Traditional Chinese medicine compound for treating sarcopenia and preparation and application of fermentation liquor of traditional Chinese medicine compound

Abstract

The invention discloses a traditional Chinese medicine compound for treating sarcopenia and preparation and application of fermentation liquor of the traditional Chinese medicine compound, and belongs to the field of traditional Chinese medicine application. The traditional Chinese medicine compound for treating sarcopenia is prepared from the following raw materials in parts by weight: 10 to 30 parts of radix astragali seu hedysari, 5 to 25 parts of radix codonopsis, 5 to 25 parts of herba cistanche, 5 to 25 parts of folium cortex eucommiae, 5 to 25 parts of rhizoma polygonati, 5 to 25 parts of radix angelicae sinensis, 5 to 25 parts of fructus crataegi and 5 to 25 parts of roasted fructus hordei germinatus. The traditional Chinese medicine compound fermentation liquor for treating sarcopenia is prepared by the following steps: uniformly mixing the traditional Chinese medicines in parts by weight, adding water, decocting to obtain decoction, and concentrating the decoction to obtain concentrated liquor; inoculating lactobacillus johnsonii, lactobacillus fermentum and bifidobacterium adolescentis into the concentrated solution, and fermenting to obtain fermentation liquor; the traditional Chinese medicine composition has the advantages that the pathological state of sarcopenia is improved through the traditional Chinese medicine composition, and meanwhile a new scheme is provided for the probiotic fermented traditional Chinese medicine decoction.

Description

Technical Field

[0001] This application relates to the field of traditional Chinese medicine applications, and in particular to a traditional Chinese medicine compound for treating sarcopenia and the preparation and application of its fermentation broth. Background Technology

[0002] Sarcopenia is a syndrome characterized by reduced skeletal muscle mass, decreased muscle strength, and impaired function. It is commonly seen in individuals undergoing 5-fluorouracil-induced chemotherapy for cancer, those with chronic alcohol consumption, the elderly, and those with chronic diseases. Sarcopenia not only leads to decreased motor function and quality of life but also increases the risk of infection, falls, and death. However, clinical treatment faces many limitations: nutritional supplementation can only slow down muscle loss and cannot reverse atrophy; adherence to exercise rehabilitation is extremely low in patients who are weak after chemotherapy or have liver damage; although chemotherapy drugs can provide short-term relief of local symptoms, they have hepatotoxicity and nephrotoxicity and target only a single point, making it difficult to cover the multiple pathological links of gut microbiota imbalance, muscle inflammation, and metabolic disorders, especially failing to simultaneously improve 5-fluorouracil-induced muscle damage and systemic metabolic imbalance.

[0003] Traditional Chinese medicine (TCM) classifies sarcopenia under the categories of "asthenia" and "atrophy," with its core pathogenesis being spleen and kidney deficiency, insufficient qi and blood, and malnourishment of tendons and bones. Clinical manifestations include reduced skeletal muscle mass, decreased muscle strength, and impaired motor function, often accompanied by systemic symptoms such as fatigue and poor appetite. Treatment focuses on strengthening the spleen and kidneys, nourishing blood and strengthening muscles, emphasizing the nourishment and repair of muscles through regulating organ function. Currently, TCM interventions for sarcopenia in clinical practice often employ traditional decoction-based compound herbal formulas. While these formulas are formulated based on the principles of syndrome differentiation and treatment, they suffer from a delayed onset of efficacy.

[0004] Probiotic fermentation technology provides a key pathway for optimizing traditional Chinese medicine (TCM) compound formulas. Its core value lies in transforming TCM components through the metabolic activities of probiotics: on the one hand, it can decompose macromolecular components in TCM, improving the dissolution rate of effective ingredients; on the other hand, it can generate new active substances, improving the absorption efficiency and bioavailability of TCM compound formulas. However, most TCM compound formulas are not currently optimized using probiotic fermentation technology, and the application of probiotics is mostly limited to single strains, lacking effective integration with TCM compound formulas. In particular, for 5-fluorouracil-induced sarcopenia, there is a lack of research on improving TCM compound formulas based on probiotic fermentation technology, making it difficult to address the shortcomings of traditional treatments through technological integration. Summary of the Invention

[0005] In order to obtain a traditional Chinese medicine compound and its fermentation liquid that can be used to treat sarcopenia, this application provides a method for preparing and applying a traditional Chinese medicine compound and its fermentation liquid for treating sarcopenia.

[0006] In the first aspect, this application provides a traditional Chinese medicine compound for treating sarcopenia, employing the following technical solution: A traditional Chinese medicine compound for treating sarcopenia contains the following ingredients in parts by weight: Astragalus membranaceus 10-30 parts, Codonopsis pilosula 5-25 parts, Cistanche deserticola 5-25 parts, Eucommia ulmoides leaf 5-25 parts, Polygonatum sibiricum 5-25 parts, Angelica sinensis 5-25 parts, Crataegus pinnatifida 5-25 parts, and roasted malt 5-25 parts.

[0007] The pharmacological effects of each component in the above-mentioned traditional Chinese medicine compound: Astragalus: It has a sweet taste and slightly warm nature, and enters the lung and spleen meridians. It has the functions of tonifying qi and raising yang, consolidating the exterior and stopping sweating, promoting diuresis and reducing swelling, generating fluids and nourishing blood, and promoting the elimination of toxins and pus. It is used for qi deficiency and fatigue, poor appetite and loose stools, sinking of middle qi, chronic diarrhea and rectal prolapse, hematochezia and metrorrhagia, spontaneous sweating due to exterior deficiency, edema due to qi deficiency, internal heat and thirst, blood deficiency and chlorosis, hemiplegia, numbness and pain, carbuncles and boils that are difficult to ulcerate, and chronic ulcers that do not heal.

[0008] Codonopsis pilosula: sweet in taste, neutral in nature, enters the lung and spleen meridians; it has the effects of tonifying the middle energizer and replenishing qi, strengthening the spleen and benefiting the lungs, generating fluids and nourishing blood. It is used for spleen and lung qi deficiency, poor appetite and fatigue, cough and wheezing, qi and blood deficiency, sallow complexion, palpitations and shortness of breath, thirst due to fluid depletion, and internal heat and thirst.

[0009] Cistanche deserticola: It is sweet and salty in taste, warm in nature, and enters the kidney and large intestine meridians. It has the effects of tonifying kidney yang, nourishing essence and blood, and moistening the intestines to relieve constipation. It is used for kidney yang deficiency, deficiency of essence and blood, impotence and infertility, soreness and weakness of the waist and knees, weakness of muscles and bones, and constipation due to intestinal dryness.

[0010] Eucommia ulmoides leaves: sweet in taste, warm in nature, and enter the liver and kidney meridians; they have the effects of tonifying the liver and kidneys and strengthening tendons and bones, and are used for lower back and knee pain, weakness of tendons and bones, dizziness and vertigo caused by liver and kidney deficiency.

[0011] Polygonatum: Sweet, neutral. It enters the spleen, lung, and kidney meridians; it has the effects of tonifying qi and nourishing yin, strengthening the spleen, moistening the lungs, and benefiting the kidneys. It is used for spleen and stomach qi deficiency, fatigue, stomach yin deficiency, dry mouth and poor appetite, lung deficiency and dry cough, consumptive cough with hemoptysis, deficiency of essence and blood, soreness and weakness of the lower back and knees, premature graying of hair, and internal heat and thirst.

[0012] Angelica sinensis: sweet and pungent in taste, warm in nature, and enters the liver, heart, and spleen meridians; it has the effects of nourishing blood and promoting blood circulation, regulating menstruation and relieving pain, and moistening the intestines and relieving constipation. It is used for blood deficiency and chlorosis, dizziness and palpitations, irregular menstruation, amenorrhea and dysmenorrhea, abdominal pain due to deficiency and cold, rheumatic arthralgia, traumatic injuries, carbuncles and sores, and constipation due to intestinal dryness.

[0013] Hawthorn: It tastes sour and sweet, and is slightly warm in nature. It enters the spleen, stomach, and liver meridians. It has the effects of promoting digestion and strengthening the stomach, promoting blood circulation and removing blood stasis, and astringing and stopping diarrhea. It is used for food stagnation, abdominal distension, diarrhea and abdominal pain, amenorrhea due to blood stasis, postpartum blood stasis, stabbing pain in the heart and abdomen, hernia pain, and hyperlipidemia.

[0014] Roasted malt: It has a sweet taste and neutral properties, and enters the spleen, stomach, and liver meridians. It has the effects of promoting qi circulation and digestion, strengthening the spleen and stomach, reducing lactation and relieving bloating. It is used for indigestion, abdominal distension and pain, spleen deficiency and poor appetite, milk stasis, breast distension and pain, weaning in women, liver stagnation and hypochondriac pain, and liver and stomach qi pain.

[0015] By adopting the above technical solution, this combination of eight Chinese herbs follows the principle of principal, assistant, adjuvant, and guide herbs, yielding significant benefits: The principal herbs, Astragalus and Codonopsis, invigorate the spleen and replenish qi to strengthen the acquired constitution, laying the material foundation for muscle growth; the assistant herbs, Cistanche, Eucommia, and Polygonatum, nourish the kidneys and strengthen muscles to replenish the innate essence, contributing to strong bones and muscles; the adjuvant herb, Angelica, nourishes and invigorates blood, clearing the meridians to promote the nourishment of muscles by qi and blood; and the guiding herbs, Crataegus pinnatifida and roasted malt, aid digestion and harmonize the stomach, helping the absorption of the effective components of the principal, assistant, and adjuvant herbs while also assisting in the generation of qi and blood. The combined effects of these herbs are to invigorate the spleen and kidneys, nourish blood, and strengthen muscles. All are food-grade medicinal materials, balancing therapeutic efficacy with long-term safety, and can improve muscle malnutrition caused by spleen and kidney deficiency and insufficient qi and blood from multiple dimensions.

[0016] Optionally, the traditional Chinese medicine compound contains the following ingredients in parts by weight: 20-30 parts Astragalus membranaceus, 10-20 parts Codonopsis pilosula, 10-20 parts Cistanche deserticola, 10-20 parts Eucommia ulmoides leaf, 10-20 parts Polygonatum sibiricum, 5-15 parts Angelica sinensis, 10-20 parts Crataegus pinnatifida, and 10-20 parts roasted malt.

[0017] Optionally, the traditional Chinese medicine compound contains the following ingredients in parts by weight: 15 parts Astragalus membranaceus, 10 parts Codonopsis pilosula, 10 parts Cistanche deserticola, 10 parts Eucommia ulmoides leaf, 10 parts Polygonatum sibiricum, 5 parts Angelica sinensis, 10 parts Crataegus pinnatifida, and 10 parts roasted malt.

[0018] Secondly, this application provides the use of a traditional Chinese medicine compound for treating sarcopenia in the preparation of drugs for treating or preventing sarcopenia.

[0019] Thirdly, this application provides a traditional Chinese medicine compound preparation for treating sarcopenia, employing the following technical solution: A traditional Chinese medicine compound preparation for treating sarcopenia, prepared from a concentrated liquid, wherein the preparation comprises one or more of decoctions, pills, and tablets; the concentrated liquid is prepared through the following steps: Astragalus membranaceus, Codonopsis pilosula, Cistanche deserticola, Eucommia ulmoides leaves, Polygonatum sibiricum, Angelica sinensis, hawthorn and roasted malt were mixed and decocted with water to obtain a decoction. The decoction was then concentrated to obtain a concentrated liquid.

[0020] Decoctions include concentrated liquids, granules, and oral liquids; pills include honey pills, paste pills, water pills, drop pills, and wax pills; tablets are solid preparations made by uniformly mixing concentrated liquids and excipients and then pressing them together.

[0021] By adopting the above technical solution, the decoction process allows the components of each Chinese herbal medicine to be continuously released and dissolved, thereby obtaining the effective components in the Chinese herbal medicine compound.

[0022] Optionally, weigh out the following ingredients according to the above weight fractions: Astragalus membranaceus, Codonopsis pilosula, Cistanche deserticola, Eucommia ulmoides leaves, Polygonatum sibiricum, Angelica sinensis, Crataegus pinnatifida, and roasted malt. Mix them well and decoct with water at least twice. When decocting twice, decoct for 30-60 minutes for the first time and 30-60 minutes for the second time. Filter to obtain the decoction. Combine the decoctions and concentrate them to a relative density of 1.1-1.3 g / mL to obtain the concentrated solution.

[0023] Fourthly, this application provides a traditional Chinese medicine compound fermentation liquid for treating sarcopenia, employing the following technical solution: A traditional Chinese medicine compound fermentation liquid for treating sarcopenia is prepared through the following steps: Mix Astragalus membranaceus, Codonopsis pilosula, Cistanche deserticola, Eucommia ulmoides leaves, Polygonatum sibiricum, Angelica sinensis, Crataegus pinnatifida and roasted malt in proportion to weight, add water and decoct to obtain decoction, and then concentrate the decoction to obtain concentrated liquid. Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis were inoculated into a concentrated solution for fermentation to obtain a fermentation broth.

[0024] By adopting the above technical solution, based on the concentrated Chinese medicine liquid, synergistic fermentation of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis is carried out. Through the synergistic effect of the strains, the utilization rate and bioactivity of the effective components of Chinese medicine are improved, which can significantly improve sarcopenia. Specific manifestations include inhibiting weight loss, constipation symptoms and colonic mucosal damage, prolonging the weight-bearing swimming time to improve muscle endurance, improving skeletal muscle atrophy, reducing the content of inflammatory cytokines in muscle tissue and increasing liver glycogen content. At the same time, it improves the systemic metabolism and damage serum markers of sarcopenia mice, including lactate, lactate dehydrogenase, creatine kinase, creatinine, alanine aminotransferase (ALT), and aspartate aminotransferase (AST).

[0025] Optionally, the preparation methods of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis include the following steps: (1) Lactobacillus johnsonii strain, Lactobacillus fermentum strain and Bifidobacterium adolescentis strain were inoculated into the first culture medium and activated to obtain Lactobacillus johnsonii activated bacterial solution, Lactobacillus fermentum activated bacterial solution and Bifidobacterium adolescentis activated bacterial solution, respectively. (2) The activated bacterial solutions of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis were inoculated into the second culture medium and cultured to obtain Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis respectively. (3) Take a portion of the bacterial suspension from the Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis suspension respectively, dilute them in a gradient with buffer, and inoculate them on the third culture medium and culture them until single colonies are visible. (4) After centrifuging the Lactobacillus johnsonii culture, Lactobacillus fermentum culture and Bifidobacterium adolescentis culture obtained from (3), collect the bacterial precipitate to obtain Lactobacillus johnsonii cells, Lactobacillus fermentum cells and Bifidobacterium adolescentis cells.

[0026] Optionally, the inoculation concentration of *Lactobacillus johnsonii* is 2 × 10⁻⁶. 8 -3×10 10 CFU / mL, the inoculation concentrations of *Lactobacillus fermentum* and *Bifidobacterium adolescentis* were 1×10⁻⁶ CFU / mL, respectively. 8 -1.5×10 10 CFU / mL.

[0027] By adopting the above technical solution, controlling the proportion of the above three strains, and supplementing with concentrated Chinese medicine for synergistic fermentation, the fermentation effect can be effectively promoted, thereby improving sarcopenia.

[0028] Optionally, the inoculation amount of the Lactobacillus johnsonii strain is 1-3% of the volume of the concentrate.

[0029] Optionally, the inoculation concentration ratio of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis is (1-2):(1-2):(1-2).

[0030] Optionally, the inoculation concentration ratio of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis is 2:1:1.

[0031] Fifthly, this application provides a traditional Chinese medicine compound fermented liquid preparation for treating sarcopenia, employing the following technical solution: This application provides a traditional Chinese medicine compound preparation for treating sarcopenia, using the following technical solution: A traditional Chinese medicine compound fermented liquid preparation for treating sarcopenia, obtained from fermented liquid, the preparation comprising one or more of decoctions, pills and tablets.

[0032] In summary, this application has the following beneficial effects: 1. This application uses eight traditional Chinese medicines—Astragalus membranaceus, Codonopsis pilosula, Cistanche deserticola, Eucommia ulmoides leaf, Polygonatum sibiricum, Angelica sinensis, Crataegus pinnatifida, and roasted malt—in accordance with the principles of principal, assistant, adjuvant, and guiding herbs, resulting in significant benefits: The principal herbs, Astragalus membranaceus and Codonopsis pilosula, invigorate the spleen and replenish qi to strengthen the acquired constitution, laying a material foundation for muscle growth; the assistant herbs, Cistanche deserticola, Eucommia ulmoides leaf, and Polygonatum sibiricum, nourish the kidneys and strengthen tendons to replenish innate essence, contributing to strong bones and muscles; the adjuvant herb, Angelica sinensis, nourishes and invigorates blood, clears the meridians, and promotes the nourishment of muscles by qi and blood; the guiding herbs, Crataegus pinnatifida and roasted malt, aid digestion and harmonize the stomach, helping the absorption of the effective components of the principal, assistant, and adjuvant herbs, and assisting in the generation of qi and blood. The combined effects of these herbs are to invigorate the spleen and kidneys, nourish blood, and strengthen muscles. All are food-grade medicinal materials, balancing therapeutic efficacy with long-term safety, and can improve muscle malnutrition caused by spleen and kidney deficiency and insufficient qi and blood from multiple dimensions.

[0033] 2. Based on concentrated traditional Chinese medicine extract, synergistic fermentation of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis is carried out. Through the synergistic effect of strains, the utilization rate and bioactivity of the effective components of traditional Chinese medicine are improved, which can significantly improve sarcopenia. Specific manifestations include inhibiting weight loss, constipation symptoms and colonic mucosal damage, prolonging the weight-bearing swimming time to improve muscle endurance, improving skeletal muscle atrophy, reducing the content of inflammatory cytokines in muscle tissue and increasing liver glycogen content. At the same time, it improves the systemic metabolism and serum markers of damage in sarcopenic mice, including lactate, lactate dehydrogenase, creatine kinase, creatinine, alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Attached Figure Description

[0034] Figure 1 Changes in mouse body weight during pharmacodynamic evaluation; Figure 2 The change in weight-bearing swimming time of mice during pharmacodynamic evaluation; Figure 3 For pharmacodynamic evaluation, comparisons were made between wet recombinant groups of mouse soleus muscle. Figure 4 For comparison of mouse plasma parameters between groups during pharmacodynamic evaluation; Figure 5 For pharmacodynamic evaluation, the comparison of liver glycogen levels between mouse groups was performed. Figure 6 For pharmacodynamic evaluation, HE staining of mouse colon pathological sections was performed between groups. Figure 7 This is a comparison between groups of HE-stained mouse soleus muscle tissue sections during pharmacodynamic evaluation.

[0035] Figure 8 This is a comparison of slow muscle protein fluorescence staining in mouse soleus muscle tissue sections during pharmacodynamic evaluation. Detailed Implementation

[0036] The following is in conjunction with the appendix Figure 1-8 This application will be described in further detail.

[0037] Lactobacillus johnsonii, accession number BNCC135265, source number CGMCC1.3255.

[0038] Lactobacillus fermentum, with accession number BNCC194390 and source number CICC22827.

[0039] Bifidobacterium adolescentis, with accession number BNCC185973 and source number CICC6070.

[0040] The sources of the above-mentioned microbial species are used to support the scheme of this application to provide sufficient disclosure, but are not limited to the sources listed.

[0041] Example 1.1 A traditional Chinese medicine compound for treating sarcopenia contains the following ingredients in parts by weight: Astragalus membranaceus 15 parts, Codonopsis pilosula 10 parts, Cistanche deserticola 10 parts, Eucommia ulmoides leaf 10 parts, Polygonatum sibiricum 10 parts, Angelica sinensis 5 parts, Crataegus pinnatifida 10 parts, and roasted malt 10 parts.

[0042] A method for preparing a traditional Chinese medicine compound for treating sarcopenia includes the following steps: Weigh out the following ingredients according to the above weight fractions: Astragalus membranaceus, Codonopsis pilosula, Cistanche deserticola, Eucommia ulmoides leaves, Polygonatum sibiricum, Angelica sinensis, Crataegus pinnatifida, and roasted malt. Mix them well and decoct twice with water. The first decoction is 45 minutes and the second decoction is 45 minutes. Filter to obtain the decoction liquid. Combine the two decoction liquids and concentrate them to a relative density of 1.2 g / mL to obtain preparation concentrate A.

[0043] Example 1.2 A traditional Chinese medicine compound for treating sarcopenia contains the following ingredients in parts by weight: Astragalus membranaceus 10 parts, Codonopsis pilosula 5 parts, Cistanche deserticola 5 parts, Eucommia ulmoides leaf 5 parts, Polygonatum sibiricum 5 parts, Angelica sinensis 5 parts, Crataegus pinnatifida 5 parts, and roasted malt 5 parts.

[0044] A method for preparing a traditional Chinese medicine compound for treating sarcopenia includes the following steps: Weigh out the following ingredients according to the above weight fractions: Astragalus membranaceus, Codonopsis pilosula, Cistanche deserticola, Eucommia ulmoides leaves, Polygonatum sibiricum, Angelica sinensis, Crataegus pinnatifida, and roasted malt. Mix them well and decoct twice with water. The first decoction is 30 minutes and the second decoction is 30 minutes. Filter to obtain the decoction liquid. Combine the two decoction liquids and concentrate them to a relative density of 1.1 g / mL to obtain preparation concentrate B.

[0045] Example 1.3 A traditional Chinese medicine compound for treating sarcopenia contains the following ingredients in parts by weight: Astragalus membranaceus 30 parts, Codonopsis pilosula 25 parts, Cistanche deserticola 25 parts, Eucommia ulmoides leaf 25 parts, Polygonatum sibiricum 25 parts, Angelica sinensis 25 parts, Crataegus pinnatifida 25 parts, and roasted malt 25 parts.

[0046] A method for preparing a traditional Chinese medicine compound for treating sarcopenia includes the following steps: Weigh out the following ingredients according to the above weight fractions: Astragalus membranaceus, Codonopsis pilosula, Cistanche deserticola, Eucommia ulmoides leaves, Polygonatum sibiricum, Angelica sinensis, Crataegus pinnatifida, and roasted malt. Mix them well and decoct twice with water. The first decoction is 60 minutes and the second decoction is 60 minutes. Filter to obtain the decoction liquid. Combine the two decoction liquids and concentrate them to a relative density of 1.3 g / mL to obtain the preparation concentrate C. Example

[0047] A traditional Chinese medicine compound fermentation broth is prepared through the following steps: (1) Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis were inoculated into the first culture medium and activated to obtain Lactobacillus johnsonii activated bacterial solution, Lactobacillus fermentum activated bacterial solution and Bifidobacterium adolescentis activated bacterial solution respectively.

[0048] The first culture medium includes MRS liquid medium and Bifidobacterium liquid medium. Lactobacillus johnsonii and Lactobacillus fermentum are inoculated into MRS liquid medium, and Bifidobacterium adolescentis is inoculated into Bifidobacterium liquid medium.

[0049] The activation time was 24 hours and the activation temperature was 37℃.

[0050] (2) The activated bacterial solutions of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis were inoculated into the second culture medium and cultured to obtain Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis, respectively.

[0051] The second culture medium includes MRS liquid medium and Bifidobacterium liquid medium. Lactobacillus johnsonii and Lactobacillus fermentum are inoculated into MRS liquid medium, while Bifidobacterium adolescentis is inoculated into Bifidobacterium liquid medium.

[0052] The inoculum size was 1%, the culture temperature was 37℃, and the culture time was 24 hours.

[0053] (3) Take a portion of the bacterial solution from Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis respectively, dilute them in a gradient with buffer, and inoculate them into the third culture medium. Cultivate until single colonies are visible, and calculate the bacterial concentration of Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis respectively.

[0054] The buffer used was PBS buffer, serially diluted to 1×10⁻⁶. -7 The third culture medium included MRS agar and Bifidobacterium agar. *Lactobacillus johnsonii* and *Lactobacillus fermentum* strains were inoculated onto MRS agar, while *Bifidobacterium adolescentis* strains were inoculated onto Bifidobacterium agar. The concentration of *Lactobacillus johnsonii* cultured to this level was 1 × 10⁻⁶. 9 The concentrations of CFU / mL for Lactobacillus fermentation culture and Bifidobacterium adolescentis culture were 5 × 10⁻⁶. 8 CFU / mL.

[0055] (4) The Lactobacillus johnsonii culture, Lactobacillus fermentum culture and Bifidobacterium adolescentis culture obtained from S3 culture were centrifuged and the bacterial precipitates were collected to obtain Lactobacillus johnsonii cells, Lactobacillus fermentum cells and Bifidobacterium adolescentis cells.

[0056] (5) Take Lactobacillus johnsonii cells, Lactobacillus fermentum cells, and Bifidobacterium adolescentis cells and inoculate them into the formulation concentrate A in Example 1.1. The concentration of Lactobacillus johnsonii cells is 1×10⁻⁶. 9 CFU / mL, the concentrations of Lactobacillus fermentum and Bifidobacterium adolescentis were 5×10⁻⁶. 8 The concentration ratio of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis was 2:1:1, measured in CFU / mL (colony forming units). The cultures were resuspended and combined in concentrate A, and the concentrations were adjusted. The culture temperature was 37℃, and the culture time was 24 hours to obtain the fermentation broth, which is denoted as fermentation broth A. Example

[0057] A traditional Chinese medicine compound fermentation broth is prepared through the following steps: (1) Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis were inoculated into the first culture medium and activated to obtain Lactobacillus johnsonii activated bacterial solution, Lactobacillus fermentum activated bacterial solution and Bifidobacterium adolescentis activated bacterial solution respectively.

[0058] The first culture medium includes MRS liquid medium and Bifidobacterium liquid medium. Lactobacillus johnsonii and Lactobacillus fermentum are inoculated into MRS liquid medium, and Bifidobacterium adolescentis is inoculated into Bifidobacterium liquid medium.

[0059] The activation time was 24 hours and the activation temperature was 37℃.

[0060] (2) The activated bacterial solutions of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis were inoculated into the second culture medium and cultured to obtain Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis, respectively.

[0061] The second culture medium includes MRS liquid medium and Bifidobacterium liquid medium. Lactobacillus johnsonii and Lactobacillus fermentum are inoculated into MRS liquid medium, while Bifidobacterium adolescentis is inoculated into Bifidobacterium liquid medium.

[0062] The inoculum size was 1%, the culture temperature was 37℃, and the culture time was 24 hours.

[0063] (3) Take a portion of the bacterial solution from Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis respectively, dilute them in a gradient with buffer, and inoculate them into the third culture medium. Cultivate until single colonies are visible, and calculate the bacterial concentration of Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis respectively.

[0064] The buffer used was PBS buffer, serially diluted to 1×10⁻⁶. -7The third culture medium included MRS agar and Bifidobacterium agar. *Lactobacillus johnsonii* and *Lactobacillus fermentum* strains were inoculated onto MRS agar, while *Bifidobacterium adolescentis* strains were inoculated onto Bifidobacterium agar. The concentration of *Lactobacillus johnsonii* cultured to this level was 1 × 10⁻⁶. 9 The concentrations of CFU / mL for Lactobacillus fermentation culture and Bifidobacterium adolescentis culture were 5 × 10⁻⁶. 8 CFU / mL.

[0065] (4) The Lactobacillus johnsonii culture, Lactobacillus fermentum culture and Bifidobacterium adolescentis culture obtained from S3 culture were centrifuged and the bacterial precipitates were collected to obtain Lactobacillus johnsonii cells, Lactobacillus fermentum cells and Bifidobacterium adolescentis cells.

[0066] (5) Inoculate Lactobacillus johnsonii cells, Lactobacillus fermentum cells, and Bifidobacterium adolescentis cells into the concentrate B of the preparation in Example 1.2, and culture until the Lactobacillus johnsonii cell concentration is 1×10⁻⁶. 9 The concentrations of CFU / mL for Lactobacillus fermentation culture and Bifidobacterium adolescentis culture were 5 × 10⁻⁶. 8 The concentration ratio of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis was 2:1:1, measured in CFU / mL (according to colony-forming units). The cultures were resuspended and combined in concentrate B, and the concentrations were adjusted. The culture temperature was 37℃, and the culture time was 24 hours to obtain the traditional Chinese medicine fermentation broth, which is denoted as fermentation broth B. Example

[0067] A traditional Chinese medicine compound fermentation broth is prepared through the following steps: (1) Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis were inoculated into the first culture medium and activated to obtain Lactobacillus johnsonii activated bacterial solution, Lactobacillus fermentum activated bacterial solution and Bifidobacterium adolescentis activated bacterial solution respectively.

[0068] The first culture medium includes MRS liquid medium and Bifidobacterium liquid medium. Lactobacillus johnsonii and Lactobacillus fermentum are inoculated into MRS liquid medium, and Bifidobacterium adolescentis is inoculated into Bifidobacterium liquid medium.

[0069] The activation time was 24 hours and the activation temperature was 37℃.

[0070] (2) The activated bacterial solutions of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis were inoculated into the second culture medium and cultured to obtain Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis, respectively.

[0071] The second culture medium includes MRS liquid medium and Bifidobacterium liquid medium. Lactobacillus johnsonii and Lactobacillus fermentum are inoculated into MRS liquid medium, while Bifidobacterium adolescentis is inoculated into Bifidobacterium liquid medium.

[0072] The inoculum size was 1%, the culture temperature was 37℃, and the culture time was 24 hours.

[0073] (3) Take a portion of the bacterial solution from Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis respectively, dilute them in a gradient with buffer, and inoculate them into the third culture medium. Cultivate until single colonies are visible, and calculate the bacterial concentration of Lactobacillus johnsonii, Lactobacillus fermentum and Bifidobacterium adolescentis respectively.

[0074] The buffer used was PBS buffer, serially diluted to 1×10⁻⁶. -7 The third culture medium included MRS agar and Bifidobacterium agar. *Lactobacillus johnsonii* and *Lactobacillus fermentum* strains were inoculated onto MRS agar, while *Bifidobacterium adolescentis* strains were inoculated onto Bifidobacterium agar. The concentration of *Lactobacillus johnsonii* cultured to this level was 1 × 10⁻⁶. 9 The concentrations of CFU / mL for Lactobacillus fermentation culture and Bifidobacterium adolescentis culture were 5 × 10⁻⁶. 8 CFU / mL.

[0075] (4) The Lactobacillus johnsonii culture, Lactobacillus fermentum culture and Bifidobacterium adolescentis culture obtained from S3 culture were centrifuged and the bacterial precipitates were collected to obtain Lactobacillus johnsonii cells, Lactobacillus fermentum cells and Bifidobacterium adolescentis cells.

[0076] (5) Inoculate Lactobacillus johnsonii cells, Lactobacillus fermentum cells, and Bifidobacterium adolescentis cells into the concentrate C of the preparation in Example 1.3, and culture until the Lactobacillus johnsonii cell concentration is 1×10⁻⁶. 9 The concentrations of CFU / mL for Lactobacillus fermentation culture and Bifidobacterium adolescentis culture were 5 × 10⁻⁶. 8 The concentration ratio of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis was 2:1:1, measured in CFU / mL (colony forming units). The cultures were resuspended and combined in concentrate C, and the concentrations were adjusted. The culture temperature was 37℃, and the culture time was 24 hours to obtain the fermentation broth of traditional Chinese medicine, which is denoted as fermentation broth C.

[0077] Comparative Example 1 The difference between this traditional Chinese medicine compound fermentation broth and Example 2 is that the fermented bacteria do not contain Lactobacillus johnsonii, but only Lactobacillus fermentum and Bifidobacterium adolescentis in a 1:1 concentration ratio. The resulting fermentation broth is denoted as Fermentation Broth D.

[0078] Comparative Example 2 The difference between this traditional Chinese medicine compound fermentation broth and Example 2 is that the fermented bacteria do not contain Lactobacillus fermentum, but only Lactobacillus johnsonii and Bifidobacterium adolescentis in a 1:1 concentration ratio. The resulting fermentation broth is denoted as Fermentation Broth E.

[0079] Comparative Example 3 The difference between this traditional Chinese medicine compound fermentation broth and Example 2 is that the fermented bacteria do not contain Bifidobacterium adolescentis, but only contain Lactobacillus johnsonii and Lactobacillus fermentum in a 1:1 concentration ratio. The resulting fermentation broth is denoted as Fermentation Broth F.

[0080] Application Example 1-1 A granule formulation comprising the following steps: Take the concentrated liquid A of the traditional Chinese medicine compound preparation in Example 1.1, add dextrin (the mass of dextrin is twice the mass of concentrated liquid A), add 95% ethanol as a wetting agent to make a soft material, granulate it through a 16-mesh sieve, dry it, and granulate it to make granules.

[0081] Application Example 1-2 An oral liquid formulation comprising the following steps: Take the concentrated liquid A of the traditional Chinese medicine compound preparation in Example 1.1, add purified water to adjust the relative density of concentrated liquid A to 1.05 g / mL, then add sucrose at 5% of the total weight of concentrated liquid A for flavoring, and add potassium sorbate at 0.01% of the total weight of concentrated liquid A as a preservative, and stir until completely dissolved; filter the above mixture through a 0.22 μm microporous membrane to remove impurities; take the filtered clear liquid, fill it into 10 mL sterile oral liquid bottles, sterilize by high pressure steam at 121℃ for 15-20 min, cool and seal to obtain the oral liquid preparation.

[0082] Application Example 2-1 A granule formulation comprising the following steps: Take the fermentation broth A of the traditional Chinese medicine compound from Example 2, add dextrin (the mass of dextrin is twice the mass of fermentation broth A), use 95% ethanol as a wetting agent to make a soft mass, granulate it through a 16-mesh sieve, dry it, and granulate it to make granules.

[0083] Application Example 2-2 An oral liquid comprising the following steps: Take the fermentation broth A of the traditional Chinese medicine compound from Example 2, add purified water to adjust the relative density of fermentation broth A to 1.05 g / mL, then add sucrose at 5% of the total weight of fermentation broth A for flavoring, and add potassium sorbate at 0.01% of the total weight of fermentation broth A as a preservative, and stir until completely dissolved; filter the above mixture through a 0.22 μm microporous membrane to remove impurities; take the filtered clear liquid, fill it into 10 mL sterile oral liquid bottles, sterilize by high-pressure steam at 121℃ for 15-20 min, cool and seal to obtain the oral liquid preparation.

[0084] Pharmacological trials The animal experiments were approved by the Animal Ethics Committee of Southern Medical University (Approval No. L202312007, Date: 2023-12-14). Male Balb / c mice (20 ± 2 g) were purchased from the Experimental Animal Center of Southern Medical University and acclimatized to the environment under standard laboratory conditions for 7 days before the experiment. Seventy-two male Balb / c mice were randomly divided into 9 groups and acclimatized for 7 days. After the acclimatization period, mice in each group were administered gavage from day 8 to day 15 (treatment period), and injected with 25 mg / kg 5-fluorouracil from day 8 to day 12 (modeling period). Muscle strength and function (dynamic behavior) of the mice were assessed at the beginning, middle and end of the experiment; after fasting for 12 h on day 16, the mice were euthanized by intraperitoneal injection of 1% sodium pentobarbital and then by cervical dislocation. Plasma, colon tissue, gastrocnemius muscle and soleus muscle of the mice were collected for the determination of various indicators. The specific grouping is as follows (Table 1): (1) Normal group: Throughout the experimental period, mice were provided with normal food and water. Each mouse was given 0.2 mL of physiological saline by gavage and 0.2 mL of physiological saline by intraperitoneal injection daily; (2) Model group: Throughout the experimental period, mice were provided with normal feed and water. During the modeling period, each mouse was given 0.2 mL of physiological saline by gavage and 25 mg / kg of 5-fluorouracil by intraperitoneal injection daily; (3) Concentrated Solution A Group: Throughout the experimental period, mice were provided with normal feed and water. During the modeling period, each mouse was given 0.2 mL of concentrated solution A by gavage and 25 mg / kg of 5-fluorouracil by intraperitoneal injection daily; (4) Fermentation broth group A: Throughout the experimental period, mice were provided with normal feed and water. During the modeling period, each mouse was given 0.2 mL of fermentation broth A by gavage and 25 mg / kg of 5-fluorouracil by intraperitoneal injection daily; (5) Fermentation broth group B: Throughout the experimental period, mice were provided with normal feed and water. During the modeling period, each mouse was given 0.2 mL of fermentation broth B by gavage and 25 mg / kg of 5-fluorouracil by intraperitoneal injection daily; (6) Fermentation broth group C: Throughout the experimental period, mice were provided with normal feed and water. During the modeling period, each mouse was given 0.2 mL of fermentation broth C by gavage and 25 mg / kg of 5-fluorouracil by intraperitoneal injection daily; (7) Fermentation broth group D: Throughout the experimental period, mice were provided with normal feed and water. During the modeling period, each mouse was given 0.2 mL of fermentation broth D by gavage and 25 mg / kg of 5-fluorouracil by intraperitoneal injection daily; (8) Fermentation broth group E: Throughout the experimental period, mice were provided with normal feed and water. During the modeling period, each mouse was given 0.2 mL of fermentation broth E by gavage and 25 mg / kg of 5-fluorouracil by intraperitoneal injection daily; (9) Fermentation broth group F: Throughout the experimental period, mice were provided with normal feed and water. During the modeling period, each mouse was given 0.2 mL of fermentation broth F by gavage and 25 mg / kg of 5-fluorouracil by intraperitoneal injection.

[0085] Table 1. Experimental Groups and Drug Solution Control Table Group Bacteria / Prescription Composition Source of medicine normal group — physiological saline Model group — physiological saline Concentrate Group A Example 1.1 Prescription Example 1 Fermentation broth group A Example 1.1 Prescription + Lactobacillus johnsonii : Lactobacillus fermentum : Bifidobacterium adolescentis = 2 : 1 : 1 Example 2 Fermentation broth group B Example 1.2 Prescription + Lactobacillus johnsonii : Lactobacillus fermentum : Bifidobacterium adolescentis = 2 : 1 : 1 Example 3 Fermentation broth group C Example 1.3 prescription + Lactobacillus johnsonii : Lactobacillus fermentum : Bifidobacterium adolescentis = 2 : 1 : 1 Example 4 Fermentation broth group D Example 1.1 Prescription + Lactobacillus fermentum : Bifidobacterium adolescentis = 1 : 1 Comparative Example 1 Fermentation broth group E Example 1.1 Prescription + Lactobacillus johnsonii : Bifidobacterium adolescentis = 1 : 1 Comparative Example 2 Fermentation broth group F Example 1.1 Prescription + Lactobacillus johnsonii : Lactobacillus fermentum = 1 : 1 Comparative Example 3 Table 2. Orthogonal experimental design table for combined probiotics Lactobacillus johnsonii Lactobacillus fermentation Bifidobacterium adolescentis 1 <![CDATA[5×10 8 CFU / mL]]> <![CDATA[5×10 8 CFU / mL]]> <![CDATA[5×10 8 CFU / mL]]> 2 <![CDATA[10 9 CFU / mL]]> <![CDATA[10 9 CFU / mL]]> <![CDATA[10 9 CFU / mL]]> Table 3. Results of orthogonal experiments on combined probiotics based on the formulation of Example 1.1

[0086] Pharmacodynamic evaluation This pharmacodynamic evaluation was conducted based on a 5-fluorouracil-induced sarcopenic Balb / c mouse model, according to the drug source corresponding to the experimental groups. The evaluation focused on weight recovery, muscle endurance, muscle mass protection, serum metabolic markers, and histopathological verification of the intervention effect.

[0087] like Figure 1 As shown, during the modeling period, 5-fluorouracil induced a decrease in mouse body weight, with significant differences among the treatment groups. The normal group showed a stable increase in body weight, while the model group experienced a continuous decrease. Example 1.1, as an unfermented traditional Chinese medicine compound preparation, achieved a stable inhibition of further weight loss by combining eight traditional Chinese medicines: Astragalus membranaceus, Codonopsis pilosula, Cistanche deserticola, Eucommia ulmoides leaf, Polygonatum sibiricum, Angelica sinensis, Crataegus pinnatifida, and roasted malt, demonstrating clear intervention value compared to the model group.

[0088] Example 2, relying on a synergistic fermentation system of *Lactobacillus johnsonii*, *Lactobacillus fermentum*, and *Bifidobacterium adolescentis*, inhibited weight loss during the modeling period and allowed for rapid recovery to near-normal levels in the later stages, demonstrating a more significant synergistic effect. While Comparative Examples 1-3, due to the absence of certain strains in their fermentation systems, could not achieve synergistic effects, they still inhibited weight loss by utilizing the efficacy of the traditional Chinese medicine compound itself and the partial effects of the remaining strains. Their effects were not significantly different from Example 1.1, failing to demonstrate the significant advantage of Example 2. The superior weight recovery effect of Example 2 compared to Comparative Examples 1-3 confirms that the synergistic fermentation of the three strains at the optimal ratio is more stable and efficient. The superior weight recovery effect of Example 2 compared to Examples 3 and 4 indicates that the dosage of the drug combination prescription in Example 2 is optimal.

[0089] like Figure 2As shown, swimming time was measured on day 0, day 5, and the last day. There were no differences among the groups before modeling; during the mid-modeling period, swimming time decreased in all groups, with the model group showing the largest decrease. Example 2 showed the smallest decrease due to the early onset of action of the active substances from the three-strain fermentation. Comparative Examples 1-3 showed similar decreases to Example 1. At the experimental endpoint, the normal group showed a continuous increase, while the model group continued to decrease. Example 2 showed the longest swimming time, approaching that of the normal group, with stable mouse stamina, demonstrating a significant benefit from synergistic fermentation. Although Example 1.1 was not as good as Example 2, it showed a significant improvement over the model group, with a significant reduction in the number of swimming pauses, proving that it can effectively improve muscle endurance without fermentation and possesses independent potential therapeutic value. Comparative Examples 1-3 also slightly improved swimming time, but the effect was not significantly different from Example 1.1, failing to achieve a breakthrough in synergistic effect, and mice still experienced a few pauses. Example 2 was superior to Comparative Examples 1-3, highlighting the synergistic effect of optimal ratio of three-strain fermentation. Example 2 was superior to Examples 3 and 4, demonstrating that the dosage of the drug combination prescription in Example 2 was appropriate.

[0090] like Figure 3 As shown, the soleus muscle weight test revealed that the normal group had sufficient muscle mass, while the model group showed significant atrophy. Example 2 utilized the synergistic fermentation of three bacteria to efficiently decompose the traditional Chinese medicine components, generating more active substances beneficial to muscle fiber synthesis. The muscle weight was closest to that of the normal group, essentially alleviating atrophy and demonstrating a significant synergistic effect. Example 1.1, relying on a complete traditional Chinese medicine formula, effectively increased muscle mass, showing significant improvement compared to the model group. Although the lack of fermentation resulted in a lower dissolution rate of the kidney-tonifying and muscle-strengthening components compared to Example 2, it still possessed a clear muscle mass protection effect and is the independent and effective basic solution in the patent. Comparative Examples 1-3, due to incomplete fermentation systems, only slightly increased muscle mass, with no significant difference in effect from Example 1.1, and could not achieve the significant recovery of muscle mass as seen in Example 2. Therefore, the synergistic fermentation of the traditional Chinese medicine components by the three bacteria is indispensable. Based on this, an orthogonal experiment was conducted (Table 3). The results showed that the optimal fermentation ratio for improving skeletal muscle mass was obtained when Lactobacillus johnsonii: Lactobacillus fermentum: Bifidobacterium adolescentis = 2:1:1. Furthermore, through a comparison of fermentation with three different dosages, the muscle mass of Example 2 was higher than that of Examples 3 and 4, confirming that it was the optimal dosage.

[0091] like Figure 4 and Figure 5As shown, serum metabolism and damage marker detection revealed that creatinine and lactate levels decreased in the model group, while lactate dehydrogenase, creatine kinase, alanine aminotransferase, and aspartate aminotransferase levels increased. Example 2, through the synergistic fermentation of three bacteria, enhanced the bioavailability of the traditional Chinese medicine components, regulated the metabolic network, significantly improved various indicators, repaired muscle membranes, and reduced liver damage, demonstrating a significant synergistic effect. Although Example 1.1 could not achieve the same level of regulation as Example 2, it effectively regulated some key indicators, showing a clear advantage over the model group, proving its ability to improve metabolic disorders and tissue damage, and is an independent scheme with potential therapeutic value. Comparative Examples 1-3 only slightly regulated the indicators, with no significant difference in effect from Example 1.1, failing to achieve synergistic effects, further confirming the unique value of the synergistic fermentation of three bacteria. In liver glycogen detection, Example 2 increased liver glycogen to near-normal levels, providing energy for muscles, followed by Example 1.1. The improvement effects of Comparative Examples 1-3 were similar to those of Example 1.1, none of which reached the level of Example 2, further highlighting the prominent role of bacterial strains in synergistic fermentation of traditional Chinese medicine prescriptions.

[0092] like Figure 6 As shown, colon pathological sections revealed that the normal group had intact mucosa, regular glands, and sufficient goblet cells; the model group showed significant mucosal erosion, glandular disorder, and inflammatory infiltration; Example 2, with its three-strain fermentation producing abundant active substances, exhibited the best repair effect, with the mucosa remaining largely intact and inflammation minimal; although Example 1.1 did not repair as well as Example 2, it still reduced mucosal erosion and inflammatory infiltration, and the number of goblet cells significantly increased compared to the model group, demonstrating its ability to independently improve the intestinal barrier and providing a foundation for treatment; Comparative Examples 1-3, due to insufficient fermented active substances, showed no significant difference in repair effect compared to Example 1.1, with mild mucosal erosion and inflammation remaining, failing to achieve the highly efficient repair of Example 2.

[0093] like Figure 7 As shown in the pathological sections of the soleus muscle, the normal group had regular muscle fibers, narrow interstices, and no inflammation; the model group had atrophied muscle fibers, wide interstices, breaks, and more inflammation; Example 2, with its three-strain fermentation, reduced inflammatory factors, and the muscle fibers were close to normal with no inflammation, showing a significant synergistic effect; Example 1, although still showing a small amount of inflammation and slight widening of interstices, was able to reduce the degree of muscle fiber atrophy, showing a significant repair advantage compared to the model group, confirming its independent protective effect on muscle fibers; Comparative Examples 1-3, due to the lack of fermentation anti-inflammatory products, did not show a significant difference in repair effect compared to Example 1.1, and the muscle fibers still showed slight atrophy, unable to repair breaks, thus failing to achieve synergistic effect.

[0094] like Figure 8As shown, immunofluorescence staining of slow-twitch muscle protein in soleus muscle revealed high and continuous expression of histone in normal groups; low and fragmented expression in the model group; in Example 2, three-strain fermentation activated the MyHC I expression pathway, resulting in near-normal protein expression and continuous signal, demonstrating a significant synergistic effect; although fragmentation still occurred locally in Example 1.1, the expression level of slow-twitch muscle protein was significantly higher than that in the model group, proving that it can promote slow-twitch muscle fiber repair without fermentation and has potential value for independent application; in Comparative Examples 1-3, due to insufficient fermentation active substances, the increase in protein expression was limited, and the effect was not significantly different from that of Example 1.1, with the fragmentation problem still being quite obvious, failing to achieve a breakthrough in synergistic effect.

[0095] In summary, Example 2, through the optimal dosage of the formula and the optimized three-strain synergistic fermentation scheme, maximized the spleen-tonifying, kidney-nourishing, blood-nourishing, and muscle-strengthening effects of traditional Chinese medicine through a complete fermentation system, achieving the best intervention effect. Its significant synergistic effect stemmed from the synergistic effect of the three probiotics. In contrast, Comparative Examples 1-3, due to the absence of any strain, lost the synergistic effect, and although effective, they could not achieve synergistic effects, failing to differentiate themselves from Example 1.1. Example 1.1, a basic scheme of unfermented traditional Chinese medicine compound, not only served as the core substrate for the fermentation optimization in Example 2 but also possessed independent and stable intervention value. It demonstrated clear advantages in dimensions such as weight protection, muscle endurance, muscle mass improvement, and tissue repair, possessing potential therapeutic value and providing an option for people who are not suitable for fermented preparations or prefer traditional Chinese medicine.

[0096] It is worth noting that the application of probiotic synergistic fermentation technology in this application provides a new path for the optimization of traditional Chinese medicine (TCM) compound prescriptions—enhancing the utilization rate and bioactivity of effective components of TCM through the synergistic effect of strains. This technology is not only applicable to the TCM compound prescription of this invention, but can also be extended to other TCM compound prescriptions with conditioning effects, providing a reference for the modernization and improvement of TCM and having broad application prospects. Examples 1 and 2 together constitute a diversified intervention system for sarcopenia based on the basic compound prescription and fermentation optimization. At the same time, the successful application of fermentation technology also provides a useful reference for technological innovation in the field of TCM.

[0097] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this specific embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A traditional Chinese medicine compound for treating sarcopenia, characterized in that, The raw materials include the following components in parts by weight: Radix Astragali 10-30 parts, Codonopsis 5-25 parts, Herba Cistanche 5-25 parts, Eucommia leaf 5-25 parts, Rhizoma Polygonati 5-25 parts, Angelica 5-25 parts, and Fructus Crataegi 5-25 parts.

2. The traditional Chinese medicine compound for treating sarcopenia according to claim 1, characterized in that: The raw materials include the following components in parts by weight: Radix Astragali 20-30 parts, Codonopsis 10-20 parts, Herba Cistanche 10-20 parts, Eucommia leaf 10-20 parts, Rhizoma Polygonati 10-20 parts, Angelica 5-15 parts, Fructus Crataegi 10-20 parts, and Fried Fructus Hordei 10-20 parts.

3. The traditional Chinese medicine compound for treating sarcopenia according to claim 1, characterized in that: The raw materials include the following components in parts by weight: Radix Astragali 15 parts, Codonopsis 10 parts, Herba Cistanche 10 parts, Eucommia leaf 10 parts, Rhizoma Polygonati 10 parts, Angelica 5 parts, Fructus Crataegi 10 parts, and Fried Fructus Hordei 10 parts.

4. Use of the Chinese medicine compound for treating sarcopenia according to any one of claims 1-3 in the preparation of a medicine for treating or preventing sarcopenia.

5. The Chinese medicine compound preparation for treating sarcopenia according to any one of claims 1-3, characterized in that, The preparation is prepared from a concentrated solution, and the preparation includes one or more of the following: a decoction, a pill, and a tablet. The raw materials are mixed and boiled in water to obtain a decoction, and the decoction is concentrated to obtain the concentrated solution.

6. A traditional Chinese medicine compound fermentation liquor for treating sarcopenia, characterized in that: The preparation is prepared from a concentrated solution, and the preparation includes one or more of the following: a decoction, a pill, and a tablet. The raw materials of the Chinese medicine compound for treating sarcopenia according to any one of claims 1-3 are mixed in parts by weight, boiled in water to obtain a decoction, and the decoction is concentrated to obtain the concentrated solution. Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis are inoculated into the concentrated solution to obtain a fermentation solution.

7. The traditional Chinese medicine compound fermentation liquor for treating sarcopenia according to claim 6, characterized in that: The preparation method of Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis includes the following steps: (1) Lactobacillus johnsonii, Lactobacillus fermentum, and Bifidobacterium adolescentis are inoculated into a first culture medium, activated, and Lactobacillus johnsonii activated bacteria solution, Lactobacillus fermentum activated bacteria solution, and Bifidobacterium adolescentis activated bacteria solution are obtained, respectively; (2) The Lactobacillus johnsonii activated bacteria solution, Lactobacillus fermentum activated bacteria solution, and Bifidobacterium adolescentis activated bacteria solution are inoculated into a second culture medium, cultured, and Lactobacillus johnsonii bacteria solution, Lactobacillus fermentum bacteria solution, and Bifidobacterium adolescentis bacteria solution are obtained, respectively; (3) Part of the Lactobacillus johnsonii bacteria solution, Lactobacillus fermentum bacteria solution, and Bifidobacterium adolescentis bacteria solution is gradient-diluted by a buffer solution and inoculated into a third culture medium, and cultured until single colonies are visible; (4) The Lactobacillus johnsonii bacteria solution, Lactobacillus fermentum bacteria solution, and Bifidobacterium adolescentis bacteria solution obtained by culture in (3) are centrifuged to collect bacterial body precipitates, and Lactobacillus johnsonii bacteria bodies, Lactobacillus fermentum bacteria bodies, and Bifidobacterium adolescentis bacteria bodies are obtained.

8. The traditional Chinese medicine compound fermentation liquor for treating sarcopenia according to claim 6, characterized in that: The inoculation concentration of *Lactobacillus johnsonii* was 2 × 10⁻� 8 -3×10 10 CFU / mL, the inoculation concentrations of *Lactobacillus fermentum* and *Bifidobacterium adolescentis* were 1×10⁻⁶ CFU / mL, respectively. 8 -1.5×10 10 CFU / mL. 9.The traditional Chinese medicine compound fermentation broth for treating sarcopenia according to claim 6, characterized in that: The inoculation concentration ratio of the Lactobacillus johnsonii bacteria bodies, Lactobacillus fermentum bacteria bodies, and Bifidobacterium adolescentis bacteria bodies is (1-2):(1-2):(1-2).

10. The Chinese medicinal compound fermentation broth preparation for treating sarcopenia according to claim 6, characterized in that, The preparation is prepared from a fermentation solution, and the preparation includes one or more of the following: a decoction, a pill, and a tablet.

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