aging C57BL / 6 mice. A: 20-month-old naturally aging C57BL / 6 mice were treated with 10 mg / kg mianserin, and their external morphology was observed after 4 months. B: The fur of 20-month-old naturally aging C57BL / 6 mice was evaluated and scored 4 months after administration.

[0031] Figure 11 Effects of mianserin on the skeleton of C57BL / 6 naturally aging mice. Twenty-month-old C57BL / 6 naturally aging mice were treated with 10 mg / kg mianserin (administered every other day), and the skeletons of the mice were examined using Micro-CT after 4 months.

[0032] Figure 12 Effects of mianserin on body weight in naturally aging C57BL / 6 mice. A: Mianserin was used to treat 20-month-old naturally aging C57BL / 6 mice (administered every other day), and the body weight of the mice was monitored. B: After administration of mianserin, the body weight of naturally aging C57BL / 6 mice at 110 weeks of age was significantly higher than that of the control group.

[0033] Figure 13Effects of mianserin on locomotor function in naturally aging C57BL / 6 mice. AB: 20-month-old naturally aging C57BL / 6 mice were treated with 10 mg / kg mianserin (administered every other day). After 4 months, locomotor function was assessed using an open field test.

[0034] Figure 14 Effects of Mianserin on Liver Function in Naturally Aged C57BL / 6 Mice. A: 20-month-old naturally aged C57BL / 6 mice were treated with Mianserin (administered every other day). After 4 months, the expression of AST in the mouse serum was measured. B: 20-month-old naturally aged C57BL / 6 mice were treated. After 4 months, the expression of ALT in the mouse serum was measured. Each group had n=3 mice.

[0035] Figures 10 to 14 In this context, C57BL represents the C57BL / 6 mouse. Detailed Implementation

[0036] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials, reagents, instruments, etc., used in the following examples are all commercially available. All quantitative experiments in the following examples were performed in at least three replicates.

[0037] The following examples use GraphPad Prism software to process the data. The experimental results are expressed as mean ± standard deviation. A two-tailed t-test was used. ns indicates no significant difference, P < 0.05 (*) indicates a significant difference, P < 0.01 (**) indicates a highly significant difference, and P < 0.001 (***) indicates a highly significant difference.

[0038] Premature aging mice in the following examples Lmna G609G / G609G From male and female heterozygotes Lmna G609G / + Mice were mated and screened through genotyping. Among them, Lmna G609G / + The mice were provided by Professor Li Zhanjun's laboratory at the College of Veterinary Medicine, Jilin University, and were of the ICR strain. Lmna G609G / + Mice: Based on the mouse Lmna gene sequence, the sgRNA sequence (Lmna G609G mutation) was obtained using the sgRNADisigner software. An sgRNA expression vector was constructed, and sgRNA and Cas9 mRNA were transcribed in vitro. The resulting mixture was then microinjected into ICR mouse zygotes. Genotyping of the offspring mice was performed after birth to obtain heterozygous mice. Lmna G609G / + Mice.Lmna G609G / + The mice were heterozygous mice obtained by replacing the Lmna gene in the genome of ICR wild-type mice with an allele carrying the c.1827C>T; p.G609G mutation. Lmna G609G / + The biological material described in the literature “NaZhang et al., Unique progerin C-terminal peptide ameliorates Hutchinson-Gilford progeria syndrome phenotype by rescuing BUBR1, Nat Aging. 2023 Feb;3(2):185-201” is available to the public with the consent of Professor Li Zhanjun of the College of Veterinary Medicine, Jilin University. This biological material is only for the purpose of repeating the relevant experiments of this invention and may not be used for other purposes.

[0039] 20-month-old C57BL mice: Zhejiang Vital River Laboratory Animal Technology Co., Ltd.

[0040] Human skin fibroblasts CRL-1474 and human embryonic kidney cells HEK-293T are both products of American Type Culture Collection (ATCC).

[0041] The skin fibroblast cell lines AG05247-88y and AG09602-92y of the elderly are both CCR (Coriell Cell Repository) products.

[0042] The human skin fibroblast cell line CRL-1474, the human embryonic kidney cell line HEK-293T, and the elderly skin fibroblast cell line used in the experiment were all cultured in DMEM medium with 3.7 g / L NaHCO3, 10% fetal bovine serum, 1% Pen / Strep, and pH 7.2. HEK-293T cells were cultured in an incubator at 37°C and 5% CO2. CRL-1474 cells and the elderly skin fibroblast cell line were cultured at 37°C and 5% CO2. 2 ， Incubate in a 5% O2 incubator (Note: The incubator needs to be purged with 99.99% high-purity nitrogen for 24 hours).

[0043] MIA: MCE, HY-B0188A.

[0044] S100A6 antibody: proteintech, 10245-1-AP; Flag antibody: Abmart, M20008; Ki67 antibody: GeneTex GTX16667; γH2AX antibody: Cell Signaling Technology 2916S; IL6 antibody: immunoway, YT5348; IL8 antibody: absin, abs116134; PARP1 antibody: immunoway, YT6210; Lamin B1 antibody: Abcam, ab16048; cyclin A2 antibody: Cell Signaling Technology, 67955S; β-actin antibody: Sigma, A1978; Fluorescent goat anti-rabbit IgG secondary antibody: Invitrogen, A11008; Rabbit anti-sheep IgG secondary antibody: Zhongshan Jinqiao, ZB-2306; Fluorescent goat anti-mouse IgG secondary antibody: Invitrogen, A11029; Goat anti-mouse IgG secondary antibody: Zhongshan Jinqiao, ZF-0313 (Rhodamine); Goat anti-rabbit IgG secondary antibody: Pierce, NC-AP132P-200.

[0045] The pCDH-CMV-3×Flag in the following examples is described in the article “Na Zhang et al., Unique progerin C-terminal peptide ameliorates Hutchinson-Gilford progeria syndrome phenotype by rescuing BUBR1, Nature Aging, Volume 3, February 2023, 185–201”. This biological material is available to the public from the applicant and is only for repeating the relevant experiments of this invention and shall not be used for other purposes.

[0046] Example 1: Mianselin can delay the senescence of HGPS cells. 1. Construction of lentiviral expression plasmids The lentiviral expression plasmid pCDH-CMV-3×Flag-progerin (Hu Qianying et al., Anti-hsa-miR-59 alleviates premature senescence associated with Hutchinson-Gilford progeria syndrome in mice, The EMBO Journal, Volume 42, Issue 1, January 2023) is a recombinant plasmid obtained by inserting progerin into pCDH-CMV-3×Flag. The progerin protein expressed by this plasmid is fused with a Flag tag (denoted as Flag-progerin fusion protein).

[0047] 2. Lentiviral packaging and Mianserin-treated cells 2.1 Packaging of Lentivirals (1) 14-16 hours before transfection, plate HEK-293T cells in 10 cm cell culture dishes. Plate density: so that the cell density reaches about 70-80% at the time of transfection.

[0048] (2) Transfection: On the second day, discard the original culture medium 1 hour in advance and replace it with serum-free and antibiotic-free DMEM culture medium, 5 mL.

[0049] (3) Incubation: Add 500 μL of serum-free and antibiotic-free DMEM medium to a centrifuge tube, 12 μg of core plasmid pCDH-CMV-3×Flag-progerin, 1g of packaging plasmid pMD, 6.5 μg of pRRE, 3.5 μg of VSV-G, 2.5 μg of pRSV-Rsv, and PEI nanomaterial (Sigma, 408727). The total mass of plasmid to the mass ratio of PEI nanomaterial is 4:1. After mixing, let it stand in a clean bench for 15 min, and then slowly add the mixture to a culture dish.

[0050] (4) After 6-8 hours, replace the culture medium with DMEM complete culture medium and continue culturing. On the morning of the third day, continue to change the medium. The virus solution can be collected 48 hours and 72 hours after transfection.

[0051] (5) The virus solution was filtered through a 0.45 μm filter to obtain progerin lentivirus, which was stored at 4°C for later use.

[0052] Following the steps described above, replace the core plasmid with pCDH-CMV-3×Flag, keeping all other steps unchanged, to obtain the control lentivirus.

[0053] 2.2 Viral infection of cells (1) 18-20 h in advance, seed the cells to be infected (human skin fibroblasts CRL-1474) into 10 cm cell culture dishes so that the cell density reaches 60%-70% at the time of infection; (2) On the second day, add 5 mL of filtered virus solution to the cells, then add 10 μL of 1 μg / mL polybrene, and continue to culture the cells for 8-10 h. Change the medium with 10 mL of complete culture medium.

[0054] (3) After 24 hours, cells were collected. Cells obtained from progerin lentivirus were designated as HGPS model cells, and cells obtained from control lentivirus were designated as control cells.

[0055] 2.3 mianselin treatment of cells Mianserin (at a concentration of 10 μM) was added to the cell culture system obtained in step 2.2, and cells were collected after 4 days. The role of mianserin in HGPS cell senescence was detected by Western blot, immunofluorescence, and SA-β-gal staining.

[0056] Experimental results showed that CRL-1474 cells transfected with progerin lentivirus exhibited senescence characteristics in the resulting HGPS model cells, while treatment with mianserin restored the senescence of the HGPS model cells. Figure 1 This indicates that mirtazapine can alleviate the aging of HGPS cells.

[0057] Example 2: Mianselin can delay Lmna G609G / G609G Aging in premature aging mice To further study the effects of Mianselin Lmna G609G / G609G The therapeutic effect on premature aging mice, the inventor was Lmna G609G / G609G Premature aging mice were treated for 8 weeks by intraperitoneal injection of different concentrations of mianserin (10 mg / kg and 20 mg / kg) or 0.9% saline (administered every other day) 4 weeks after birth.

[0058] 1. Mianselin Improvement Lmna G609G / G609G External morphology of premature aging mice Eight weeks after treatment, the external morphology of the mice was observed. Compared with the control group (injected with 0.9% saline), the mice in the 10 mg / kg and 20 mg / kg mianserin treatment groups were larger in size, but smaller than wild-type mice. Lmna + / + Compared to other animals, its size is still relatively small. Figure 2 This result indicates that Mianserin can improve... Lmna G609G / G609GExternal morphology of premature aging mice.

[0059] 2. Mianselin slows down Lmna G609G / G609G Premature aging mice experience weight loss Previous studies have found that HGPS premature aging mice experience a continuous decrease in body weight with age. The inventors treated mice with different concentrations of mianserin... Lmna G609G / G609G The weight changes of premature aging mice were monitored. Results showed that at week 4 after birth, compared with the control group (injected with 0.9% saline), the weight of mice treated with different concentrations of mianserin and the control group were significantly lower. Lmna G609G / G609G There was no significant difference in body weight among the premature aging mice (i.e., there was no significant difference in body weight among the groups before treatment in this experiment). Around 8 weeks after birth, the body weight of mice in both the treated and untreated groups began to decrease, but the decrease was smaller in the mianserin treatment group. Furthermore, at 12 weeks of age, the body weight of mice in both the 10 mg / kg and 20 mg / kg mianserin treatment groups was significantly higher than that of the untreated group. Figure 3 This result indicates that mirtazapine can slow down [the progression of the disease]. Lmna G609G / G609G Decreased body weight in premature aging mice.

[0060] 3. Mianselin Lmna G609G / G609G Effects of premature aging mice on motor function Previous reports have found that decreased motor function is a typical characteristic of HGPS premature aging mice. The inventors tested the effects of different concentrations of mirtazapine on... Lmna G609G / G609G The impact on motor function in prematurely aged mice. Motor function was assessed using an open field test.

[0061] The results showed that after 8 weeks of treatment with 10 mg / kg mirtazapine, Lmna G609G / G609G The motor function of premature aging mice was significantly improved. Figure 4 (Middle A). After 8 weeks of treatment with 20 mg / kg mianserin, the mice's motor function was assessed. Lmna G609G / G609G The motor function of prematurely aged mice was significantly inhibited. Figure 4 (Middle B). This result indicates that miancelin can prolong... Lmna G609G / G609G Motor function in premature aging mice was impaired, but 20 mg / kg mianserin significantly inhibited it. Lmna G609G / G609G Motor function in premature aging mice. Previous studies have found that mianserin has a sedative effect, suggesting that excessively high concentrations of mianserin may affect the motor function of mice. Therefore, subsequent experiments used 10 mg / kg of mianserin to...Lmna G609G / G609G Treating premature aging mice.

[0062] 4. Mianselin Improvement Lmna G609G / G609G Degree of rickets in premature aging mice Furthermore, the inventors treated patients with 10 mg / kg mianserin for 8 weeks... Lmna G609G / G609G Premature aging mice underwent micro-CT scans. The micro-CT detection procedure was as follows: after 8 weeks of treatment with 10 mg / kg mianserin... Lmna G609G / G609G After the premature aging mice were euthanized, their bones were taken for Micro-CT examination, which was performed by the Stomatological Hospital of Jilin University.

[0063] The results showed that after administration Lmna G609G / G609G The degree of rickets on the back of premature aging mice was significantly alleviated. Figure 5 ).

[0064] 5. Mianselin reduces Lmna G609G / G609G Fibrosis of the heart and muscles in premature aging mice In addition, the inventors also used Masson staining to evaluate the effects of 10 mg / kg mianserin treatment. Lmna G609G / G609G The degree of fibrosis in the heart and muscles of premature aging mice was detected using the Masson staining kit (Absin, abs9347).

[0065] The results showed that, compared with the control group, fibrosis in the heart and muscle tissues of mice treated with mianserin was reduced to some extent. Figure 6 ).

[0066] 6. Mianselin Enhancement Lmna G609G / G609G Grip strength of premature aging mice Furthermore, the inventors used a gripping device to treat patients after receiving 10 mg / kg of mirtazapine. Lmna G609G / G609G The grip strength of premature aging mice was tested, and the results showed that after treatment with mianserin... Lmna G609G / G609G The grip strength of premature aging mice increased significantly. Figure 7 ).

[0067] 7. Partial improvement in Mianselin Lmna G609G / G609G Liver function in premature aging mice The above text has established that mianserin can alleviate HGPS aging, and that a dose of 10 mg / kg is most effective. Therefore, long-term use of mianserin is recommended.Lmna G609G / G609G Does premature aging in mice damage liver function? After 8 weeks of treatment with micarin, blood was collected from mice, and the serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were measured using a kit to determine the extent of liver function damage in the mice.

[0068] The results showed that, compared with the untreated group, the serum AST expression and ALT expression of premature aging mice in the mianserin treatment group were significantly decreased, but there was no significant difference between the two groups. This result indicates that mianserin partially improves... Lmna G609G / G609G Liver function in premature aging mice Figure 8 ).

[0069] Example 3: Mianselin can improve the aging phenotype of naturally aging mice. 1. Mianselin can improve the cellular aging phenotype in the elderly. To investigate the role of miancerin in physiological aging, the inventors first treated an 88-year-old skin fibroblast cell line (AG05247-88y) with 10mM miancerin for 4 days, and then detected the expression of proteins such as Cyclin A2, Lamin B1 and PARP1 by Western blot.

[0070] Experimental results showed that mirtazapine upregulated the expression of Cyclin A2, Lamin B1, and PARP1 in cells of an 88-year-old patient, while downregulating the expression of inflammatory factors such as IL6 and IL8. Figure 9 This result indicates that mirtazapine can slow down the aging of cells in an 88-year-old.

[0071] 2. Mianselin can improve the external morphology of naturally aging mice. To investigate the effects of mianserin on naturally aging mice, the inventors intraperitoneally injected 10 mg / kg mianserin or an equal volume of 0.9% saline into 20-month-old male C57BL / 6 mice (administered every other day). Four months later, the external morphology and degree of rickets on the back of the mice were assessed, and their fur was scored. Scoring criteria: 1 point indicates glossy, well-maintained, bright fur with no obvious hair loss; 2 points indicate slight hair loss, revealing some white hairs, unkempt fur, a few small patches on the back, or less than 10% hair loss; 3 points indicate grayish-white fur, dull coat, mild piloerection, a few small patches on the back, or 20%-50% hair loss; 4 points indicate bristles, clumps, dark hair, hair standing on end, large areas of hair loss, or generalized hair loss (not patchy) exceeding 50% hair loss on the back.

[0072] The results showed that mice treated with 0.9% saline were significantly larger and had denser, shinier fur compared to mice treated with 0.9% saline. Figure 10 ).

[0073] 3. Mianselin can improve the degree of rickets in naturally aging mice. The inventors injected 10 mg / kg mianserine or an equal volume of 0.9% saline into 20-month-old male C57BL / 6 mice via intraperitoneal injection. Four months later, the whole skeleton of the mice in the mianserine treatment group and the untreated group was examined by Micro-CT.

[0074] The results showed that mianserin alleviated rickets in the backs of 24-month-old mice. Furthermore, treatment with mianserin significantly reduced osteoporosis in the legs of naturally aging mice. Figure 11 ).

[0075] 4. Mianselin slows down the weight loss in naturally aging C57BL / 6 mice. Previous studies have found that the body weight of both prematurely aging mice and naturally aging mice decreases with age. Furthermore, the inventors have discovered that Mianserin increases... Lmna G609G / G609G The weight of prematurely aging mice. So, does mirtazapine affect the weight of naturally aging mice? The inventors injected 10 mg / kg mirtazapine or an equal volume of 0.9% saline into the intraperitoneal cavity of 20-month-old male C57BL / 6 mice, while simultaneously measuring the mice's weight. The administration continued every other day until the mice died.

[0076] The results showed that the body weight of naturally aging C57BL / 6 mice began to decrease from month 21, but the decrease was smaller in the mianserin treatment group. At week 108, the body weight of mice in the mianserin treatment group was significantly higher than that in the control group. Figure 12 This result indicates that mirtracerin can slow the decline in body weight in naturally aging C57BL / 6 mice.

[0077] 5. Mianselin improves the motor function of naturally aging mice. The inventors have discovered that mianserin can also improve the external morphology of naturally aging C57BL / 6 mice and reduce weight loss. So, can mianserin improve other physiological characteristics and prolong the lifespan of naturally aging C57BL / 6 mice? First, the inventors intraperitoneally injected 20-month-old male C57BL / 6 mice with 10 mg / kg mianserin or an equal volume of 0.9% saline, and examined the effect of mianserin on the mice's motor ability. The results showed that after treatment with mianserin, the motor ability of naturally aging C57BL / 6 mice was significantly enhanced. Figure 13 ).

[0078] 6. Mianserin had no effect on liver function in C57BL / 6 naturally aging mice. In the treatment of naturally aging mice, prolonged administration of medication may affect liver function. Therefore, the inventors tested the liver function of C57BL / 6 naturally aging mice treated with 10 mg / kg mianserin, using AST and ALT levels in the blood as indicators. The results showed that after 4 months of mianserin treatment, there was no damage to liver function in C57BL / 6 naturally aging mice. Figure 14 ).

[0079] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.

Claims

1. The application of Mianserin in the preparation of anti-aging products, or its application in delaying aging.

2. The application of miancelin in the preparation of anti-aging products, or in the treatment of aging.

3. The application according to claim 1 or 2, characterized in that: The aging referred to is either pathological aging or physiological aging.

4. The application according to claim 3, characterized in that: The pathological aging mentioned is premature aging.

5. The application according to any one of claims 1-4, characterized in that: The term "aging" refers to the aging of a person or animal individual, organ, tissue, or cell.

6. The application according to claim 5, characterized in that: The animal in question is a mammal.

7. The application according to any one of claims 1-6, characterized in that: The delay or treatment of aging is reflected in improving the external morphology of aging individuals, slowing down the weight loss of aging individuals, improving the mobility of aging individuals, enhancing the grip strength of aging individuals, improving the function of aging organs, reducing the degree of fibrosis in aging tissues, alleviating osteoporosis, and / or improving the content of aging-related factors or inflammatory factors in aging cells.

8. The application according to claim 7, characterized in that: The improvement of the external morphology of aging individuals is reflected in the improvement of their body shape.

9. A drug for delaying or treating aging, containing mirtracerin.

10. Application of Mianserin in the preparation of any of the following models: X1) Prepare animal models for improving the external morphology of aging individuals; X2) To prepare an animal model of slowed weight loss in aging individuals; X3) Prepare animal models for improved motor function in aging individuals; X4) To prepare an animal model of enhanced grip strength in aging individuals; X5) Prepare animal organ, tissue, or cell models for improved aging; X6) Prepare animal tissue models with reduced fibrosis in aging tissues; X7) Prepare animal tissue models for osteoporosis remission; X8) Prepare animal cell models that improve aging-related factors or inflammatory factors.

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