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

Abstract

The traditional Chinese medicine composition is mainly prepared from the following raw material medicines in parts by weight: 1-30 parts of ginseng, 1-30 parts of pseudo-ginseng, 1-12 parts of Chinese yam, 1-15 parts of poria cocos, 1-9 parts of cinnamon, 1-9 parts of herba epimedii, 1-9 parts of red dates and 1-3 parts of liquorice. In addition, the invention further discloses a preparation method and application of the traditional Chinese medicine composition. Pharmacological research proves that the traditional Chinese medicine composition has the effects of replenishing qi and enriching blood, and tonifying spleen and kidney, and can be used for preventing and treating sarcopenia.

Description

Technical Field

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

[0002] Sarcopenia was first proposed by Rosenberg in 1989 to describe the phenomenon of age-related muscle mass loss and functional decline. In 2010, the European Working Group on Sarcopenia (EWGSOP) released the first expert consensus, defining it as a disease with age-related loss of skeletal muscle mass, muscle strength and / or physical fitness as the main clinical features, and dividing it into two types: primary and secondary. Among them, elderly sarcopenia is the most important primary type. In 2016, the World Health Organization (WHO) International Classification of Diseases (ICD) officially listed sarcopenia as a disease [code is ICD (M62.84)]. At present, my country 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 significantly increased risk of fractures in the elderly, it has brought great economic and social burdens. In 2019, the Asian Working Group on Sarcopenia (AWGS) conducted an epidemiological study on sarcopenia in Asian countries and found that the prevalence of sarcopenia ranged from 5.5% to 25.7%, with a slightly higher prevalence in men than in women (5.1% to 21.0% in men and 4.1% to 16.3% in women). In the same year, a meta-analysis of the epidemiology of sarcopenia in China showed that the prevalence rates in the elderly population in China were 11.0% in the community, 30.0% in hospitals, and 31.0% in nursing homes. In the same year, AWGS updated the original diagnostic guidelines for sarcopenia, making it widely used in medical care and screening in hospitals, research institutions or communities, and able to grade the severity of patients based on scoring criteria, further subdivided into possible sarcopenia, sarcopenia and severe sarcopenia. In 2021, the latest "Sarcopenia in the Elderly in China" was published. The "Expert Consensus on the Diagnosis and Treatment of Sarcopenia" was officially announced, further clarifying the diagnosis and treatment standards suitable for elderly patients with sarcopenia in my country. In addition, the data show that the incidence of sarcopenia in the elderly in the Chinese community is 8.9% to 38.8%, and it gradually increases with age. The incidence rate in the elderly over 80 years old can reach more than 60%. In 2022, SFDA officially announced the "Guidelines for the Design of Clinical Trials of New Drugs for Sarcopenia", and formulated standards for key issues such as its clinical positioning, disease diagnosis 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, hospitalization, and death. In the context of an aging society, such a large group of elderly people with sarcopenia has brought huge health and economic burdens to families and society. However, the pathogenesis of sarcopenia is still unclear and there is no effective drug to date.

[0003] Sarcopenia does not have a disease name in traditional Chinese medicine. According to its clinical manifestations, it belongs to the category of "paralysis" in traditional Chinese medicine. Traditional Chinese medicine had a deep understanding of "paralysis" as early as the Yellow Emperor's Classic of Internal Medicine, describing it as "paralysis means weakness and inability to move". Whether in famous medical books such as the Internal Classic, Classified Classics, Suwen-Paradox, or Jingyue Complete Book, many famous doctors have unanimously demonstrated that the main pathogenesis of "paralysis" is due to external damage or the patient's parents' insufficient kidney qi, which leads to insufficient essence and qi, liver and kidney deficiency, postnatal malnutrition, and spleen deficiency. The innate and acquired deficiency of "kidney" and "spleen" is an important pathogenesis of paralysis. Therefore, Chinese herbal compound prescriptions that strengthen the spleen and warm the kidney have the potential to treat sarcopenia.

[0004] In the prior art, the treatment of sarcopenia mostly adopts exercise intervention, nutritional support and muscle electrical stimulation, etc., but the effect is mediocre. There is an urgent need for a Chinese medicine composition to prevent and / or treat sarcopenia. Summary of the invention

[0005] In order to overcome the deficiencies in the above-mentioned prior art, the present invention aims at the characteristics of sarcopenia such as deficiency of both kidney and spleen and insufficient qi and blood, and adopts a prescription composed of Chinese medicines for tonifying both spleen and kidney and qi and blood, to provide a Chinese medicine composition for preventing and treating sarcopenia that can tonify the middle and replenish qi, and tonify both spleen and kidney, and a preparation method thereof.

[0006] The technical solution adopted by the present invention to solve its technical problem is:

[0007] In one aspect of the present invention, a traditional Chinese medicine composition for preventing and treating sarcopenia is provided, comprising the following traditional Chinese medicine raw materials in parts by weight: 1-30 parts of ginseng, 1-30 parts of Panax notoginseng, 1-12 parts of yam, 1-15 parts of Poria, 0.2-9 parts of cinnamon bark or 1-9 parts of epimedium, 1-9 parts of red dates, and 0.5-3 parts of licorice.

[0008] The preferred ratio of the above raw materials is: 2-20 parts of ginseng, 1.5-20 parts of Panax notoginseng, 2-8 parts of yam, 2-10 parts of Poria, 0.5-6 parts of cinnamon or 1.5-7 parts of epimedium, 2-6 parts of red dates, and 1-2 parts of licorice. The most preferred ratio of the above raw materials is: 2 parts of ginseng, 1.5 parts of Panax notoginseng, 4 parts of Chinese yam, 3 parts of Yun Poria, 0.5 parts of cinnamon or 1.5 parts of epimedium, 3 parts of large red dates, and 1 part of raw licorice.

[0009] Other medicines can also be added to the above-mentioned traditional Chinese medicines to form a variety of prescriptions to exert different degrees of efficacy.

[0010] As a preferred technical solution of the present invention, the raw materials include one or more of the following medicines: 1-20 parts of Codonopsis pilosula and 1-40 parts of dried tangerine peel, in addition to 1-30 parts of ginseng, 1-30 parts of Panax notoginseng, 1-12 parts of yam, 1-15 parts of Poria, 0.2-9 parts of cinnamon bark or 1-9 parts of epimedium, 1-9 parts of red dates and 0.5-3 parts of licorice.

[0011] The preferred ratio of the above raw materials is: 2 parts of ginseng, 1.5 parts of Panax notoginseng, 4 parts of Chinese yam, 3 parts of Yun Fuling, 0.5 parts of cinnamon or 1.5 parts of epimedium, 3 parts of red dates, 1 part of raw licorice, 2 parts of Codonopsis, and 4 parts of dried tangerine peel.

[0012] The Chinese medicine used in the present invention has definite curative effect, is safe and reliable, and has significant curative effect. Among them, ginseng is the root of ginseng of the Araliaceae family, and the root contains a variety of ginsenosides. Ginseng is warm in nature, sweet, slightly bitter and slightly warm in taste. It enters the spleen and lung meridians. Panax notoginseng is the dried root of Panax notoginseng (Burk.) FHChen of the Araliaceae family, which has the effects of removing blood stasis and stopping bleeding, promoting blood circulation and relieving pain. Panax notoginseng is produced in Jilong, Zhangmu, Dingjie, Medog and Linzhi. It grows in broad-leaved forests or mixed coniferous gardens at an altitude of 2300-3200 (-3300) meters. It is also found in Nepal, Bhutan and northeastern India. It is another name for Panax notoginseng, which is equivalent to Panax notoginseng. Yam is the dried rhizome of Dioscorea zingiberensis, a plant of the Dioscoreaceae family. Huai yam is native to Pingyao and Jiexiu, Shanxi Province, and is now distributed in North China, Northwest China and Jiangxi, Hunan and other regions in the Yangtze River Basin. Those mainly produced in Xinxiang, Henan Province are preferred, and are called Huai yam. Poria cocos is the dried sclerotium of the fungus Poria cocos (Schw.) Wolf of the Polyporaceae family. Yunnan Poria cocos refers to the authentic medicinal material of Poria cocos produced in Yunnan. Official cinnamon comes from the trunk of cassia bark, has a slender shape, and has high medicinal value. Epimedium is the dried leaves of the plants Epimedium, Epimedium sagittatum, Epimedium pubescens, or Epimedium koreana of the Berberidaceae family. Red dates are the fruits of the jujube tree, which are warm in nature and sweet in taste. They belong to the spleen, stomach, and heart meridians. They have the effects of tonifying the middle and replenishing qi, nourishing blood and calming the mind. They are suitable for weak spleen and stomach, loss of appetite, abdominal distension and pain, nausea and vomiting, diarrhea, etc. Licorice is the dried root and rhizome of Glycyrrhizauralensis Fisch., Glycyrrhiza inflata Bat. or Glycyrrhiza glabra L. of the Leguminosae family. It is sweet and flat in nature, and enters the heart, lung, spleen and stomach meridians. It has the effects of replenishing qi and tonifying the middle, relieving acute pain, alleviating medicinal properties, removing phlegm and relieving cough, and detoxifying. Raw licorice belongs to a kind of licorice, and licorice belongs to a large category. In traditional Chinese medicine, licorice is divided into raw licorice and roasted licorice. Raw licorice is cool in nature and tends to clear heat. Its main effects are clearing heat and detoxifying, moistening the lungs and removing phlegm. Codonopsis is a qi-tonifying medicine, which is the dried root of Codonopsis pilosula, Codonopsis pilosula or Codonopsis pilosula of the Campanulaceae family. Tangerine peel is a qi-regulating medicine, which is the dried and mature peel of the citrus plant and its cultivated varieties of the Rutaceae family.

[0013] This prescription targets the characteristics of spleen and kidney deficiency in TCM's "atrophy syndrome" of elderly sarcopenia, and adopts the method of spleen and kidney tonification and qi and blood regulation. Specifically: based on the TCM theory of "monarch, minister, assistant and envoy", it focuses on tonifying qi, benefiting the kidney and strengthening the spleen, while taking into account the effects of activating blood circulation and harmonizing. Through multi-level compatibility, it achieves a dynamic balance of "tonifying without stagnation, warming without dryness", and its pharmacological action is reflected in the coordinated regulation of multiple targets. Red ginseng, as the main drug, has the effects of greatly replenishing vital energy, consolidating the body and promoting the production of body fluid; Panax notoginseng, as the assistant drug, plays a synergistic role, assisting red ginseng in enhancing the effect of promoting blood circulation and removing blood stasis; cinnamon bark and epimedium, as assistant drugs, are alternatives to each other. If it is kidney yang deficiency syndrome, cinnamon bark is selected, and if it is kidney essence deficiency syndrome, epimedium is selected; Chinese yam, as the adjuvant, is used to strengthen the spleen and consolidate the foundation, and prevent the tonic drugs from causing greasiness; Yun Fuling, as the adjuvant, is used to promote diuresis and eliminate dampness to balance the warm and dry nature of cinnamon bark or epimedium; red dates, as the guiding drug, have a synergistic effect, and can replenish blood and prevent greasiness; raw licorice, as the guiding drug, has a harmonizing effect, and in this prescription it has the function of alleviating the harsh nature of other drugs.

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

[0015] In another aspect of the present invention, a method for preparing the Chinese medicine composition is provided, comprising the following steps: weighing the raw medicine according to the weight portion to prepare a tea.

[0016] In another aspect of the present invention, a method for preparing the Chinese medicine composition is provided, comprising the following steps: weighing the raw material medicine according to the weight portion, adding 6-12 times of water for decoction, slightly boiling for 30min-90min, filtering, adding 4-10 times of water to the residue, slightly boiling for 30min-60min, filtering; combining the two decoctions, concentrating, adding appropriate amounts of auxiliary materials to the extract or the dried extract powder to obtain a granular preparation of the Chinese medicine composition.

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

[0018] The beneficial effect of the present invention is that according to the physiological and pathological characteristics of sarcopenia, the present invention provides a Chinese medicine formula with significant efficacy and no toxic side effects for the pathological basis of paralysis caused by spleen and kidney deficiency and qi and blood imbalance, reflecting the characteristics of traditional Chinese medicine syndrome differentiation and treatment of sarcopenia.

[0019] The concept, specific implementation methods and technical effects of the present invention will be further described below in conjunction with the accompanying drawings to fully understand the purpose, characteristics and effects of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a graph showing the results of a cell activity experiment conducted in Example 15 to detect the effect of the Chinese herbal compound of the present invention on the activity of mouse myoblasts C2C12. DETAILED DESCRIPTION

[0021] The present invention is described below in conjunction with specific embodiments. The technical methods involved therein, unless otherwise specified in the present invention, are all technical methods conventionally used by those skilled in the art, and thus will not be described in detail.

[0022] Example 1 Preparation of the pharmaceutical preparation of the present invention

[0023] 2 grams of ginseng, 1.5 grams of Panax notoginseng, 4 grams of Chinese yam, 3 grams of Yun Fuling, 0.5 grams of official cinnamon, 3 grams of large red dates, and 1 gram of raw licorice.

[0024] Weigh the medicinal materials according to the above prescription, add 10 times water to decoct, slightly boil for 60 minutes, filter, add 8 times water to the residue, slightly boil for 60 minutes, filter. Combine the two decoctions, decompress and concentrate to a relative density of 1.25-1.30, use an appropriate amount of dextrin as the base material, spray the concentrate, granulate in a fluidized bed in one step, make 1000g, package, 10g per package, twice a day, 2 packages each time.

[0025] Example 2 Preparation of the pharmaceutical preparation of the present invention

[0026] 2 grams of ginseng, 1.5 grams of Panax notoginseng, 4 grams of Chinese yam, 3 grams of Yun Fuling, 1.5 grams of Epimedium, 3 grams of red dates, and 1 gram of raw licorice.

[0027] Weigh the medicinal materials according to the above prescription, add 10 times water to decoct, slightly boil for 60 minutes, filter, add 8 times water to the residue, slightly boil for 60 minutes, filter. Combine the two decoctions, concentrate under reduced pressure, dry, add appropriate amount of lactose, dextrin, magnesium stearate to the extract powder, mix evenly, granulate, make 1000g, package, 10g per package, 3 times a day, 2 packages each time.

[0028] Example 3 Preparation of the pharmaceutical preparation of the present invention

[0029] 1 gram of ginseng, 1 gram of Panax notoginseng, 1 gram of yam, 1 gram of Poria cocos, 0.2 gram of cinnamon bark, 1 gram of red dates, and 0.5 gram of licorice.

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

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

[0032] 1 gram of ginseng, 1 gram of Panax notoginseng, 1 gram of yam, 1 gram of Poria cocos, 1 gram of epimedium, 1 gram of red dates, and 0.5 gram of licorice.

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

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

[0035] 30 grams of ginseng, 30 grams of Panax notoginseng, 12 grams of yam, 15 grams of Poria cocos, 9 grams of cinnamon bark, 9 grams of red dates, and 3 grams of licorice.

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

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

[0038] 30 grams of ginseng, 30 grams of Panax notoginseng, 12 grams of yam, 15 grams of Poria cocos, 9 grams of epimedium, 9 grams of red dates, and 3 grams of licorice.

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

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

[0041] 2 grams of ginseng, 1.5 grams of Panax notoginseng, 2 grams of yam, 2 grams of Poria cocos, 0.5 grams of cinnamon bark, 2 portions of red dates, and 1 gram of licorice.

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

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

[0044] 2 grams of ginseng, 1.5 grams of Panax notoginseng, 2 grams of yam, 2 grams of Poria cocos, 1.5 grams of epimedium, 2 portions of red dates, and 1 gram of licorice.

[0045] The above raw materials are made into tea.

[0046] Example 9 Preparation of the pharmaceutical preparation of the present invention (water decoction)

[0047] 20 grams of ginseng, 20 grams of Panax notoginseng, 8 grams of yam, 10 grams of Poria cocos, 6 grams of cinnamon bark, 6 grams of red dates, and 2 grams of licorice.

[0048] The above raw materials were decocted and extracted twice with water. For the first decoction, 1500 ml of water was added for 60 minutes, and 150 ml of the remaining juice was taken. For the second decoction, 1000 ml of water was added for 45 minutes. For the third decoction, 1000 ml of water was added for 45 minutes, and 150 ml of the remaining juice was taken. Orally take twice a day, 150 ml each time.

[0049] Example 10 Preparation of the pharmaceutical preparation of the present invention (water decoction)

[0050] 20 grams of ginseng, 20 grams of Panax notoginseng, 8 grams of yam, 10 grams of Poria cocos, 7 grams of Epimedium, 6 grams of red dates, and 2 grams of licorice.

[0051] The above raw materials were decocted and extracted twice with water. For the first decoction, 1500 ml of water was added for 60 minutes, and 150 ml of the remaining juice was taken. For the second decoction, 1000 ml of water was added for 45 minutes. For the third decoction, 1000 ml of water was added for 45 minutes, and 150 ml of the remaining juice was taken. Orally take twice a day, 150 ml each time.

[0052] Example 11 Preparation of the pharmaceutical preparation of the present invention (tablets)

[0053] 2g ginseng, 1.5g notoginseng, 4g Chinese yam, 3g Yun Fuling, 0.5g cinnamon, 3g red dates, 1g raw licorice, 2g Codonopsis, 4g tangerine peel.

[0054] Weigh the medicinal materials according to the above prescription, add 10 times water to decoct, slightly boil for 60 minutes, filter, add 10 times water to the residue, slightly boil for 30 minutes, filter. Combine the two decoctions, concentrate under reduced pressure, dry, add appropriate amount of dextrin and magnesium stearate to the extract powder, mix evenly, granulate, make 1000g, dry at 80℃ and press into tablets to obtain tablets, each tablet is 0.5 grams, take orally 3 times a day, 5 tablets each time.

[0055] Example 12 Preparation of the pharmaceutical preparation of the present invention (tablets)

[0056] 2g ginseng, 1.5g notoginseng, 4g Chinese yam, 3g Yun Fuling, 1.5g epimedium, 3g red dates, 1g raw licorice, 2g Codonopsis pilosula, 4g tangerine peel.

[0057] Weigh the medicinal materials according to the above prescription, add 10 times water to decoct, slightly boil for 60 minutes, filter, add 10 times water to the residue, slightly boil for 30 minutes, filter. Combine the two decoctions, concentrate under reduced pressure, dry, add appropriate amount of dextrin and magnesium stearate to the extract powder, mix evenly, granulate, make 1000g, dry at 80℃ and press into tablets to obtain tablets, each tablet is 0.5 grams, take orally 3 times a day, 5 tablets each time.

[0058] Example 13 Preparation of the pharmaceutical preparation of the present invention (capsules)

[0059] 1g of ginseng, 1g of Panax notoginseng, 1g of Chinese yam, 1g of Yun Fuling, 0.5g of cinnamon bark, 1g of red dates, 0.5g of raw licorice, 2g of Codonopsis pilosula, and 4g of tangerine peel.

[0060] Weigh the medicinal materials according to the above prescription, add 10 times water to decoct, slightly boil for 60 minutes, filter, add 10 times water to the residue, slightly boil for 30 minutes, filter. Combine the two decoctions, concentrate under reduced pressure, dry, add appropriate amount of dextrin and magnesium stearate to the extract powder, mix evenly, granulate, make 1000g, add appropriate amount of micro powder silica gel, mix evenly, put into capsules, 0.4g per capsule, take orally 3 times a day, 5 capsules each time.

[0061] Example 14 Preparation of the pharmaceutical preparation of the present invention (capsules)

[0062] 1g of ginseng, 1g of Panax notoginseng, 1g of Chinese yam, 1g of Yun Fuling, 1.5g of Epimedium, 1g of red dates, 0.5g of raw licorice, 2g of Codonopsis pilosula, and 4g of dried tangerine peel.

[0063] Weigh the medicinal materials according to the above prescription, add 10 times water to decoct, slightly boil for 60 minutes, filter, add 10 times water to the residue, slightly boil for 30 minutes, filter. Combine the two decoctions, concentrate under reduced pressure, dry, add appropriate amount of dextrin and magnesium stearate to the extract powder, mix evenly, granulate, make 1000g, add appropriate amount of micro powder silica gel, mix evenly, put into capsules, 0.4g per capsule, take orally 3 times a day, 5 capsules each time.

[0064] Embodiment 15

[0065] The beneficial effects of the present invention are further described below through pharmacological studies:

[0066] In order to detect the effect of the Chinese medicinal compound of the present invention on the activity of mouse myoblast C2C12, the following cell activity experiment was performed.

[0067] 1. Materials and methods:

[0068] 1.1 Experimental Materials

[0069] 1.1.1 C2C12 cells were purchased from the Shanghai Cell Bank, Chinese Academy of Sciences.

[0070] 1.1.2 Main reagents and instruments

[0071] 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 luminescent cell viability detection kit (C0057M, Beyotime). Spark multifunctional microplate reader (Tecan).

[0072] 1.2 Experimental methods

[0073] 1.2.1 Viability measurement of serum starved cell model

[0074] Step 1, seeding C2C12 myoblasts on a cell culture dish and culturing them in proliferation medium (DMEM + 10% fetal bovine serum, 1% penicillin / streptomycin [P / S]), changing the medium at least every 2 days;

[0075] Step 2: C2C12 cells (0.5×10 5 cells / well) were seeded on a 96-well cell culture dish and cultured overnight in proliferation medium;

[0076] Step 3, the C2C12 cells were divided into a normal group, a starvation group and an experimental group, wherein the C2C12 cells in the normal group were cultured in a proliferation medium, the C2C12 cells in the starvation group were cultured in a serum-free medium (DMEM, 1% penicillin / streptomycin [P / S]), and the C2C12 cells in the experimental group were cultured in a serum-free medium, and the extract of the compound prepared in Example 1 (hereinafter referred to as Compound 1) and the extract of the compound prepared in Example 2 (hereinafter referred to as Compound 2) were added to the culture medium at different concentrations (2.5, 5, 10, 25, 50, 75, 100 μg / mL) and cultured for 48 hours.

[0077] Step 4: Add 100 μl CellTiter-Lumi TM Plus II luminescence detection reagent, the chemiluminescence value (RLU) of each well was detected by a multifunctional microplate reader. Each experiment was repeated 3 times, with 6 replicate wells each time.

[0078] 1.2.2 Viability measurement of dexamethasone cell injury model

[0079] Step 1, seeding C2C12 myoblasts on a cell culture dish and culturing them in proliferation medium (DMEM + 10% fetal bovine serum, 1% penicillin / streptomycin [P / S]), changing the medium at least every 2 days;

[0080] Step 2: C2C12 cells (0.125×10 5 cells / well) were seeded on a 96-well cell culture dish and cultured in proliferation medium for 24 h;

[0081] Step 3, the C2C12 cells were divided into a normal group, a dexamethasone group and an experimental group, wherein 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 (2.5, 5, 10, 25, 50, 75, 100 μg / mL) of drug extracts of compound 1 and compound 2 were added to the culture medium, respectively, and cultured for 48 hours.

[0082] Step 4: Add 100 μl CellTiter-Lumi TM Plus II luminescence detection reagent, the chemiluminescence value of each well was detected by a multifunctional microplate reader. Each experiment was repeated 3 times, with 6 replicate wells each time.

[0083] 2. Statistics and Analysis

[0084] The results are as follows Figure 1As 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 cell injury model.

[0085] like Figure 1 As shown, both compound 1 and compound 2 have certain protective activity on serum-starved and dexamethasone-injured C2C12 cells, among which compound 1 with a concentration higher than 10 μg / mL has a significant effect on improving the vitality of serum-starved and dexamethasone-injured C2C12 cells; compound 2 with a concentration higher than 25 μg / mL has a significant effect on improving the vitality of serum-starved C2C12 cells, and compound 2 with a concentration higher than 10 μg / mL has a significant effect on improving the vitality of dexamethasone-injured C2C12 cells.

[0086] Therefore, the compound ratios of Example 1 and Example 2 can significantly improve the survival rate of C2C12 cells under serum starvation and dexamethasone injury, indicating that the drug of this patent has a good effect in improving sarcopenia.

[0087] Example 16

[0088] In order to investigate the effect of this prescription drug on sarcopenia model mice, the following experiment was conducted.

[0089] 1. Materials

[0090] 1.1 Experimental animals

[0091] Male KM mice, 6-8 weeks old, weighing 26-30 g, were purchased from Shanghai Model Organisms Technology Co., Ltd.

[0092] Mice were group-housed in cages of 5 each. The experiment began after 7 days of adaptive feeding in an environment with a room temperature of 20-25°C and a humidity of 30-60%. During the feeding period, feed and water were supplied without distinction. During the feeding process, the diet, activity, coat color, defecation, and mental state of the mice were observed at any time. The body weight was measured at the same time every day. All animals were kept in accordance with the "Regulations on the Management of Experimental Animals" formulated by the State Science and Technology Commission.

[0093] 1.2 Experimental drugs

[0094] Dexamethasone sodium phosphate injection, purchased from Shanghai Pharmaceutical Group Co., Ltd., was used as the model group.

[0095] The prescription drug is prepared using the following compound ratio, specifically:

[0096] Compound 1 (same prescription as Example 1): 2g ginseng, 1.5g Panax notoginseng, 4g Chinese yam, 3g Yun Fuling, 0.5g cinnamon bark, 3g red dates, 1g raw liquorice;

[0097] Compound 2 (same prescription as Example 2): 2g ginseng, 1.5g Panax notoginseng, 4g Chinese yam, 3g Yun Fuling, 1.5g Epimedium, 3g red dates, 1g raw liquorice;

[0098] Compound 3 (same prescription as Example 12): 2g ginseng, 1.5g ginseng notoginseng, 4g Chinese yam, 3g Yun Fuling, 1.5g epimedium, 3g red dates, 1g raw liquorice, 2g codonopsis, 4g tangerine peel;

[0099] Compound 4 (same prescription as Example 11): 2g ginseng, 1.5g Panax notoginseng, 4g Chinese yam, 3g Yun Fuling, 0.5g cinnamon bark, 3g red dates, 1g raw liquorice, 2g Codonopsis pilosula, 4g dried orange peel;

[0100] Compound 5 (same prescription as Example 3): 1g ginseng, 1g notoginseng, 1g yam, 1g poria, 0.2g cinnamon bark, 1g red dates, 0.5g liquorice;

[0101] Compound 6 (same prescription as Example 6): 30g ginseng, 30g Panax notoginseng, 12g yam, 15g Poria, 9g epimedium, 9g red dates, and 3g liquorice.

[0102] 1.3 Experimental instruments

[0103] Dual-energy X-ray absorptiometry (iNSiGHT VET DXA, OsteoSys, KOREAN); YLS-13A muscle strength tester, Jinan Yiyan Technology Co., Ltd.

[0104] 2. Methods

[0105] 2.1 Modeling

[0106] After adaptive feeding, the male KM mice were randomly divided into groups, weighed and numbered. The negative control group (control, blank control) 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 distinction, and the bedding was changed twice a week to keep the living environment clean and hygienic.

[0107] 2.2 Grouping and Dosing

[0108] After 2 weeks of dexamethasone sodium phosphate injection, 56 male KM mice were randomly divided into model group, compound group 1 (4 g / kg), compound group 2 (4 g / kg), compound group 3 (4 g / kg), compound group 4 (4 g / kg), compound group 5 (4 g / kg), and compound group 6 (4 g / kg), with 8 mice in each group; 8 KM mice of the same age served as the negative control group.

[0109] Each compound extract was dissolved in 0.3% sodium carboxymethyl cellulose solution. Each administration group (including: compound 1 group (4g / kg), compound 2 group (4g / kg), compound 3 group (4g / kg), compound 4 group (4g / kg), compound 5 group (4g / kg), compound 6 group (4g / kg)) was given 4g / kg by gavage for 6 consecutive weeks, once a day, with a gavage volume of 0.2mL / 10g. The negative control group and the model group were given 0.3% sodium methyl cellulose solution by gavage for 6 consecutive weeks, once a day, with a gavage volume of 0.2mL / 10g.

[0110] 2.3 Detection indicators and methods

[0111] 2.3.1 Mouse muscle strength measurement

[0112] The forelimb grip strength of mice was measured using a YLS-13A 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 back by their tails, with their posture parallel to the surface of the table, until they released the mesh. The peak force exerted by the mouse limbs was recorded in grams (g). Evaluations were performed every 0, 2, 4, and 6 weeks, and each mouse was tested three times, and the average was taken and weighted for statistical analysis.

[0113] 2.3.2 Body composition of mice

[0114] Three days before the mice were killed, dual-energy X-ray absorptiometry (DEXA) was used to measure body composition, including body fat content, muscle content, bone mineral content, etc. The data were analyzed using the software provided by the manufacturer.

[0115] 2.3.3 Statistical methods

[0116] All data were expressed as mean ± SD, and the data were subjected to one-way ANOVA, and multiple comparisons were performed using Tukey post-hoc test. In the results, P < 0.05 indicated that the two groups of data had significant differences, P < 0.01 indicated that the two groups of data had very significant differences, and P < 0.001 indicated that the two groups of data had extremely significant differences. Graphpad Prism 10.1.2 was used as the statistical software.

[0117] 3. Results

[0118] 3.1 General observation of experimental animals

[0119] The negative control group was in good general condition, with smooth and shiny fur and no obvious irritability. Eating and drinking were normal, defecation was normal, weight increased steadily, and reaction was quick. Before the injection of dexamethasone, the mice in each group were in good condition, with shiny fur and agile activities. After the injection of dexamethasone began, the subcutaneous injection model group gradually showed irritability, slow reaction, listlessness, fluffy and dull fur, and a surge in weight. Some mice had necrosis at the tip of their tails and fell off automatically.

[0120] 3.2 Effects of each compound on the muscle strength of model mice

[0121] The results are shown in Table 1. Before administration, compared with the control group, the muscle strength of each group was significantly decreased (P < 0.001); there was no significant difference in the muscle strength of the model mice among the groups, indicating that the experimental system is reliable and the groups are comparable.

[0122] After 2 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 muscle strength of each compound group showed an upward trend, and there was no significant difference compared with the model group;

[0123] After 4 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 muscle strength of compound group 1 and compound group 2 increased most significantly, with a statistical difference of P < 0.001; compared with the model group, the muscle strength of compound group 5 and compound group 6 increased significantly, with a statistical difference of P < 0.01; compared with the model group, the muscle strength of compound group 3 and compound group 4 also increased significantly, with a statistical difference of P < 0.05.

[0124] After 6 weeks of administration, compared with the control group, the muscle strength of the model group was significantly decreased (P < 0.001); compared with the model group, the muscle strength of compound group 1, compound group 2, compound group 4, compound group 5, and compound group 6 increased most significantly, with a statistical difference of P < 0.001, among which the mean muscle strength of compound group 1 was the highest, which was 6.60 ± 0.40; compared with the model group, the muscle strength of compound group 3 increased more significantly, with a statistical difference of P < 0.01.

[0125] It can be seen that compound group 1, compound group 2, compound group 3, compound group 4, compound group 5, and compound group 6 all have obvious effects on improving the muscle strength of sarcopenia model mice, among which compound group 1 has the best effect.

[0126] Table 1 Effect of this prescription on muscle strength of model mice (X±SD)

[0127] Before administration 2 weeks after administration 4 weeks after administration 6 weeks after administration Control group 5.92±0.34 5.98±0.38 6.11±0.35 6.79±0.46 Model Group <![CDATA[3.63±0.31 *** ]]> <![CDATA[3.76±0.53 *** ]]> <![CDATA[3.84±0.30 *** ]]> <![CDATA[4.25±0.35 *** <!-- 8 -->]]> Compound 1 group <![CDATA[3.49±0.36 *** ]]> 4.68±0.35 <![CDATA[5.32±0.22 ### ]]> <![CDATA[6.60±0.40 ### ]]> Compound 2 group <![CDATA[3.65±0.56 *** ]]> 4.54±0.54 <![CDATA[5.02±0.45 ### ]]> <![CDATA[6.52±0.70 ### ]]> Compound 3 groups <![CDATA[3.42±0.57 *** ]]> 4.46±0.63 <![CDATA[4.72±0.20 # ]]> <![CDATA[5.68±0.64 ## ]]> Compound 4 groups <![CDATA[3.22±0.44 *** ]]> 4.45±0.40 <![CDATA[4.79±0.57 # ]]> <![CDATA[5.91±0.51 ### ]]> Compound 5 groups <![CDATA[3.27±0.26 *** ]]> 4.01±0.33 <![CDATA[4.80±0.54 ## ]]> <![CDATA[6.37±0.42 ### ]]> Compound 6 groups <![CDATA[3.71±0.55 *** ]]> 4.20±0.47 <![CDATA[4.93±0.29 ## ]]> <![CDATA[6.38±0.70 ### ]]>

[0128] Note: Compared with the control group, the model group ***P<0.001; compared with the model group, the drug-treated group #P<0.05, ##P<0.01, ###P<0.001.

[0129] 3.3 Effects of this prescription drug on body fat content and muscle content in sarcopenia model mice

[0130] The results after 6 weeks of administration are shown in Table 6. Compared with the control group, the body fat content of the model group was significantly increased (P < 0.001); compared with the model group, the body fat content of compound group 1 and compound group 2 decreased most significantly, with a statistical difference of P < 0.001, among which the body fat content of compound group 1 was the lowest, at 6.07% ± 2.49%; compared with the model group, the body fat content of compound group 3, compound group 4, and compound group 5 decreased more significantly, with a statistical difference of P < 0.01; compared with the model group, the decrease in body fat content of compound group 6 was also statistically significant (P < 0.05).

[0131] Compared with the control group, the muscle content of the model group was significantly decreased (P < 0.001); compared with the model group, the muscle content of compound 1 group increased most significantly, with a statistical difference of P < 0.001; compared with the model group, the muscle content of compound 2 group also increased significantly, with a statistical difference of P < 0.01; the muscle content of compound 3 group, compound 4 group, compound 5 group, and compound 6 group showed an upward trend, with no significant difference compared with the model group.

[0132] It can be seen that Compound Group 1, Compound Group 2, Compound Group 3, Compound Group 4, Compound Group 5, and Compound Group 6 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.

[0133] Table 2 Effects of each compound group of drugs on body fat content and muscle content in sarcopenia model mice (X±SD)

[0134] Body fat content (100%) Muscle content (%) Control group 4.46±1.76 91.15±2.98 Model Group <![CDATA[12.90±2.87 *** ]]> <![CDATA[82.96±2.28 *** ]]> Compound 1 group <![CDATA[6.07±2.49 ### ]]> <![CDATA[90.59±2.76 ### ]]> Compound 2 group <![CDATA[6.16±1.76 ### ]]> <![CDATA[89.73±2.13 ## ]]> Compound 3 groups <![CDATA[7.19±2.67 ## ]]> 82.65±2.25 Compound 4 groups <![CDATA[6.84±2.11 ## ]]> 85.54±2.54 Compound 5 groups <![CDATA[6.98±2.01 ## ]]> 81.33±2.27 Compound 6 groups <![CDATA[7.70±1.82 # ]]> 86.12±2.03

[0135] Note: Compared with the control group, the model group *P<0.05, **P<0.01; compared with the model group, the drug-treated group #P<0.05, ##P<0.01, ###P<0.001.

[0136] In summary, by verifying that the compound of each embodiment has obvious improvements on the muscle strength, body fat content and muscle content of sarcopenia mice, it is proved that the Chinese medicine composition of the present invention does have a better technical effect of preventing and treating sarcopenia.

[0137] The preferred specific embodiments of the present invention are described in detail above. It should be understood that ordinary technicians in the field can make many modifications and changes based on the concept of the present invention without creative work. Therefore, all technical solutions that can be obtained by technicians in the technical field based on the concept of the present invention through logical analysis, reasoning or limited experiments on the basis of the prior art should be within the scope of protection determined by the claims.

Claims

1. A Chinese medicine composition for preventing and treating sarcopenia, characterized in that: The medicine is mainly prepared from the following raw materials in parts by weight: 1-30 parts of ginseng, 1-30 parts of notoginseng, 1-12 parts of yam, 1-15 parts of tuckahoe, 0.2-9 parts of cinnamon bark or 1-9 parts of epimedium, 1-9 parts of red dates and 0.5-3 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: 2-20 parts of ginseng, 1.5-20 parts of notoginseng, 2-8 parts of yam, 2-10 parts of tuckahoe, 0.5-6 parts of cinnamon bark or 1.5-7 parts of epimedium, 2-6 parts of red dates and 1-2 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: 2 parts of ginseng, 1.5 parts of notoginseng, 4 parts of Chinese yam, 3 parts of Yunfuling, 0.5 parts of cinnamon bark or 1.5 parts of epimedium, 3 parts of red dates and 1 part of raw licorice.

4. The Chinese medicine composition according to claim 1, characterized in that: The raw materials also include the following medicines: 1-20 parts of Codonopsis pilosula and 1-40 parts of dried tangerine peel.

5. The Chinese medicine composition according to claim 4, characterized in that: The medicine is mainly prepared from the following raw materials in parts by weight: 2 parts of ginseng, 1.5 parts of notoginseng, 4 parts of Chinese yam, 3 parts of Yunfuling, 0.5 parts of cinnamon bark or 1.5 parts of epimedium, 3 parts of red dates, 1 part of raw licorice, 2 parts of codonopsis pilosula, and 4 parts of tangerine peel.

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 to prepare a tea.

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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