Traditional Chinese medicine composition for preventing and treating sarcopenia as well as preparation method and application of traditional Chinese medicine composition

Abstract

The traditional Chinese medicine composition is mainly prepared from the following raw material medicines in parts by weight: 1-30 parts of astragalus membranaceus, 1-30 parts of codonopsis pilosula, 1-12 parts of bighead atractylodes rhizome, 1-15 parts of poria cocos, 1-9 parts of herba epimedii and 1-3 parts of liquorice. In addition, the invention further discloses a preparation method and application of the traditional Chinese medicine composition. Pharmacological studies prove that the traditional Chinese medicine composition has the effects of tonifying qi, strengthening the spleen and tonifying the kidney, and can be used for preventing and treating sarcopenia.

Description

Technical Field

[0001] The present invention relates to a traditional Chinese medicine composition, and particularly to a traditional Chinese medicine composition for preventing and treating sarcopenia. In addition, the present invention also relates to a preparation method and application of the traditional Chinese medicine composition. Background Art

[0002] Sarcopenia was first proposed by Rosenberg in 1989 to describe age-related reduction in muscle mass and decline in function. In 2010, the European Working Group on Sarcopenia in Older People (EWGSOP) issued the first expert consensus on this, defining it as a disease mainly characterized by age-related loss of skeletal muscle mass, muscle strength, and / or physical performance decline, and classifying it into primary and secondary types, among which geriatric sarcopenia belongs to the most important primary type. In 2016, the World Health Organization (WHO) officially listed sarcopenia as a disease in the International Classification of Diseases (ICD) [code: ICD (M62.84)]. Currently, China is gradually entering an aging society, and the number of sarcopenia patients is increasing year by year. Since sarcopenia is closely related to limb dysfunction, reduced self-care ability, and a significant increase in fracture risk among the elderly, it has brought a great economic and social burden. In 2019, the Asian Working Group on Sarcopenia (AWGS) found in an epidemiological study of sarcopenia in Asian countries that the prevalence of sarcopenia ranges from 5.5% to 25.7%, with the prevalence in men slightly higher than that in women (5.1% - 21.0% in men and 4.1% - 16.3% in women). In the same year, a Meta-analysis of the epidemiology of sarcopenia in China showed that the prevalence rates of the elderly population in China were 11.0% in the community, 30.0% in hospitals, and 31.0% in nursing homes. Also in the same year, the AWGS updated the original diagnostic guidelines for sarcopenia, making them widely applicable to healthcare and screening in hospitals, research institutions, or communities, and being able to grade the severity of patients based on scoring criteria, further subdivided into possible sarcopenia, sarcopenia, and severe sarcopenia. In 2021, the latest "Expert Consensus on the Diagnosis and Treatment of Sarcopenia in Chinese Elderly" was officially announced, further clarifying the diagnostic and treatment specifications suitable for Chinese elderly sarcopenia patients. Moreover, the data shows that the incidence rate of sarcopenia in Chinese community elderly is 8.9% - 38.8%, and it gradually increases with age, and the incidence rate of the elderly over 80 years old can reach more than 60%. In 2022, the SFDA officially announced the "Guiding Principles for the Design of New Drug Clinical Trials for Sarcopenia", formulating standards for key issues such as its clinical positioning, disease diagnostic criteria, target population selection, exclusion criteria, and efficacy evaluation to facilitate the development of new drugs. Sarcopenia can induce various adverse clinical outcomes, such as falls, hospitalizations, deaths, etc. In the context of an aging society, such a large elderly sarcopenia population has brought a huge health burden and economic burden to families and society. However, so far, the pathogenesis of sarcopenia is still unclear, and there is no effective drug.

[0003] Traditional Chinese medicine (TCM) does not have a specific disease name for sarcopenia. According to its clinical manifestations, it belongs to the category of "flaccidity syndrome" in TCM. TCM had a relatively profound understanding of "flaccidity syndrome" as early as in "Huangdi Neijing", describing it as "flaccidity means weakness and inability to move". Whether in famous medical books such as "Neijing", "Leijing", "Suwen - Treatise on Flaccidity", or "Jingyue Quanshu", many famous TCM physicians unanimously demonstrated that the main pathogenesis of "flaccidity syndrome" is due to external injuries or insufficient kidney qi inherited from parents, resulting in insufficient essence qi, liver and kidney deficiency, lack of nourishment in the acquired stage, and weakness of the spleen qi, thus causing the disease. The deficiency of the congenital and acquired bases of the "kidney" and "spleen" is an important pathogenesis of flaccidity syndrome. Therefore, traditional Chinese medicine compound prescriptions for strengthening the spleen and warming the kidney have the potential to treat sarcopenia.

[0004] In the existing technology, the treatment of sarcopenia mostly adopts methods such as exercise intervention, nutritional support, and muscle electrical stimulation, with mediocre results. There is an urgent need for a traditional Chinese medicine composition to prevent and / or treat sarcopenia. Summary of the Invention

[0005] In order to overcome the deficiencies in the above - mentioned existing technology, in view of the characteristics of sarcopenia with deficiency of both the kidney and spleen and insufficiency of qi and blood, based on the treatment of strengthening the spleen and replenishing qi with Sijunzi Decoction, this invention adds herbs for tonifying the kidney, aiming to tonify both the spleen and kidney and qi and blood. The whole formula replenishes the middle - jiao and benefits qi, and tonifies both the spleen and kidney, providing a traditional Chinese medicine composition for preventing and treating sarcopenia and its preparation method.

[0006] The technical solution adopted by this invention to solve its technical problems is as follows:

[0007] In one aspect of this invention, there is provided a traditional Chinese medicine composition for preventing and treating sarcopenia, comprising the following traditional Chinese medicine raw materials by weight:

[0008] Astragalus membranaceus 1 - 40 parts, Codonopsis pilosula 1 - 40 parts, Atractylodes macrocephala 1 - 20 parts, Poria cocos 1 - 20 parts, Epimedium brevicornu 1 - 20 parts, Glycyrrhiza uralensis 1 - 10 parts.

[0009] The preferred ratio of the above - mentioned raw materials is: Astragalus membranaceus 20 - 30 parts, Codonopsis pilosula 20 - 30 parts, Atractylodes macrocephala 10 - 15 parts, Poria cocos 10 - 15 parts, Epimedium brevicornu 5 - 10 parts, Glycyrrhiza uralensis 2 - 5 parts. The most preferred ratio of the above - mentioned raw materials is: raw Astragalus membranaceus 30 parts, Codonopsis pilosula 30 parts, Atractylodes macrocephala 15 parts, white Poria cocos 15 parts, Epimedium brevicornu 9 parts, roasted Glycyrrhiza uralensis 3 parts.

[0010] Other drugs can also be added based on the above - mentioned traditional Chinese medicine to form various prescriptions and exert different degrees of efficacy.

[0011] As a preferred technical solution of the present invention, in addition to 1-40 parts of Astragalus membranaceus, 1-40 parts of Codonopsis pilosula, 1-20 parts of Atractylodes macrocephala, 1-20 parts of Poria cocos, 1-20 parts of Epimedium brevicornu, and 1-10 parts of Glycyrrhiza uralensis, the raw materials also include one or more of the following drugs: 1-20 parts of Citrus reticulata Blanco and 1-20 parts of Cistanche deserticola. The preferred ratio of the above raw materials is: 30 parts of raw Astragalus membranaceus, 30 parts of Codonopsis pilosula, 15 parts of Atractylodes macrocephala, 15 parts of Poria cocos, 9 parts of Epimedium brevicornu, 3 parts of roasted Glycyrrhiza uralensis, 15 parts of Citrus reticulata Blanco, and 15 parts of Cistanche deserticola.

[0012] The traditional Chinese medicines used in the present invention have definite curative effects, are safe and reliable, and have remarkable curative effects. Among them, Astragalus membranaceus is a qi-tonifying medicine, which is the dried root of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge. Raw Astragalus membranaceus: It is a kind of Astragalus membranaceus, which is a drug that is dried and then remains raw, and is in the form of round or oval slices. Codonopsis pilosula is a qi-tonifying medicine, which is the dried root of Codonopsis pilosula Franch., Codonopsis pilosula Nannf. or Codonopsis tangshen Oliv. Atractylodes macrocephala is a tonifying deficiency medicine, which is the dried rhizome of Atractylodes macrocephala Koidz. Poria cocos is a diuretic and dampness-draining medicine, which is the dried sclerotium of the fungus Poria cocos (Schw.) Wolf. White Poria cocos is the white part after cutting off Red Poria cocos from the Poria cocos block. Epimedium brevicornu is the dried leaf of Epimedium brevicornu Maxim., Epimedium sagittatum (Sieb. et Zucc.) Maxim., Epimedium pubescens Maxim. or Epimedium koreanum Nakai. Glycyrrhiza uralensis is the dried root and rhizome of Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat. or Glycyrrhiza glabra L., and has the effects of replenishing qi and tonifying the middle, relieving spasm and pain, moderating the properties of drugs, eliminating phlegm and relieving cough, and detoxifying. Roasted Glycyrrhiza uralensis belongs to a kind of Glycyrrhiza uralensis, and Glycyrrhiza uralensis belongs to the large category. In traditional Chinese medicine, Glycyrrhiza uralensis is divided into raw Glycyrrhiza uralensis and roasted Glycyrrhiza uralensis. Raw Glycyrrhiza uralensis is cool in nature and is more inclined to clear heat, and its main effects are clearing heat and detoxifying, and moistening the lungs and eliminating phlegm. Roasted Glycyrrhiza uralensis refers to Glycyrrhiza uralensis that has been roasted. Citrus reticulata Blanco is a qi-regulating medicine, which is the dried ripe pericarp of Citrus reticulata Blanco and its cultivated varieties. Cistanche deserticola is a yang-tonifying medicine, which is the dried scaly fleshy stem of Cistanche deserticola Y. C. Ma or Cistanche tubulosa (Schenk) Wight.

[0013] This prescription is based on the Sijunzi Decoction, combined with the qi-tonifying and kidney-tonifying effects of Astragalus membranaceus and Epimedium brevicornu, forming a treatment concept centered on "tonifying qi, strengthening the spleen, and nourishing the kidney". Codonopsis pilosula is used as the sovereign drug, leading to tonify qi and strengthen the spleen. As the core power source of the whole prescription, it has the effects of replenishing middle qi, strengthening the spleen and nourishing the lung, regulating immunity, and anti-fatigue. Astragalus membranaceus is used as the ministerial drug to assist the sovereign drug in enhancing the functions of qi-tonifying and immune enhancement. Epimedium brevicornu is used as the ministerial drug to assist the sovereign drug in tonifying the kidney and strengthening yang, forming a pattern of dual supplementation of "qi-kidney". Atractylodes macrocephala is used as the adjuvant drug, which has the effects of strengthening the spleen and drying dampness, and at the same time regulating the water metabolism in the body. Poria cocos is also used as the adjuvant drug to promote diuresis and percolate dampness, prevent the nourishing drugs from being greasy and obstructing the stomach, and at the same time reconcile the water metabolism in the body. Prepared licorice is used as the guiding drug to reconcile all the drugs, relieve the drastic nature, and enhance the synergy of the whole prescription. This prescription realizes the dynamic balance of "purging while tonifying" by the compatibility of qi-tonifying (Codonopsis pilosula, Astragalus membranaceus) and dampness-dispelling (Atractylodes macrocephala, Poria cocos), avoiding the problem of "stagnation" caused by single supplementation.

[0014] The traditional Chinese medicine composition of the present invention can be prepared into an oral traditional Chinese medicine preparation by the conventional methods of traditional Chinese medicine preparations. For example, the raw medicinal materials are decocted with water, or extracted with an alcohol-water mixture, and then the decoction or extract is concentrated and dried to form a dry extract powder. The extract powder and the medicine are added with appropriate excipients and prepared into an oral traditional Chinese medicine preparation by the conventional pharmaceutical methods of those skilled in the art, including tablets, granules, oral liquids, etc.

[0015] In another aspect of the present invention, there is provided a preparation method of the traditional Chinese medicine composition, including the following steps: weighing the raw medicinal materials according to the said parts by weight, adding 6-12 times the amount of water for decocting, boiling gently for 30 min - 90 min, filtering, adding 4-10 times the amount of water to the medicinal residues, boiling gently for 30 min - 60 min, filtering; combining the two decoction liquids and concentrating them into a concentrated liquid.

[0016] In another aspect of the present invention, there is provided a preparation method of the traditional Chinese medicine composition, including the following steps: weighing the raw medicinal materials according to the said parts by weight, adding 6-12 times the amount of water for decocting, boiling gently for 30 min - 90 min, filtering, adding 4-10 times the amount of water to the medicinal residues, boiling gently for 30 min - 60 min, filtering; combining the two decoction liquids, concentrating, adding appropriate excipients to the extract or the dried extract powder to obtain a granule preparation of the traditional Chinese medicine composition.

[0017] In another aspect of the present invention, there is provided the use of the above traditional Chinese medicine composition in the preparation of a drug for preventing and treating sarcopenia. The traditional Chinese medicine composition of the present invention has better effects in preventing and treating sarcopenia.

[0018] The beneficial effects of the present invention are as follows: According to the physiological and pathological characteristics of sarcopenia, the present invention provides a traditional Chinese medicine formula with remarkable curative effects and no toxic and side effects for the pathological basis of flaccidity syndrome with spleen-kidney deficiency and qi-blood disorder, reflecting the characteristics of traditional Chinese medicine in the dialectical treatment of sarcopenia.

[0019] The concept, specific implementation manners and technical effects of the present invention will be further described below in conjunction with the accompanying drawings to fully understand the purpose, features and effects of the present invention. Description of the Drawings

[0020] Figure 1 It is a graph of the results of a cell activity experiment conducted in Example 9 to detect the effect of the traditional Chinese medicine composition of the present invention on the activity of mouse myoblast C2C12. Specific Implementation Manners

[0021] The present invention will be described below in conjunction with specific embodiments. The technical methods involved, if not specifically described in the present invention, are all technical methods commonly used by those skilled in the art, so they will not be elaborated here.

[0022] Example 1 Preparation of the Pharmaceutical Preparation of the Present Invention

[0023] 30 grams of Astragalus membranaceus, 30 grams of Codonopsis pilosula, 15 grams of Atractylodes macrocephala, 15 grams of Poria cocos, 9 grams of Epimedium brevicornu, 3 grams of roasted licorice.

[0024] Weigh the medicinal materials according to the above prescription, add 10 times the amount of water and decoct for 60 minutes with gentle boiling, filter. Add 8 times the amount of water to the medicinal residues and decoct for 60 minutes with gentle boiling, filter. Combine the two decoction liquids, concentrate under reduced pressure to a relative density of 1.25 - 1.30, use an appropriate amount of dextrin as the base material, spray the concentrated liquid into it, and granulate in a fluidized bed in one step to make 1000 grams, package, 10 grams per package, 2 packages each time, 2 times a day.

[0025] Example 2 Preparation of the Pharmaceutical Preparation of the Present Invention

[0026] 1 gram of Astragalus membranaceus, 1 gram of Codonopsis pilosula, 1 gram of Atractylodes macrocephala, 1 gram of Poria cocos, 1 gram of Epimedium brevicornu, 1 gram of licorice.

[0027] Weigh the medicinal materials according to the above prescription, add 10 times the amount of water and decoct for 60 minutes with gentle boiling, filter. Add 8 times the amount of water to the medicinal residues and decoct for 60 minutes with gentle boiling, filter. Combine the two decoction liquids, concentrate under reduced pressure, dry, add an appropriate amount of lactose, dextrin, and magnesium stearate to the extract powder, mix evenly, granulate to make 1000 grams, package, 10 grams per package, 2 packages each time, 3 times a day.

[0028] Example 3 Preparation of the Pharmaceutical Preparation of the Present Invention

[0029] 40 grams of Astragalus membranaceus, 40 grams of Codonopsis pilosula, 20 grams of Atractylodes macrocephala, 20 grams of Poria cocos, 20 grams of Epimedium brevicornu, 10 grams of licorice.

[0030] Weigh the medicinal materials according to the above prescription, add 10 times the amount of water and decoct. Boil gently for 60 minutes, filter. Add 10 times the amount of water to the residue, boil gently for 30 minutes, filter. Combine the two decoction liquids, concentrate under reduced pressure, dry. Add appropriate amounts of lactose, dextrin, and magnesium stearate to the extract powder, mix evenly, granulate, make 1000 g, package, 10 g per package, 3 times a day, 2 packages each time.

[0031] Example 4 Preparation of the pharmaceutical preparation of the present invention

[0032] Astragalus membranaceus 20 g, Codonopsis pilosula 20 g, Atractylodes macrocephala 10 g, Poria cocos 10 g, Epimedium brevicornu 5 g, Glycyrrhiza uralensis 2 g.

[0033] Weigh the medicinal materials according to the above prescription, add 10 times the amount of water and decoct. Boil gently for 60 minutes, filter. Add 10 times the amount of water to the residue, boil gently for 30 minutes, filter. Combine the two decoction liquids, concentrate under reduced pressure, dry. Add appropriate amounts of lactose, dextrin, and magnesium stearate to the extract powder, mix evenly, granulate, make 1000 g, package, 25 g per package, 3 times a day, 1 package each time.

[0034] Example 5 Preparation of the pharmaceutical preparation of the present invention

[0035] Astragalus membranaceus 30 g, Codonopsis pilosula 30 g, Atractylodes macrocephala 15 g, Poria cocos 15 g, Epimedium brevicornu 10 g, Glycyrrhiza uralensis 5 g.

[0036] Weigh the medicinal materials according to the above prescription, add 10 times the amount of water and decoct. Boil gently for 60 minutes, filter. Add 10 times the amount of water to the residue, boil gently for 30 minutes, filter. Combine the two decoction liquids, concentrate under reduced pressure, dry. Add appropriate amounts of dextrin and magnesium stearate to the extract powder, mix evenly, granulate, make 1000 g, package, 25 g per package, 3 times a day, 1 package each time.

[0037] Example 6 Preparation of the pharmaceutical preparation of the present invention

[0038] Raw Astragalus membranaceus 30 g, Codonopsis pilosula 30 g, Atractylodes macrocephala 15 g, Poria cocos 15 g, Epimedium brevicornu 9 g, Prepared Glycyrrhiza uralensis 3 g, Citrus reticulata Blanco 15 g, Cistanche deserticola 15 g.

[0039] Weigh the medicinal materials according to the above prescription, add 12 times the amount of water and decoct. Boil gently for 90 minutes, filter. Add 4 times the amount of water to the residue, boil gently for 50 minutes, filter. Combine the two decoction liquids, concentrate under reduced pressure, dry. Add appropriate amounts of lactose, dextrin, and magnesium stearate to the extract powder, mix evenly, granulate, make 1000 g, package, 25 g per package, 3 times a day, 1 package each time.

[0040] Example 7 Preparation of the pharmaceutical preparation of the present invention

[0041] Raw Astragalus membranaceus 20 g, Codonopsis pilosula 20 g, Atractylodes macrocephala 10 g, Poria cocos 10 g, Epimedium brevicornu 5 g, Prepared Glycyrrhiza uralensis 2 g, Citrus reticulata Blanco 1 g, Cistanche deserticola 1 g.

[0042] Weigh the medicinal materials according to the above prescription, add 6 times the amount of water and decoct. Bring to a gentle boil for 30 minutes, filter. Add 10 times the amount of water to the residue, bring to a gentle boil for 30 minutes, and filter. Combine the two decoctions, concentrate under reduced pressure, dry. Add an appropriate amount of dextrin and magnesium stearate to the extract powder, mix evenly, granulate, make 1000 g, package, 25 g per pack, take 3 times a day, 1 pack each time.

[0043] Example 8 Preparation of the pharmaceutical preparation of the present invention (decoction)

[0044] Astragalus membranaceus 40 g, Codonopsis pilosula 40 g, Atractylodes macrocephala 20 g, Poria cocos 20 g, Epimedium brevicornu 20 g, Glycyrrhiza uralensis 10 g, Citrus reticulata 20 g, Cistanche deserticola 20 g.

[0045] Extract the above raw materials by water decoction 2 times. For the first decoction, add 1500 ml of water and decoct for 60 minutes, take the remaining juice 150 ml. For the second decoction, add 1000 ml of water and decoct for 45 minutes. For the third decoction, add 1000 ml of water and decoct for 45 minutes, take the remaining juice 150 ml. Take orally twice a day, 150 ml each time.

[0046] Example 9

[0047] The beneficial effects of the present invention are further elaborated through pharmacological studies as follows:

[0048] To detect the effect of the prescription of this medicine on the activity of mouse myoblast C2C12, the following cell activity experiments are carried out.

[0049] 1. Materials and methods:

[0050] 1.1 Experimental materials

[0051] 1.1.1 C2C12 cells were purchased from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences.

[0052] 1.1.2 Main reagents and instruments

[0053] DMEM medium (AJ30740638, HyClone), fetal bovine serum (SH30406.05, HyClone), 0.25% trypsin (P2847398, Adamas life), PBS (23205634, Biosharp). White transparent bottom 96-well cell culture plate (FCP963, Beyotime), 15 mL and 50 mL centrifuge tubes (17122124O, Labselect). Dexamethasone (23030973, Tmsstandard), CellTiter-Lumi TM Plus II luminescence method cell viability detection kit (C0057M, Beyotime). Spark multifunctional microplate reader (Tecan).

[0054] 1.2 Experimental methods

[0055] 1.2.1 Measurement of viability of serum-starved cell model

[0056] Step 1: Inoculate C2C12 myoblasts on cell culture dishes and culture them in proliferation medium (DMEM + 10% fetal bovine serum, 1% penicillin / streptomycin [P / S]), and change the medium at least every 2 days;

[0057] Step 2: Inoculate C2C12 cells (0.5×10 5 cells / well) on 96-well cell culture dishes and culture them overnight in proliferation medium;

[0058] Step 3: Divide C2C12 cells into a normal group, a starvation group, and an experimental group. Among them, the C2C12 cells in the normal group are cultured in proliferation medium, the C2C12 cells in the starvation group are cultured in serum-free medium (DMEM, 1% penicillin / streptomycin [P / S]), and the C2C12 cells in the experimental group are cultured in serum-free medium. Then add the compound extract prepared in Example 1 to the medium at different concentrations (2.5, 5, 10, 25, 50, 75, 100 μg / mL) and culture for 48 hours.

[0059] Step 4: Add 100 μl of CellTiter-Lumi TM Plus II luminescence detection reagent to each well, and detect the chemiluminescence value (RLU) of each well by a multifunctional microplate reader. Each experiment is repeated 3 times, with 6 replicate wells each time.

[0060] 1.2.2 Measurement of viability of dexamethasone-induced cell injury model

[0061] Step 1: Inoculate C2C12 myoblasts on cell culture dishes and culture them in proliferation medium (DMEM + 10% fetal bovine serum, 1% penicillin / streptomycin [P / S]), and change the medium at least every 2 days;

[0062] Step 2: Inoculate C2C12 cells (0.125×10 5 cells / well) on 96-well cell culture dishes and culture them in proliferation medium for 24 hours;

[0063] Step 3: Divide C2C12 cells into a normal group, a dexamethasone group, and an experimental group. The C2C12 cells in the normal group were cultured in a proliferation medium without dexamethasone treatment; the C2C12 cells in the dexamethasone group were treated with 200 μM dexamethasone; the C2C12 cells in the experimental group were treated with 200 μM dexamethasone and different concentrations of the compound extract in Example 1 (2.5, 5, 10, 25, 50, 75, 100 μg / mL) were added to the medium, and cultured for 48 hours.

[0064] Step 4: Add 100 μl of CellTiter-Lumi TM Plus II luminescence detection reagent to each well, and detect the chemiluminescence value of each well through a multifunctional microplate reader. Each experiment was repeated 3 times, with 6 replicate wells each time.

[0065] 2. Statistics and analysis

[0066] The results are as Figure 1 shown. The left figure shows the chemiluminescence values corresponding to the normal group, the starvation group, and the experimental groups at various concentrations under the serum starvation cell model; the right figure shows the chemiluminescence values corresponding to the normal group, the dexamethasone group, and the experimental groups at various concentrations under the dexamethasone-induced cell injury model.

[0067] As Figure 1 shown, the prescription drug has certain protective activities against C2C12 cells damaged by serum starvation and dexamethasone. In the serum starvation cell model, when the concentration of the compound extract in Example 1 is higher than 10 μg / mL, it has an obvious nutritional improvement effect on starved C2C12 cells; in the dexamethasone-induced cell injury model, when the concentration of the compound extract in Example 1 is higher than 5 μg / mL, it has an obvious reversing effect on damaged C2C12 cells.

[0068] Therefore, the compound in Example 1 can significantly improve the survival rate of C2C12 cells under serum starvation and dexamethasone injury, indicating that Example 1 has a good effect on improving sarcopenia.

[0069] Example 10

[0070] In order to investigate the effect of the prescription drug on sarcopenia model mice, the following experiment was carried out.

[0071] 1. Materials

[0072] 1.1 Experimental animals

[0073] Male KM mice, 6 - 8 weeks old, weighing 26 - 30 g, were purchased from Shanghai Model Organisms Center, Inc.

[0074] Five mice were group-housed in each cage and acclimated for 7 days in an environment with a room temperature of 20 - 25°C and a humidity of 30 - 60%, and then the experiment began. During this period, feed and water were provided without discrimination. During the feeding process, the diet, activity, hair color, urine and feces, mental state, etc. of the mice were observed at any time, and the body weight was measured at the same time every day. All animals were handled in accordance with the "Regulations on the Administration of Laboratory Animals" formulated by the National Science and Technology Commission.

[0075] 1.2 Experimental drugs

[0076] Dexamethasone sodium phosphate injection was purchased from Shanghai Pharmaceutical Group Co., Ltd. and used as the model group.

[0077] The prescription drugs were prepared from the following compound ratios, specifically:

[0078] Compound 1 (same prescription as in Example 1): 30 g of Astragalus membranaceus, 30 g of Codonopsis pilosula, 15 g of Atractylodes macrocephala, 15 g of Poria cocos, 9 g of Epimedium brevicornu, 3 g of roasted Glycyrrhiza uralensis;

[0079] Compound 2 (prescription of Shiquandabu Decoction): 12 g of Codonopsis pilosula, 12 g of Atractylodes macrocephala, 12 g of Poria cocos, 6 g of roasted Glycyrrhiza uralensis, 12 g of Rehmannia glutinosa, 10 g of Angelica sinensis, 12 g of Paeonia lactiflora, 6 g of Ligusticum wallichii, 10 g of Astragalus membranaceus, 3 g of Cinnamomum cassia;

[0080] Compound 3 (same prescription as in Example 6): 30 g of Astragalus membranaceus, 30 g of Codonopsis pilosula, 15 g of Atractylodes macrocephala, 15 g of Poria cocos, 9 g of Epimedium brevicornu, 3 g of roasted Glycyrrhiza uralensis, 15 g of Citrus reticulata Blanco, 15 g of Cistanche deserticola;

[0081] Compound 4 (same prescription as in Example 2): 1 g of Astragalus membranaceus, 1 g of Codonopsis pilosula, 1 g of Atractylodes macrocephala, 1 g of Poria cocos, 1 g of Epimedium brevicornu, 1 g of Glycyrrhiza uralensis;

[0082] Compound 5 (same prescription as in Example 3): 40 g of Astragalus membranaceus, 40 g of Codonopsis pilosula, 20 g of Atractylodes macrocephala, 20 g of Poria cocos, 20 g of Epimedium brevicornu, 10 g of Glycyrrhiza uralensis.

[0083] 1.3 Experimental instruments

[0084] Dual-energy X-ray absorptiometer (iNSiGHT VET DXA, OsteoSys, KOREAN); YLS-13A type muscle strength tester, Jinan Yiyan Technology Co., Ltd.

[0085] 2. Methods

[0086] 2.1 Model establishment

[0087] After the above-mentioned male KM mice were adaptively fed, they were randomly grouped, and their weights were measured and numbered. The negative control group (control, blank) was given normal saline, and the sarcopenia model group was given dexamethasone sodium phosphate injection. The administration method was subcutaneous injection. The modeling and administration process lasted for 12 days (2 weeks) (20 mg / kg / day). During this period, food and water were supplied without difference, and the bedding was changed twice a week to keep the living environment clean and hygienic.

[0088] 2.2 Grouping and Administration

[0089] After 48 male KM mice were injected with dexamethasone sodium phosphate for 2 weeks, they were randomly divided into a model group, Compound 1 group (4 g / kg), Compound 2 group (4 g / kg), Compound 3 group (4 g / kg), Compound 4 group (4 g / kg), and Compound 5 group (4 g / kg), with 8 mice in each group; 8 KM mice of the same age were used as the negative control group.

[0090] The extracts of each compound were separately dissolved in 0.3% sodium carboxymethylcellulose solution. Each administration group (including: Compound 1 group (4 g / kg), Compound 2 group (4 g / kg), Compound 3 group (4 g / kg), Compound 4 group (4 g / kg), Compound 5 group (4 g / kg)) was given a dose of 4 g / kg by gavage for 6 consecutive weeks, once a day, and the gavage volume was 0.2 mL / 10 g. The negative control group and the model group were given 0.3% sodium methylcellulose solution by gavage for 6 consecutive weeks, once a day, and the gavage volume was 0.2 mL / 10 g.

[0091] 2.3 Detection Indexes and Methods

[0092] 2.3.1 Measurement of Muscle Strength in Mice

[0093] The forelimb grip strength of mice was measured using a YLS-13A type muscle strength tester (Jinan Yiyan Technology Co., Ltd., Jinan, China). The mice were lifted and fixed by their tails so that their limbs could grasp a wire mesh. Then, the mice were gently pulled backward by their tails, and their postures were parallel to the table surface until they released the mesh. The peak force exerted by the mice's limbs was recorded in grams (g). It was evaluated every 0, 2, 4, and 6 weeks. Each mouse was tested three times, and the average value was taken. After weighting by body weight, it was used for statistical analysis.

[0094] 2.3.2 Measurement of Body Composition in Mice

[0095] Three days before the mice were sacrificed, dual-energy X-ray absorptiometry (DEXA) was used to scan and measure the body composition, including the body fat content, muscle content, bone mineral content, etc. of the mice. The data were analyzed using the software provided by the manufacturer.

[0096] 2.3.3 Statistical Methods

[0097] All data are presented in the form of mean ± standard deviation (mean±SD). One-way ANOVA was performed on the data, and Tukey post-hoc test was used for multiple comparisons. In the results, P < 0.05 indicates significant differences between the two groups of data, P < 0.01 indicates very significant differences between the two groups of data, and P < 0.001 indicates extremely significant differences between the two groups of data. The statistical software used was Graphpad Prism 10.1.2.

[0098] 3. Results

[0099] 3.1 General Observation of Experimental Animals

[0100] The general condition of the negative control group was good, with smooth and shiny hair, and no obvious signs of restlessness. Diet, water intake, and defecation were normal, body weight increased steadily, and the reaction was sensitive. Before the injection of dexamethasone, the mice in each group were in good condition, with shiny fur and sensitive activities. After the start of dexamethasone injection, the model group injected subcutaneously gradually showed restlessness, easy startle, slow reaction, listlessness, fluffy and dull hair, and a sharp increase in body weight. In some mice, necrosis and automatic shedding occurred at the tip of the tail.

[0101] 3.2 Effects of Each Compound Group on Muscle Strength of Model Mice

[0102] As shown in Table 1, before drug administration, compared with the control group, the muscle strength of other groups decreased significantly (P < 0.001); there was no significant difference in the muscle strength of model mice among groups, indicating that the experimental system was reliable and comparable among groups.

[0103] After 2 weeks of drug administration, compared with the control group, the muscle strength of the model group decreased significantly (P < 0.001); compared with the model group, the muscle strength of Compound 1 group increased significantly (P < 0.05), and the muscle strength of Compound 2 group, Compound 3 group, Compound 4 group, and Compound 5 group had an upward trend, but there was no significant difference compared with the model group;

[0104] After 4 weeks of drug administration, compared with the control group, the muscle strength of the model group decreased significantly (P < 0.001); compared with the model group, the muscle strength of Compound 1 group and Compound 2 group increased significantly (P < 0.05), and the muscle strength of Compound 3 group, Compound 4 group, and Compound 5 group had an upward trend, but there was no significant difference compared with the model group;

[0105] After 6 weeks of administration, compared with the control group, the muscle strength of the model group decreased significantly (P < 0.001); compared with the model group, the recovery effects of muscle strength in Compound 1 group and Compound 3 group were most significantly improved (P < 0.001), with the mean values being 6.23 ± 0.57 and 6.01 ± 0.52 respectively. The recovery of muscle strength in Compound 4 group and Compound 5 group was also significantly improved (P < 0.01), with the mean values being 5.43 ± 0.45 and 5.47 ± 0.48 respectively. The recovery of muscle strength in Compound 2 group was also improved with statistical significance (P < 0.05), and the mean value was 5.19 ± 0.66.

[0106] It can be seen from this that Compound 1 group, Compound 3 group, Compound 4 group, and Compound 5 group all had obvious effects on improving the muscle strength of sarcopenia model mice, among which Compound 1 group had the best effect; compared with the effects of other compound groups on improving muscle strength, the improvement effect of Compound 2 group (prescription of Shiquandabutang) was relatively weak.

[0107] Table 1 Effects of each compound group on the muscle strength of model mice (X±SD)

[0108] Before drug administration 2 weeks after drug administration 4 weeks after drug administration 6 weeks after drug administration Control group 5.76±0.43 5.81±0.39 5.94±0.35 6.33±0.51 Model group <![CDATA[3.43±0.37 *** > <![CDATA[3.57±0.53 *** > <![CDATA[3.66±0.49 *** > <![CDATA[3.93±0.60 *** > Compound 1 group <![CDATA[3.24±0.33 *** > <![CDATA[4.38±0.32 # > <![CDATA[4.71±0.67 # > <![CDATA[6.23±0.57 ### <!-- 7 -->]]> Compound 2 group <![CDATA[3.11±0.33 *** > 3.66±0.25 4.11±0.47 <![CDATA[5.19±0.66 # > Compound 3 group <![CDATA[3.60±0.47 *** > 4.33±0.25 <![CDATA[4.65±0.43 # > <![CDATA[6.01±0.52 ### > Compound 4 group <![CDATA[3.04±0.32 *** > 3.87±0.42 4.58±0.49 <![CDATA[5.43±0.45 ## > Compound 5 group <![CDATA[3.25±0.57 *** > 3.98±0.51 4.30±0.35 <![CDATA[5.47±0.48 ## >

[0109] Note: Compared with the control group, ***P < 0.001 in the model group; compared with the model group, #P < 0.05, ##P < 0.01 in the administration group,

[0110] P < 0.001.

[0111] 3.3 Effects of each compound group on the body fat content and muscle content of sarcopenia model mice

[0112] The results after 6 weeks of administration are shown in Table 2. Compared with the control group, the body fat content of the model group increased significantly (P < 0.001); compared with the model group, the body fat content in Compound 1 group, Compound 3 group, Compound 4 group, and Compound 5 group decreased significantly (P < 0.001), among which the improvement of body fat content in Compound 1 group was the most obvious, and the body fat content was 6.89% ± 1.39%; compared with the model group, the decrease in body fat content in Compound 2 group had statistical significance (P < 0.01).

[0113] Compared with the control group, the muscle content of the model group decreased significantly (P < 0.001); compared with the model group, the muscle content in Compound 1 group increased most significantly, with a statistical difference of P < 0.001, and the muscle content ratio was 91.17% ± 2.93%; compared with the model group, the statistical differences in the increase of muscle content in Compound 3 group and Compound 4 group were P < 0.01; compared with the model group, the statistical differences in the increase of muscle content in Compound 2 group and Compound 5 group were P < 0.05.

[0114] It can be seen from this that Compound Group 1, Compound Group 3, Compound Group 4, and Compound Group 5 all have obvious effects on improving the body fat content and muscle content of sarcopenia model mice, among which Compound Group 1 has the best effect; compared with the improvement effects of other compound groups, the improvement effect of Compound Group 2 (the prescription of Shiquandabu Decoction) is relatively weak.

[0115] Table 2 Effects of each compound group on body fat content and muscle content of sarcopenia model mice (X±SD)

[0116] Body fat content (100%) Muscle content (%) Control group 4.34±1.85 91.74±3.18 Model group <![CDATA[12.67±1.09 *** > <![CDATA[83.88±2.30 *** > Compound 1 group <![CDATA[6.89±1.39 ### > <![CDATA[91.17±2.93 ### > Compound 2 group <![CDATA[9.07±1.77 ## > <![CDATA[87.95±2.50 # > Compound 3 group <![CDATA[7.65±2.26 ### > <![CDATA[89.36±2.22 ## > Compound 4 group <![CDATA[7.65±1.64 ### > <![CDATA[88.64±2.23 ## > Compound 5 group <![CDATA[8.18±1.46 ### > <![CDATA[88.23±2.56 # >

[0117] Note: Compared with the control group, *P<0.05, **P<0.01 in the model group; compared with the model group, #P<0.05, ##P<0.01, P<0.001 in the administration group.

[0118] In summary, by verifying that the compounds of each embodiment have obvious improvements on the muscle strength, body fat content, and muscle content of sarcopenia mice, it is proved that the traditional Chinese medicine composition of the present invention indeed has better technical effects in preventing and treating sarcopenia. At the same time, by comparing the compound of the present invention with the compound of Shiquandabu Decoction, it is proved that the traditional Chinese medicine composition of the present invention has a better effect than Shiquandabu Decoction in preventing and treating sarcopenia.

[0119] The preferred specific embodiments of the present invention have been described in detail above. It should be understood that those of ordinary skill in the art can make many modifications and variations based on the concept of the present invention without creative labor. Therefore, all technical solutions that can be obtained by those skilled in the art in the technical field of the present invention through logical analysis, reasoning, or limited experiments based on the concept of the present invention on the basis of the prior art should be within the protection scope determined by the claims.

Claims

1. A Chinese medicine composition for preventing and treating sarcopenia, characterized in that: The invention is mainly prepared from the following raw materials in parts by weight: 1-40 parts of astragalus, 1-40 parts of codonopsis, 1-20 parts of atractylodes, 1-20 parts of tuckahoe, 1-20 parts of epimedium and 1-10 parts of liquorice.

2. The Chinese medicine composition according to claim 1, characterized in that The invention is mainly prepared from the following raw materials in parts by weight: 20-30 parts of astragalus, 20-30 parts of codonopsis, 10-15 parts of atractylodes, 10-15 parts of tuckahoe, 5-10 parts of epimedium and 2-5 parts of liquorice.

3. The Chinese medicine composition according to claim 2, characterized in that: The medicine is mainly prepared from the following raw materials in parts by weight: 30 parts of raw astragalus, 30 parts of codonopsis, 15 parts of atractylodes, 15 parts of white tuckahoe, 9 parts of epimedium and 3 parts of roasted liquorice.

4. The Chinese medicine composition according to claim 1, characterized in that: The raw materials also include the following medicines: 1-20 parts of dried tangerine peel and 1-20 parts of Cistanche deserticola.

5. The Chinese medicine composition according to claim 4, characterized in that: The invention is mainly prepared from the following raw materials in parts by weight: 30 parts of raw astragalus, 30 parts of codonopsis pilosula, 15 parts of atractylodes macrocephala, 15 parts of white tuckahoe, 9 parts of epimedium, 3 parts of roasted licorice, 15 parts of tangerine peel and 15 parts of cistanche deserticola.

6. The Chinese medicine composition according to any one of claims 1 to 5, characterized in that: The Chinese medicine composition is prepared into a Chinese medicine oral preparation.

7. The method for preparing the Chinese medicine composition according to any one of claims 1 to 6, characterized in that: The method comprises the following steps: weighing the raw medicine according to the weight portion, adding 6-12 times of water for decoction, slightly boiling for 30-90 minutes, filtering, adding 4-10 times of water for the medicine residue, slightly boiling for 30-60 minutes, filtering; combining the two decoctions, and concentrating them into a concentrated solution.

8. The method for preparing the Chinese medicine composition according to any one of claims 1 to 6, characterized in that: The method comprises the following steps: weighing raw materials according to the weight portion, adding 6-12 times of water for decoction, slightly boiling for 30-90 minutes, filtering, adding 4-10 times of water for the residue, slightly boiling for 30-60 minutes, filtering; combining the two decoctions, concentrating, adding appropriate amount of auxiliary materials to the extract or the dried extract powder to prepare a granular preparation of the Chinese medicine composition.

9. Use of the Chinese medicine composition according to any one of claims 1 to 6 in the preparation of a medicament for preventing and treating sarcopenia.

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