Application of Anshenbunao liquid in the preparation of anti-aging drugs

Abstract

The present invention provides an application of an Anshen Bu Nao liquid in the preparation of a drug for treating anti-aging, and relates to the technical field of Chinese medicine compositions. In the process of studying the Anshen Bu Nao liquid, the present invention found that the Anshen Bu Nao liquid can significantly improve the cognitive recognition ability and learning and memory ability of rats, increase the number of neurons in diseased rats, and improve neuronal damage and neuroinflammation, indicating that the Anshen Bu Nao liquid of the present invention has a significant anti-aging effect.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine compositions, and in particular relates to the application of anesthesia-shenbunao liquid in the preparation of anti-aging drugs. Background Art

[0002] As the aging trend continues to intensify, how to delay aging has become an important research direction in life sciences. Aging is a natural phenomenon, and anti-aging is the common desire of mankind in ancient and modern times. China has a long history of research on health preservation, anti-aging and longevity. Over thousands of years, it has accumulated rich experience and formed unique theories and methods.

[0003] An important feature of aging is the decline of brain function and the decline of cognition, memory and intelligence. Severe cases of this decline can cause Alzheimer's disease. In recent years, the incidence of vascular dementia, Alzheimer's disease and mixed dementia has shown a significant upward trend. Therefore, the research on drugs that can strengthen the brain, promote intelligence, enhance memory function, prevent and treat Alzheimer's disease has attracted much attention from countries around the world, and the research progress is also relatively fast. There are currently many theories about the mechanism of aging, among which the TCM aging theory mainly includes the kidney deficiency and blood aging theory, the kidney deficiency aging theory, the spleen deficiency aging theory, the body fluid deficiency aging theory, the qi deficiency and blood aging theory, and the spleen and kidney deficiency and gastrointestinal stagnation aging theory.

[0004] A variety of anti-aging drugs have been reported for different aging mechanisms:

[0005] For example, Chinese patent CN113208117A discloses a composition containing NMN and taurine, which contains the following active ingredients: 4-15 parts by weight of nicotinamide mononucleotide (NMN), 0.2-2 parts by weight of nicotinamide, 4-15 parts by weight of taurine, 4-15 parts by weight of vitamin C, 4-15 parts by weight of vitamin E, and 4-15 parts by weight of tea polyphenols. The invention also provides that the above composition can be used in food or dietary supplements, and its efficacy is anti-aging, regulating physiological functions, enhancing immunity, and enhancing brain vitality.

[0006] Chinese patent CN111773362A discloses an anti-aging composition, the active ingredients of which are prepared from the following raw materials in parts by weight: 600-800 parts of velvet antler, 150-200 parts of astragalus, 180-240 parts of yam, 200-260 parts of notoginseng, 200-240 parts of mulberry, 200-300 parts of polygonatum, 60-100 parts of wolfberry, and 100-150 parts of ginger. The composition effectively makes up for the fact that traditional Chinese medicine formulas only focus on tonifying the kidney and yang, reuse kidney-tonifying Chinese medicine, and ignore the important role of spleen-tonifying medicine and blood-activating and blood-stasis-removing medicine, and has the effect of improving body aging. The formula has the effects of delaying brain aging, improving brain function, and enhancing body immune function.

[0007] Chinese patent CN106727799A discloses a health care Chinese medicine composition composed of cordyceps, ginseng and Panax notoginseng. After a large number of formulation studies and clinical screening tests, the composition is composed of 20-60 parts of cordyceps, 20-60 parts of ginseng and 20-60 parts of Panax notoginseng. It has significant effects on improving the body's immune regulation, enhancing vitality, anti-aging and adjusting endocrine, beautifying and removing spots, calming the nerves and nourishing the brain.

[0008] However, the existing Chinese medicine compositions cannot achieve the effects of strengthening the brain, promoting intelligence and enhancing memory. Therefore, it is necessary to develop new uses of tranquilizing the mind and nourishing the brain in tranquilizing the mind, nourishing the brain, enhancing memory and anti-aging. Summary of the invention

[0009] Based on the deficiencies in the prior art, the present invention is based on Anshen Bu Nao Liquid, and studies its preparation method. During the preparation process, different solvents are used to extract the Chinese medicinal ingredients to obtain the Anshen Bu Nao Liquid of the present invention. The Anshen Bu Nao Liquid has the effects of calming the mind and nourishing the brain, enhancing memory, and resisting aging.

[0010] The present invention is achieved like this:

[0011] In one aspect, the present invention provides the use of Anshenbunao liquid in the preparation of anti-aging drugs.

[0012] Furthermore, the drug is a drug that can reduce the level of inflammatory factors.

[0013] Preferably, the inflammatory factors include at least one of IL-1β, IL-6, IL-10 and TNF-α.

[0014] Furthermore, the drug is a drug that increases glutathione peroxidase and / or reduces matrix metalloproteinase 1.

[0015] Furthermore, the drug is a drug that reduces matrix metalloproteinase-1.

[0016] Furthermore, the drug is a drug that reduces transforming growth factor-β.

[0017] Furthermore, the drug is a drug that reduces epidermal growth factor.

[0018] Furthermore, the drug is a drug for lowering C-reactive protein.

[0019] Preferably, the drug is a drug that reduces IL-1β, IL-6, IL-10, TNF-α, matrix metalloproteinase 1, transforming growth factor-β, epidermal growth factor and C-reactive protein, and increases glutathione peroxidase.

[0020] The tranquilizing and brain-nourishing liquid is a product prepared by the methods described in Examples 1 and 2 of the prior patent CN1943774A, or is a tranquilizing and brain-nourishing liquid prepared by the following method.

[0021] The preparation method of the described tranquilizing and brain-nourishing liquid is as follows:

[0022] (1) Mixing velvet antler with water, decocting for 4-6 times, with the solid-liquid ratio of each decoction being 3-8 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the decoction after concentration, standing until the wax layer solidifies, and adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, using a gradually decreasing ethanol concentration during the alcohol precipitation process, collecting the extract after the alcohol precipitation, and obtaining a velvet antler water-extracted alcohol precipitate;

[0023] (2) crushing the dried ginger, mixing it with 6-10 times the amount of water, extracting the distillate by steam distillation, the steam distillation time is 5-12 hours, obtaining the dried ginger volatile oil, and recovering the dried ginger residue;

[0024] (3) Mixing the dried ginger residue with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the amount of the dried ginger residue; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to a concentration of 70-85% in the system, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a dried ginger water extract;

[0025] (4) grinding the prepared Polygonum multiflorum, mixing it with water, and decocting it 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the prepared Polygonum multiflorum; filtering after each decocting, combining the decoctions, concentrating, adding ethanol until the concentration of ethanol in the system is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain the prepared Polygonum multiflorum water extract;

[0026] (5) crushing epimedium, mixing with water, and decocting for 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of epimedium; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain an epimedium water extract;

[0027] (6) Grinding licorice, mixing with water, and decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the licorice powder; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a licorice water extract;

[0028] (7) Mixing jujube with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the jujube; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a jujube water extract;

[0029] (8) The extract components and vitamin B prepared in steps (1) to (7) are 1 , benzoic acid, sodium benzoate, ethylparaben and sucrose are mixed to obtain the tranquilizing and brain-tonifying liquid.

[0030] Preferably, the preparation method of pilose antler water-extract alcohol precipitate comprises the steps of: mixing pilose antler with water, decocting for 4-6 times, with the solid-liquid ratio of each decoction being 3-8 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the concentrated decoction, standing until the wax layer solidifies, adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, using a stepwise decreasing ethanol concentration during the alcohol precipitation process, collecting the extract after the alcohol precipitation, and obtaining pilose antler water-extract alcohol precipitate.

[0031] Further preferably, the alcohol precipitation process specifically comprises the steps of:

[0032] S1, adding ethanol to the remaining part after removing the wax layer until the ethanol concentration in the system is 80-85%, filtering after standing, collecting the filtrate, concentrating to remove ethanol, and obtaining pilose antler alcohol precipitate extract 1;

[0033] S2, continue to add ethanol to the antler alcohol precipitate extract 1 until the ethanol concentration in the system is 70-75%, let it stand and then filter, collect the filtrate, and concentrate to remove ethanol to obtain the antler alcohol precipitate extract 2;

[0034] S3. Add ethanol to the antler alcohol precipitate extract 2 until the ethanol concentration in the system is 20-30%, let it stand and then filter, collect the filtrate, concentrate to remove the ethanol, and dry to obtain the antler water-extracted alcohol precipitate.

[0035] More preferably, during the alcohol precipitation process, the ethanol concentration in the system in each step of steps S1 to S3 is 80%, 75% and 25%, respectively.

[0036] Preferably, the preparation method of dried ginger volatile oil comprises the steps of: taking dried ginger and crushing it, mixing it with 6-10 times the amount of water, extracting the distillate by steam distillation, the steam distillation time is 5-12 hours, obtaining dried ginger volatile oil, and recovering the dried ginger residue for the preparation of dried ginger water extract.

[0037] Further preferably, the dried ginger is crushed to a particle size of 50-100 meshes.

[0038] Further preferably, the amount of water added is 8-9 times the amount of water.

[0039] Further preferably, the steam distillation time is 7-8h.

[0040] Preferably, the preparation method of dried ginger water extract comprises the steps of: mixing dried ginger residue with water, decocting for 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the amount of dried ginger residue; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain the dried ginger water extract.

[0041] Further preferably, the preparation method of dried ginger water extract comprises the steps of: mixing dried ginger residue with water, decocting 3 times, each decoction time is 2 hours, and the amount of water added for each decoction is 8 times the amount of dried ginger residue; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the dried ginger water extract.

[0042] Preferably, the preparation method of the prepared Polygonum multiflorum water extract comprises the steps of: taking the prepared Polygonum multiflorum, crushing it, mixing it with water, decocting it 2-5 times, the decocting time for each time is 1-3 hours, and the amount of water added is 4-10 times the weight of the prepared Polygonum multiflorum; filtering after each decoction, combining the decoctions, concentrating them, adding ethanol until the concentration of ethanol in the system is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain the prepared Polygonum multiflorum water extract.

[0043] Further preferably, the preparation method of the water extract of Polygonum multiflorum comprises the steps of: taking processed Polygonum multiflorum, crushing it, mixing it with water, decocting it 3 times, each decocting time is 2 hours, and the amount of water added for each decoction is 8 times the weight of the processed Polygonum multiflorum; filtering after each decoction, combining the decoctions, concentrating them, adding ethanol to the system until the concentration of ethanol is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the water extract of processed Polygonum multiflorum.

[0044] Preferably, the preparation method of the epimedium water extract comprises the steps of: crushing the epimedium, mixing it with water, decocting it 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the epimedium; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain the epimedium water extract.

[0045] Further preferably, the preparation method of the epimedium water extract comprises the steps of: crushing the epimedium, mixing it with water, decocting it 4 times, each decoction time is 2 hours, and the amount of water added for each decoction is 8 times the weight of the epimedium; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 75%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the epimedium water extract.

[0046] Preferably, the preparation method of licorice water extract comprises the steps of: taking licorice, crushing it, mixing it with water, decocting it 2-5 times, the decocting time of each time is 1-3 hours, and the amount of water added is 4-10 times the weight of licorice powder; filtering after each decoction, combining the decoction liquid, concentrating it, adding ethanol to the system until the ethanol concentration is 70-85%, standing it for 40-68 hours, filtering, collecting the filtrate, concentrating and drying it to obtain the licorice water extract.

[0047] Further preferably, the preparation method of licorice water extract comprises the steps of: taking licorice, crushing it, mixing it with water, and decocting it twice: the first decoction is 1 hour, and the second decoction is 2 hours; the amount of water added in the first decoction is 5 times the weight of the licorice powder, and the amount of water added in the second decoction is 8 times the weight of the licorice powder; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the concentration of ethanol is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the licorice water extract.

[0048] More preferably, the dry grass is crushed, and after the crushing, the licorice with a particle size of 50-100 mesh is sieved and boiled.

[0049] Preferably, the method for preparing the jujube water extract comprises the steps of: mixing jujube with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the jujube; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain the jujube water extract.

[0050] Further preferably, the preparation method of the jujube water extract comprises the steps of: mixing jujube with water and decocting for 3 times, each decocting time is 2 hours, and the amount of water added for each decoction is 5 times the weight of the jujube; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the jujube water extract.

[0051] The tranquilizing and brain-nourishing liquid comprises the following components by weight: 1.5-6 parts of pilose antler, 55-70 parts of processed Polygonum multiflorum, 40-55 parts of epimedium, 8-18 parts of dried ginger, 4.8-7.7 parts of licorice, 8-16 parts of jujube, and 0.2-1 parts of vitamin B 1 .

[0052] Preferably, the tranquilizing and brain-tonifying liquid comprises the following components by weight: 2-5 parts of pilose antler, 60-65 parts of processed Polygonum multiflorum, 45-55 parts of epimedium, 10-15 parts of dried ginger, 5-7 parts of liquorice, 10-15 parts of jujube, and 0.3-0.8 parts of vitamin B 1 .

[0053] The weight ratio of the pilose antler, dried ginger and processed Polygonum multiflorum is 1.5-6:8-18:55-70; the weight ratio is preferably 2-5:10-15:60-65, and the weight ratio is further preferably 3:12.5:62.5.

[0054] As a preferred embodiment, the ingredients are as follows: 3 parts of pilose antler, 62.5 parts of processed Polygonum multiflorum, 50 parts of epimedium, 12.5 parts of dried ginger, 6.25 parts of licorice, 12.5 parts of jujube, 0.5 parts of vitamin B 1 .

[0055] The amounts of benzoic acid, sodium benzoate, ethylparaben and sucrose are conventional amounts in the art.

[0056] Preferably, by weight, sucrose is 100-300 parts, benzoic acid is 1-3 parts, ethylparaben is 0.3-2.3 parts, and sodium benzoate is 1-3 parts.

[0057] Compared with the prior art, the present invention has the following beneficial effects:

[0058] 1. The present invention studies its preparation method and obtains a tranquilizing and brain-tonifying liquid by extracting the traditional Chinese medicine components with different solvents during the preparation process. The tranquilizing and brain-tonifying liquid can better improve the cognitive recognition ability and learning and memory ability of rats, increase the number of neurons in diseased rats, and improve the significant effect of neuronal damage and neuroinflammation; the tranquilizing and brain-tonifying liquid of the present invention has a significant anti-aging effect.

[0059] 2. The present invention makes comprehensive use of various extract ingredients, which are reasonably matched and can be efficiently absorbed to exert their effects; at the same time, through specific extracts and extraction methods, the obtained tranquilizing and brain-tonifying liquid has high activity and significant anti-aging effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 Histogram of total movement distance and average speed of rats in each group in the open field test;

[0061] Figure 2 Bar graph of platform latency and platform crossing times of rats in each group in Morris test (#, CON vs AGE, P<0.05);

[0062] Figure 3 Nissl staining of the cerebral cortex and histogram of the number of neurons in each group of rats;

[0063] Figure 4Changes of CRP, TNF-α, IL-1β, IL-6, IL-10, GSH-Px, TGF-β, EGF, and MMP-1 in the cerebral cortex of rats in each group (###, CON vs AGE, P<0.001; ***, AGE vs drug-treated group, P<0.001). DETAILED DESCRIPTION

[0064] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.

[0065] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings as those generally understood by those of ordinary skill in the art to which the present invention belongs.

[0066] The present invention is further described below by way of specific examples. Unless otherwise specified, the various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels.

[0067] In the following embodiments, "part" and "g" have the same meaning, that is, 1 part = 1 g.

[0068] Basic Example: The preparation method of Anshen Bu Nao Liquid is as follows:

[0069] (1) Mixing velvet antler with water, decocting for 4-6 times, with the solid-liquid ratio of each decoction being 3-8 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the decoction after concentration, standing until the wax layer solidifies, and adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, using a gradually decreasing ethanol concentration during the alcohol precipitation process, collecting the extract after the alcohol precipitation, and obtaining a velvet antler water-extracted alcohol precipitate;

[0070] (2) crushing the dried ginger, mixing it with 6-10 times the amount of water, extracting the distillate by steam distillation, the steam distillation time is 5-12 hours, obtaining the dried ginger volatile oil, and recovering the dried ginger residue;

[0071] (3) Mixing the dried ginger residue with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the amount of the dried ginger residue; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to a concentration of 70-85% in the system, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a dried ginger water extract;

[0072] (4) grinding the prepared Polygonum multiflorum, mixing it with water, and decocting it 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the prepared Polygonum multiflorum; filtering after each decocting, combining the decoctions, concentrating, adding ethanol until the concentration of ethanol in the system is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain the prepared Polygonum multiflorum water extract;

[0073] (5) crushing epimedium, mixing with water, and decocting for 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of epimedium; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain an epimedium water extract;

[0074] (6) Grinding licorice, mixing with water, and decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the licorice powder; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a licorice water extract;

[0075] (7) Mixing jujube with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the jujube; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a jujube water extract;

[0076] (8) The extract components and vitamin B prepared in steps (1) to (7) are 1 , benzoic acid, sodium benzoate, ethylparaben and sucrose are mixed to obtain the tranquilizing and brain-tonifying liquid.

[0077] Basic Example 1: Preparation method of velvet antler water-extracted alcohol precipitation

[0078] The method comprises the following steps: mixing velvet antler with water, decocting 4-6 times, wherein the solid-liquid ratio of each decoction is 3-8 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the decoction after concentration, standing until the wax layer solidifies, adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, collecting the extract after the alcohol precipitation, and obtaining a velvet antler water-extracted alcohol precipitate. The alcohol precipitation process specifically comprises the following steps: (1) first adding ethanol to the remaining part after removing the wax layer until the ethanol concentration in the system is 80%, standing and filtering, collecting the filtrate, concentrating and removing the ethanol, and obtaining a velvet antler alcohol precipitation extract 1; (2) continuing to add ethanol to the velvet antler alcohol precipitation extract 1 until the ethanol concentration in the system is 75%, standing and filtering, collecting the filtrate, concentrating and removing the ethanol, and obtaining a velvet antler alcohol precipitation extract 2; (3) adding ethanol to the velvet antler alcohol precipitation extract 2 until the ethanol concentration in the system is 25%, standing and filtering, collecting the filtrate, concentrating and removing the ethanol, and drying to obtain a velvet antler water-extracted alcohol precipitate.

[0079] Basic Example 2: Preparation method of dried ginger volatile oil and dried ginger water extract

[0080] The method comprises the following steps: crushing dried ginger, mixing dried ginger with a particle size of 50-100 meshes with 9 times the amount of water, extracting distillate by steam distillation, the steam distillation time is 8 hours, obtaining dried ginger volatile oil, and recovering dried ginger medicinal residues for preparing dried ginger water extract.

[0081] The preparation method of the dried ginger water extract comprises the steps of: mixing dried ginger residue with water, decocting for 3 times, each decocting time is 2 hours, and the amount of water added for each decocting is 8 times the amount of the dried ginger residue; filtering after each decocting, combining the decoctions, concentrating, adding ethanol until the concentration of ethanol in the system is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the dried ginger water extract.

[0082] Basic Example 3: Preparation method of Polygonum multiflorum water extract

[0083] The method comprises the following steps: taking processed Polygonum multiflorum, crushing it, mixing it with water, decocting it for 3 times, wherein the decocting time for each time is 2 hours, and the amount of water added for each decoction is 8 times of the weight of the processed Polygonum multiflorum; filtering after each decoction, combining the decoction liquid, concentrating it, adding ethanol until the concentration of ethanol in the system is 80%, standing it for 48 hours, filtering it, collecting the filtrate, concentrating and drying it to obtain the processed Polygonum multiflorum water extract.

[0084] Basic Example 4: Preparation method of Epimedium water extract

[0085] The method comprises the following steps: crushing epimedium, mixing with water, decocting for 4 times, each decocting time being 2 hours, and the amount of water added for each decocting being 8 times the weight of epimedium; filtering after each decocting, combining the decoctions, concentrating, adding ethanol until the concentration of ethanol in the system is 75%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain epimedium water extract.

[0086] Basic Example 5: Preparation method of the liquorice aqueous extract

[0087] The method comprises the following steps: crushing and screening licorice, taking 0.2-0.3 mm licorice and mixing it with water, and decocting it twice: decocting it for 1 hour in the first time, adding water in an amount of 5 times the weight of the licorice powder; decocting it for 2 hours in the second time, adding water in an amount of 8 times the weight of the licorice powder; filtering after each decoction, combining the decoctions, concentrating them, adding ethanol until the concentration of ethanol in the system is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain a licorice water extract.

[0088] Basic Example 6: Preparation method of the jujube water extract

[0089] The method comprises the following steps: mixing jujube with water and decocting for 3 times, wherein the decocting time for each time is 2 hours, and the amount of water added for each decoction is 5 times the weight of the jujube; filtering after each decoction, combining the decoction liquid, concentrating, adding ethanol until the concentration of ethanol in the system is 80%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain the jujube water extract.

[0090] Example 1 A kind of tranquilizing and brain-tonifying liquid with anti-aging effect

[0091] The product prepared by the method described in Example 1 of patent CN1943774A.

[0092] Example 2 A kind of tranquilizing and brain-tonifying liquid with anti-aging effect

[0093] The product prepared by the method described in Example 2 of patent CN1943774A.

[0094] Example 3 A kind of tranquilizing and brain-tonifying liquid with anti-aging effect

[0095] In terms of weight, the ingredients include: 5 parts of pilose antler, 15 parts of dried ginger, 60 parts of processed Polygonum multiflorum, 45 parts of epimedium, 5 parts of licorice, 15 parts of jujube, and 0.7 parts of vitamin B 1 , 300 parts of sucrose, 2.5 parts of benzoic acid, 2.0 parts of ethylparaben and 3 parts of sodium benzoate.

[0096] Preparation method: Prepare various extracts and vitamin B according to basic examples 1-6 1 , benzoic acid, sodium benzoate, ethylparaben and sucrose are mixed, and water is added to 1000 ml to obtain the described tranquilizing and brain-tonifying liquid.

[0097] Example 4 A kind of tranquilizing and brain-tonifying liquid with anti-aging effect

[0098] In terms of weight, the ingredients include: 1.5 parts of velvet antler, 8 parts of dried ginger, 55 parts of processed Polygonum multiflorum, 40 parts of epimedium, 4.8 parts of licorice, 8 parts of jujube, and 0.2 parts of vitamin B 1, 100 parts of sucrose, 1 part of benzoic acid, 0.3 parts of ethylparaben and 1 part of sodium benzoate.

[0099] Preparation method: Prepare various extracts and vitamin B according to basic examples 1-6 1 , benzoic acid, sodium benzoate, ethylparaben and sucrose are mixed, and water is added to 1000 ml to obtain the described tranquilizing and brain-tonifying liquid.

[0100] Example 5 A tranquilizing and brain-tonifying liquid with anti-aging effects

[0101] In terms of weight, the ingredients include: 3 parts of velvet antler, 12.5 parts of dried ginger, 62.5 parts of processed Polygonum multiflorum, 50 parts of epimedium, 6.25 parts of licorice, 12.5 parts of jujube, and 0.5 parts of vitamin B 1 , 200 parts of sucrose, 2.2 parts of benzoic acid, 1.8 parts of ethylparaben and 1.5 parts of sodium benzoate.

[0102] Preparation method: Prepare various extracts and vitamin B according to basic examples 1-6 1 , benzoic acid, sodium benzoate, ethylparaben and sucrose are mixed, and water is added to 1000 ml to obtain the described tranquilizing and brain-tonifying liquid.

[0103] Comparative Example 1

[0104] The difference from Example 5 is that 55 parts of Epimedium are replaced by 55 parts of Poria.

[0105] The preparation method of Poria cocos extract is the same as that of Epimedium extract: comprising the steps of: crushing Poria cocos, mixing with water, decocting 4 times, each decocting time is 2 hours, and the amount of water added for each decoction is 8 times the weight of Poria cocos; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 75%, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain Poria cocos water extract.

[0106] The rest is the same as in Example 1, and the product of Comparative Example 1 is obtained.

[0107] Comparative Example 2

[0108] Different from Example 5, the Anshenbunao liquid does not contain the component Polygonum multiflorum.

[0109] Comparative Example 3

[0110] The difference from Example 5 is that the particle size of the dried ginger powder in step (2) is different. In this comparative example, the particle size of the dried ginger powder used in preparing the dried ginger volatile oil is between 10 and 20 meshes. The rest is the same as Example 1.

[0111] Comparative Example 4

[0112] The difference from Example 5 is that the preparation method of Anshenbunao liquid is different, specifically a one-step mixing water extraction method of Chinese medicinal ingredients:

[0113] (1) Same as Example 5;

[0114] (2) crushing the dried ginger, mixing the dried ginger with a mesh size of 50-100 with water, and decocting for 3 times, each decocting time is 2 hours, and the amount of water added for each decocting is 8 times the amount of the dried ginger residue; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to a concentration of 80% in the system, standing for 48 hours, filtering, collecting the filtrate, concentrating and drying to obtain a dried ginger water extract; and recovering the dried ginger residue;

[0115] (3) Mix the crushed dried ginger residue, processed Polygonum multiflorum, epimedium, jujube and liquorice (50-100 mesh) with water and decoct for 3 times, each decoction time is 2 hours, and the amount of water added for each decoction is 8 times the weight of the Chinese medicine; filter after each decoction, combine the decoctions, concentrate, add ethanol to the system until the ethanol concentration is 80%, let stand for 48 hours, filter, collect the filtrate, concentrate and dry to obtain a water extract mixture;

[0116] (4) mixing the extract prepared in steps (1) to (3) with vitamin B 1 , benzoic acid, sodium benzoate, ethylparaben and sucrose are mixed, and water is added to 1000 ml to obtain the described tranquilizing and brain-tonifying liquid.

[0117] Effect experiment

[0118] 1. Purpose of the experiment

[0119] To evaluate the anti-aging pharmacological effects of Anshenbu'nao liquid.

[0120] 2 Experimental Materials

[0121] 2.1 Experimental animals

[0122] 140 male SD rats, weighing (220±20) g, were provided by Beijing Huafukang Biotechnology Co., Ltd. The animals were kept in the Experimental Animal Center of Guangdong Provincial Hospital of Traditional Chinese Medicine at a room temperature of 18-23°C and a relative humidity of 45%-55%, with free access to water and food, and were adapted for 1 week. All animal experiments were conducted in accordance with the guidelines of the Experimental Animal Center of Guangdong Provincial Hospital of Traditional Chinese Medicine and approved by the Animal Ethics Committee of Guangdong Provincial Hospital of Traditional Chinese Medicine (2022016).

[0123] 2.2 Reagents and materials

[0124] The invention prepares the anshenbunao liquid (oral administration by gavage), rapamycin (prepared as 0.05 mg / mL solution for gavage), 0.9% sodium chloride injection (intraperitoneal injection), D-galactose (prepared as 120 mg / mL solution for injection), and isoflurane (inhalation anesthesia).

[0125] 2.3 Experimental instruments

[0126] The animal behavior and pain measurement system (model: Panlabharved, Huaibei Zhenghua Biological Instrument Equipment Co., Ltd.) includes Morris water maze constant temperature swimming pool and open field ZH-ZFT, animal behavior software SMART 3.0 (Panlab, Spain), fully automatic intelligent dehydrator (model: HP300, Dakoway Biotechnology Co., Ltd.), tissue embedding machine (model: HistoStar, Thermo Fisher Scientific (China) Co., Ltd.), fully automatic staining and sealing integrated workstation (model: ST5020, Leica Instrument Co., Ltd., Germany), paraffin sectioner (model: RM2245, Leica Instrument Co., Ltd., Germany), upright fluorescence microscope (model: BX53, Olympus (China) Co., Ltd.), and medical centrifuge (model: DT5-3, Beijing Times Beili Centrifuge Co., Ltd.).

[0127] 3 Experimental methods

[0128] 3.1 Animal grouping and drug administration

[0129] After 3 days of adaptive feeding, 140 SD male rats were randomly divided into a normal control group (CON), an aging model group (AGE), a low-dose group (ASL), a medium-dose group (ASM) and a high-dose group (ASH) of Example 1, Example 2-5 groups, Comparative Example 1-4 groups, and a rapamycin group (RAP), with 10 rats in each group. The normal control group and the model group were given distilled water by gavage at 1 mL / 100 g, the positive control group was given rapamycin by gavage at 0.5 mg / kg / d, the low, medium and high dose groups of Example 1 were given the Anshenbunao liquid prepared in Example 1 by gavage at a dose of 1.05 mL / kg / d, 2.1 mL / kg / d, and 4.2 mL / kg / d, the Example 2-5 group was given the Anshenbunao liquid prepared in Example 2-5 by gavage at a dose of 2.1 mL / kg / d, and the Comparative Example 1-4 group was given the Anshenbunao liquid prepared in Comparative Example 1-4 by gavage at a dose of 2.1 mL / kg / d, and the administration was continued for 8 weeks. Modeling was started 2 weeks after administration. The model group and all the treatment groups were given D-galactose at 120 mg / kg / d by subcutaneous injection, and the normal control group was given an equal amount of saline by subcutaneous injection, and the modeling was continued for 6 weeks.

[0130] 3.2 Open field experiment

[0131] The open field test is used to detect the behavior of animals in unfamiliar environments, including spontaneous activity and exploratory behavior. It is a method to evaluate the autonomous behavior, exploratory behavior and tension of animals in a new environment. The steps are as follows: ① Before the experiment, the animals should adapt to the test environment for 30-60 minutes; ② The experimenter pre-sets the corresponding parameters in the Smart 3.0 software and records the animal's number, date, and status information; ③ After the animal is placed in the test box, the test begins, and the test time is 5 minutes; ④ Each time the experiment is carried out, the animal should be placed in the center of the test box from the same position and direction.

[0132] 3.3 Morris water maze

[0133] Hidden platform test: At the beginning of each day, the mouse is placed in the water facing the pool wall from any of the four quadrants (northeast, northwest, southeast, southwest), and the platform is placed in the southeast quadrant. Each time the mouse swims for a total of 60 seconds to find the hidden platform. If the platform is successfully found, the mouse can get a 10-second rest time on the platform; if the platform is not successfully found within 60 seconds, the experimenter will manually place the mouse on the platform and give it a 10-second rest time. Sometimes the mouse may fall off the platform or jump into the water to continue swimming before the 10-second interval is reached. Once this happens, the mouse is put back on the platform and the time is reset so that the time interval reaches (10 seconds). This ensures that each mouse has equal time to observe and obtain spatial information after each experiment.

[0134] Spatial exploration test: 24 hours after the hidden platform test, the platform was removed. Then, the mouse was placed in the water at the same entry point, and the swimming path of the mouse within 60 seconds was recorded. The mouse's stay time in the original platform quadrant and the number of times it crossed the original platform were recorded. The spatial positioning ability of the test mouse and the changing rules during the spatial exploration process were observed. The original location of the platform was marked with a circular ring on the computer screen, so that the number of times the original platform location was crossed could be recorded.

[0135] 3.4 Histopathology

[0136] The brain and colon immersed in fixative were dehydrated, embedded, dewaxed, sectioned, and stained with HE and Nissl according to standard procedures.

[0137] 3.5 Elisa test

[0138] According to the instructions, the contents of interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 10 (IL-10), tumor necrosis factor-α (TNF-α), C-reactive protein (CRP), glutathione peroxidase (GSH-Px), transforming growth factor-β (TGF-β), epidermal growth factor (EGF), and matrix metalloproteinase 1 (MMP-1) in the rat cerebral cortex were detected.

[0139] 3.6 Statistical analysis

[0140] The data were imported into GraphPad Prism 8 software for graphing and statistical analysis. Welch's t test was used for comparison analysis between two groups. P < 0.05 indicated that the difference was statistically significant.

[0141] 4 Experimental results

[0142] 4.1 Open field experiment

[0143] In the open field test, the total movement distance and average movement speed of the rats in the aging model group were reduced compared with the normal control group. The total movement distance of the rats in the medium dose group of Anshen Bu Nao Liquid of Example 2-5 was greater and the average speed was faster than that of the rats in the model group.

[0144] Compared with the normal control group, the total movement distance of the rats in the model group was significantly shortened. The low, medium and high doses of the Anshenbu'nao solution in Example 1 can increase the total movement distance of the rats. Rapamycin has a stronger effect on increasing the total movement distance of rats than Anshenbu'nao solution.

[0145] The open field test showed that: ① the autonomous movement ability of rats in the D-galactose-induced aging model was significantly reduced; ② both Anshenbu'nao liquid and positive drug (rapamycin) could improve the autonomous movement ability of aging rats; ③ different doses of Anshenbu'nao liquid could improve the autonomous movement ability of aging rats, among which the high-dose group had better efficacy than the low- and medium-dose groups, showing a certain dose-effect relationship.

[0146] 4.2 Morris water maze

[0147] In the Morris water maze, the platform latency of the model group rats was significantly prolonged compared with the normal control group. The platform latency of the rats in the group receiving the medium dose of the Anshenbunao solution of Example 2-5 was shortened.

[0148] Compared with the normal control group, the number of times the model group rats crossed the platform was significantly reduced. Low, medium and high doses of the Anshenbu'nao solution in Example 1 can increase the number of times the platform is crossed. Rapamycin has a stronger effect on increasing the number of times the rats cross the platform than the Anshenbu'nao solution.

[0149] Moriis' experiment showed that: ① The learning and memory abilities of rats in the aging model induced by D-galactose were significantly reduced; ② Anshenbu'nao liquid and positive drugs (rapamycin) could improve the learning and memory abilities of aging rats; ③ Different doses of Anshenbu'nao liquid could improve the learning and memory abilities of aging rats, among which the high-dose group had better efficacy than the low- and medium-dose groups, showing a certain dose-effect relationship.

[0150] 4.3 Histopathology

[0151] From the statistical results, compared with the normal control group, the number of neurons in the cerebral cortex of the aging model group rats decreased. Different types and doses of Anshenbunao liquid can increase the number of neurons in aging rats. The high dose of Anshenbunao liquid in Example 1 has better efficacy than the low and medium doses. The positive drug rapamycin can also increase the number of neurons.

[0152] Histopathology showed that: ① The number of neurons in the cerebral cortex of D-galactose-induced aging model rats decreased; ② Both Anshenbu'nao liquid and positive drug (rapamycin) could increase the number of neurons in aging rats; ③ Different doses of Anshenbu'nao liquid could increase the number of neurons in aging rats, among which the high-dose group had better efficacy than the low- and medium-dose groups, showing a certain dose-effect relationship.

[0153] 4.4 Elisa test

[0154] From the perspective of inflammatory factor indicators, compared with the normal control group, the inflammatory response in the aging model group rats increased significantly, as shown by a significant increase in the content of pro-inflammatory factors CRP, TNF-α, IL-1β, and IL-6, and a significant decrease in the content of anti-inflammatory factor IL-10. Different types and doses of Anshenbu Nao Liquid can reverse the excessive inflammatory response caused by aging. The positive drug rapamycin can also inhibit the inflammatory response in aging rats.

[0155] From the perspective of oxidative stress indicators, compared with the normal control group, the oxidative stress in the aging model group rats increased significantly, as shown by a significant decrease in the enzyme activity of GSH-Px. Different doses of Anshenbu Nao Liquid can reverse the excessive oxidative stress caused by aging. The positive drug rapamycin can also inhibit oxidative stress in aging rats.

[0156] From the perspective of growth factor indicators, compared with the normal control group, the levels of TGF-β, EGF, and MMP-1 in the aging model group rats increased significantly, and an aging-related secretory phenotype appeared. Different doses of Anshenbu Nao Liquid can reverse the excessive TGF-β, EGF, and MMP-1 caused by aging. The positive drug rapamycin can also inhibit the aging-related secretory phenotype in aging rats.

[0157] Table 1 The contents of CRP, TNF-α, IL-1β, IL-6, IL-10, GSH-Px, TGF-β, EGF and MMP-1 in the cerebral cortex of rats in each group ( ### , normal control group vs aging model group, P<0.001; *** , aging model group vs drug administration group, P<0.001)

[0158]

[0159]

[0160]

[0161] Elisa test results show:

[0162] ① In the D-galactose-induced aging model rats, inflammatory responses were overactivated and oxidative stress occurred, and the secretion of growth factors and protease regulators was significantly increased;

[0163] ②Anshenbunao solution and positive drug (rapamycin) can improve the aging-related secretory phenotype;

[0164] ③ Different doses of Anshenbunao solution can increase the number of neurons in aging rats, among which the high-dose group has better efficacy than the low-medium dose group, showing a certain dose-effect relationship;

[0165] ④ The Anshen Bu Nao Liquid prepared in Comparative Examples 1-4 increased the number of neurons in aging rats significantly less than the Anshen Bu Nao Liquid prepared in Example 1-5.

[0166] The above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the same. Although the present invention has been described in detail with reference to the above embodiments, a person skilled in the art can still modify or make equivalent substitutions to the specific implementation schemes of the present invention. Any modifications or equivalent substitutions that do not depart from the spirit and scope of the present invention are within the scope of protection of the claims of the present invention.

Claims

1. Application of Anshenbunao liquid in the preparation of anti-aging drugs; The tranquilizing and brain-nourishing liquid comprises the following ingredients, measured by weight: 1.5-6 parts of pilose antler, 55-70 parts of processed polygonum multiflorum, 40-55 parts of epimedium, 8-18 parts of dried ginger, 4.8-7.7 parts of liquorice, 8-16 parts of jujube, and 0.2-1 parts of vitamin B1; The preparation method of the tranquilizing and brain-tonifying liquid is as follows: (1) Mixing velvet antler with water, decocting for 4-6 times, with the solid-liquid ratio of each decoction being 3-8 g / mL, filtering, and combining the filtrate to obtain a decoction; adding beeswax to the decoction after concentration, standing until the wax layer solidifies, and adding ethanol to the remaining part after removing the wax layer for alcohol precipitation, using a gradually decreasing ethanol concentration during the alcohol precipitation process, collecting the extract after the alcohol precipitation, and obtaining a velvet antler water-extracted alcohol precipitate; The alcohol precipitation process specifically comprises the steps of: S1, adding ethanol to the remaining part after removing the wax layer until the ethanol concentration in the system is 80-85%, filtering after standing, collecting the filtrate, concentrating to remove ethanol, and obtaining pilose antler alcohol precipitate extract 1; S2, continue to add ethanol to the antler alcohol precipitate extract 1 until the ethanol concentration in the system is 70-75%, let it stand and then filter, collect the filtrate, and concentrate to remove ethanol to obtain the antler alcohol precipitate extract 2; S3, add ethanol to the antler alcohol precipitate extract 2 until the ethanol concentration in the system is 20-30%, let it stand and then filter, collect the filtrate, concentrate to remove ethanol, and dry to obtain the antler water-extracted alcohol precipitate; The ethanol concentration in each system of steps S1-S3 is 80%, 75% and 25% respectively; (2) crushing the dried ginger to a particle size of 50-100 mesh, mixing it with 6-10 times the amount of water, and extracting the distillate by steam distillation for 5-12 hours to obtain the dried ginger volatile oil, and recovering the dried ginger residue; (3) Mixing the dried ginger residue with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the amount of the dried ginger residue; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to a concentration of 70-85% in the system, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a dried ginger water extract; (4) grinding the prepared Polygonum multiflorum, mixing it with water, and decocting it 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the prepared Polygonum multiflorum; filtering after each decocting, combining the decoctions, concentrating, adding ethanol until the concentration of ethanol in the system is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain the prepared Polygonum multiflorum water extract; (5) crushing epimedium, mixing with water, and decocting for 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of epimedium; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain an epimedium water extract; (6) Grinding licorice, mixing with water, and decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the licorice powder; filtering after each decoction, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a licorice water extract; (7) Mixing jujube with water, decocting 2-5 times, each decocting time is 1-3 hours, and the amount of water added is 4-10 times the weight of the jujube; filtering after each decocting, combining the decoctions, concentrating, adding ethanol to the system until the ethanol concentration is 70-85%, standing for 40-68 hours, filtering, collecting the filtrate, concentrating and drying to obtain a jujube water extract; (8) Mix the extract components prepared in steps (1) and (3)-(7), vitamin B1, benzoic acid, sodium benzoate, ethylparaben and sucrose to obtain the tranquilizing and brain-tonifying solution.

2. The use according to claim 1, characterized in that: The medicine is a medicine that can reduce the level of inflammatory factors.

3. The use according to claim 2, characterized in that: The inflammatory factors include at least one of IL-1β, IL-6, IL-10 and TNF-α.

4. The use according to claim 1, characterized in that: The drug is a drug for increasing glutathione peroxidase.

5. The use according to claim 1, characterized in that: The drug is a drug for reducing matrix metalloproteinase 1.

6. The use according to claim 1, characterized in that: The medicine is a medicine for reducing transforming growth factor-β.

7. The use according to claim 1, characterized in that: The medicine is a medicine for reducing epidermal growth factor.

8. The use according to claim 1, characterized in that: The drug is a drug for reducing C-reactive protein.

9. The use according to any one of claims 3 to 8, characterized in that: The drug is a drug that reduces IL-1β, IL-6, IL-10, TNF-α, matrix metalloproteinase 1, transforming growth factor-β, epidermal growth factor and C-reactive protein, and increases glutathione peroxidase.

10. The use according to claim 1, wherein the Anshen Bu Nao Liquid can be administered alone or in combination with one or more other therapeutic agents.

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