Traditional Chinese medicine composition for strengthening spleen and building muscle, medicinal preparation and preparation method thereof

Abstract

The invention provides a traditional Chinese medicine composition for strengthening spleen and building muscle, a medicinal preparation and a preparation method thereof, and belongs to the technical field of medicines. The traditional Chinese medicine composition comprises the following components: Chinese yam, bighead atractylodes rhizome, sun-dried ginseng, hyacinth bean and malt. The traditional Chinese medicine composition and the pharmaceutical preparation provided by the invention have the effects of tonifying spleen and qi, promoting digestion and resolving dampness, and regulating middle warmer and increasing muscle, and can be used for treating senile sarcopenia and improving symptoms such as weakness of spleen and stomach, reduction of muscle strength, deficiency of qi and blood and the like.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a traditional Chinese medicine composition for invigorating the spleen and increasing muscle, a pharmaceutical preparation and a preparation method thereof. BACKGROUND

[0002] Sarcopenia, also known as muscle loss, is a syndrome caused by age-related decrease in muscle mass, strength or function. Skeletal muscle is the power of the human movement system, and muscle aging and atrophy are important signs of human aging, which can easily cause problems such as bone fracture and joint injury. Elderly people with muscle loss have difficulty standing, walk slowly and are prone to falling and fracturing. Sarcopenia also affects organ function and can cause heart and lung failure, and even death.

[0003] The occurrence of sarcopenia is related to factors such as hormone changes, malnutrition, insufficient exercise and chronic diseases, and needs to be intervened comprehensively through nutritional intervention, scientific exercise and chronic disease management. Different drugs are used to improve sarcopenia in traditional Chinese medicine. For example, Shan'you pills contain yam, atractylodes and ginseng; the efficacy is focused on tonifying qi and invigorating the spleen; the applicable symptoms are for weak spleen and stomach, less food and loose stools, fatigue and weakness; the medicine is pure and has no side effects, and is focused on strengthening the body and cultivating the essence. Jianpi pills contain atractylodes, ginseng, poria, hawthorn, god's recipe, malt, costus, tangerine peel, cardamom, goldthread, licorice, yam and nutmeg; the efficacy is focused on invigorating the spleen and stomach, and relieving food retention and diarrhea; the applicable symptoms are for spleen deficiency and food accumulation (abdominal distension, loss of appetite); the medicine tonifies the spleen while eliminating food retention and stagnation.

[0004] Due to the different symptoms of sarcopenia and the different focuses of different traditional Chinese medicine, more traditional Chinese medicine needs to be developed to better cope with different types of symptoms. SUMMARY

[0005] In view of the deficiencies in the prior art, the purpose of the present application is to provide a traditional Chinese medicine composition for invigorating the spleen and increasing muscle, a pharmaceutical preparation and a preparation method thereof. The traditional Chinese medicine composition and the pharmaceutical preparation have the effects of tonifying the spleen and benefiting qi, eliminating food and dampness, and harmonizing the center and increasing muscle, and can be used for sarcopenia, improving symptoms such as weak spleen and stomach, decreased muscle strength, and deficiency of qi and blood.

[0006] To achieve this purpose, the present application adopts the following technical solutions:

[0007] In the first aspect, the present application provides a traditional Chinese medicine composition for invigorating the spleen and increasing muscle, which comprises the following components:

[0008] Yam, atractylodes, raw dried ginseng, lentil and malt.

[0009] In some preferred embodiments of the present application, the traditional Chinese medicine composition comprises the following components by weight:

[0010] Shanyao 27-32 parts, Baizhu 12-17 parts, Shengsaishen 7-12 parts, Flat bean 12-17 parts and Malt 12-17 parts.

[0011] The weight parts of Shanyao can be any value in the range of 27-32 parts, for example, can be 27 parts, 27.2 parts, 27.5 parts, 27.8 parts, 28 parts, 28.2 parts, 28.5 parts, 28.8 parts, 29 parts, 29.2 parts, 29.5 parts, 29.8 parts, 30 parts, 30.2 parts, 30.5 parts, 30.8 parts, 31 parts, 31.2 parts, 31.5 parts, 31.8 parts or 32 parts, etc.

[0012] The weight parts of Baizhu can be any value in the range of 12-17 parts, for example, can be 12 parts, 12.2 parts, 12.5 parts, 12.8 parts, 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts or 17 parts, etc.

[0013] The weight parts of Shengsaishen can be any value in the range of 7-12 parts, for example, can be 7 parts, 7.2 parts, 7.5 parts, 7.8 parts, 8 parts, 8.2 parts, 8.5 parts, 8.8 parts, 9 parts, 9.2 parts, 9.5 parts, 9.8 parts, 10 parts, 10.2 parts, 10.5 parts, 10.8 parts, 11 parts, 11.2 parts, 11.5 parts, 11.8 parts or 12 parts, etc.

[0014] The weight parts of Flat bean can be any value in the range of 12-17 parts, for example, can be 12 parts, 12.2 parts, 12.5 parts, 12.8 parts, 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts or 17 parts, etc.

[0015] The weight parts of Malt can be any value in the range of 12-17 parts, for example, can be 12 parts, 12.2 parts, 12.5 parts, 12.8 parts, 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts or 17 parts, etc.

[0016] In some preferred embodiments of the present application, the traditional Chinese medicine composition comprises the following components in the weight parts:

[0017] Shanyao 30 parts, Baizhu 15 parts, Shengsaishen 10 parts, Flat bean 15 parts and Malt 15 parts.

[0018] The Chinese medicine composition provided by the present application has the following effects of each component:

[0019] Shanyao: tonifying the spleen and lung, and nourishing the kidney and yin;

[0020] Baizhu: strengthening the spleen and benefiting qi, and drying dampness and promoting water excretion;

[0021] Shengsaishen: greatly tonifying the primordial qi, and benefiting the lung and generating saliva;

[0022] Fangdou: strengthening the spleen and transforming dampness, and harmonizing the center and digesting food;

[0023] Maiya: digesting food and harmonizing the center, and strengthening the spleen and promoting appetite.

[0024] The compatibility mechanism of each component is as follows:

[0025] The monarch drug: Shanyao. It tonifies the spleen and kidney, and nourishes yin and muscle, and provides a material basis for muscle growth. It is aimed at the core pathogenesis of "spleen deficiency and muscle weakness". It promotes nutrient absorption and muscle synthesis by strengthening the spleen, which embodies the theory of "spleen governing muscles".

[0026] The ministerial drugs: Baizhu and Shengsaishen. Baizhu strengthens the spleen and dries dampness, and helps Shanyao to enhance the spleen transportation function, preventing water and dampness from stagnating and hindering nutrient absorption. Shengsaishen greatly tonifies the primordial qi, and enhances the body's synthetic metabolism, directly promoting muscle protein synthesis. Baizhu and Shanyao form a "spleen-strengthening and dampness-transforming" synergy, and Shengsaishen tonifies qi to promote energy conversion. Both of them are ministerial drugs, and strengthen the efficacy of the monarch drug.

[0027] The auxiliary drugs: Fangdou and Maiya. Fangdou strengthens the spleen and transforms dampness, and harmonizes the center and digests food, preventing abdominal distension and poor appetite caused by excessive Shanyao and Baizhu. Maiya digests and removes stagnation, promotes carbohydrate decomposition, improves energy supply, and also regulates liver and spleen qi, and harmonizes the spleen and stomach. Both of them help the monarch and ministerial drugs to absorb and utilize, and prevent the ministerial drugs from being greasy and hindering the stomach, embodying the compatibility wisdom of "tonifying and unblocking".

[0028] The messenger drug: Maiya. Digestive herbs are mild in nature, and can guide other drugs into the spleen and stomach meridians, ensuring that the drug force is concentrated in the middle jiao (digestive system and muscle metabolism). Maiya has a dual role as "auxiliary" and "messenger", regulating the nature of the whole prescription, and avoiding the disadvantages of warming and drying or being greasy.

[0029] The Chinese medicine composition provided by the present application is a food and medicine homologous component, which is safe and suitable for long-term use. It not only increases muscle, but also simultaneously improves symptoms such as indigestion, fatigue, and low metabolism, embodying the advantage of "treating different diseases with the same method". This prescription strengthens the spleen and increases muscle. With Shanyao and Baizhu as the core, it strengthens the spleen and stomach transportation, improves nutrient absorption from the source, solves the fundamental problem of "spleen deficiency and muscle weakness", and avoids relying solely on exogenous nutritional supplements. Shanyao and Shengsaishen greatly tonify qi and yin, Baizhu strengthens the spleen and dries dampness, Fangdou transforms dampness, and Maiya digests food, preventing the stagnation of tonifying drugs, ensuring the absorption of drug efficacy, and realizing the combination of tonifying and unblocking.

[0030] The traditional Chinese medicine composition provided by the present application has the effects of tonifying the spleen and replenishing qi, eliminating food and dampness, and harmonizing the center and increasing muscle, and is suitable for old-age hypomuscular syndrome (spleen qi deficiency syndrome), and can improve spleen and stomach weakness, muscle strength decline, and deficiency of qi and blood.

[0031] In a second aspect, the present application provides a spleen-tonifying and muscle-increasing pharmaceutical preparation, which comprises effective components of the traditional Chinese medicine composition according to the first aspect.

[0032] In some embodiments of the present application, the pharmaceutical preparation further comprises excipients.

[0033] In some embodiments of the present application, the dosage form of the pharmaceutical preparation is granules, tablets, capsules, or decoction.

[0034] In a third aspect, the present application provides a preparation method of the pharmaceutical preparation according to the second aspect, which comprises the following steps:

[0035] (1) water extraction of the traditional Chinese medicine composition according to the first aspect to obtain a water extract;

[0036] (2) preparation of the water extract into the dosage form of the pharmaceutical preparation.

[0037] In some embodiments of the present application, the step of water extraction comprises:

[0038] The traditional Chinese medicine composition is decocted with water for at least 2 times (for example, 2 times, 3 times, 4 times, or 5 times, etc.), each time for 1-4 h (for example, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, or 4 h, etc.), and each time after decoction, the filtrate is collected to obtain the water extract.

[0039] In some embodiments of the present application, in the step of water extraction, the total mass of water is 18-35 times the mass of the traditional Chinese medicine composition; for example, it can be 18 times, 19 times, 20 times, 22 times, 23 times, 25 times, 26 times, 28 times, 30 times, 32 times, 33 times, or 35 times, etc.

[0040] In some embodiments of the present application, in the water extraction step, the decocting is performed twice, in the first decocting, the water is 10-20 times (for example, 10 times, 12 times, 13 times, 15 times, 16 times, 18 times or 20 times, etc.) of the mass of the traditional Chinese medicine composition, and the decocting time is 2-3 hours (for example, 2 hours, 2.2 hours, 2.3 hours, 2.5 hours, 2.6 hours, 2.8 hours or 3 hours, etc.); in the second decocting, the water is 8-15 times (for example, 8 times, 9 times, 10 times, 11 times, 12 times, 13 times, 14 times or 15 times, etc.) of the mass of the traditional Chinese medicine composition, and the decocting time is 1-2 hours (for example, 1 hour, 1.2 hours, 1.3 hours, 1.5 hours, 1.6 hours, 1.8 hours or 2 hours, etc.).

[0041] Compared with the prior art, the present application has the following beneficial effects:

[0042] The present application adopts scientific compatibility of yam, atractylodes, raw dried ginseng, lentil and malt to obtain the traditional Chinese medicine composition and the pharmaceutical preparation, which has the effects of tonifying the spleen and replenishing qi, eliminating dampness and food, and harmonizing the middle and increasing muscle, and can be used for old-age hypomuscular syndrome, and can improve the symptoms of weak spleen and stomach, decreased muscle strength, and insufficient qi and blood. BRIEF DESCRIPTION OF DRAWINGS

[0043] Figure 1 The appearance and signs of each group of mice before taking materials are shown in the photographs;

[0044] Figure 2A The daily food intake change curve of each group of mice after different administration times is shown in the graph;

[0045] Figure 2B The body weight change curve of each group of mice after different administration times is shown in the graph;

[0046] Figure 2C The body weight histogram of each group of mice at the end of the experiment is shown in the graph;

[0047] Figure 3 The fecal water content histogram of each group of mice on the last day of the experiment is shown in the graph;

[0048] Figure 4A The liver organ coefficient histogram of each group of mice is shown in the graph;

[0049] Figure 4B The spleen organ coefficient histogram of each group of mice is shown in the graph;

[0050] Figure 4C The double kidney organ coefficient histogram of each group of mice is shown in the graph;

[0051] Figure 5A The serum ALT content histogram of each group of mice is shown in the graph;

[0052] Figure 5BA column chart of serum AST content of mice in each group;

[0053] Figure 5C A column chart of serum ALP content of mice in each group;

[0054] Figure 6A A column chart of serum D-xylose content of mice in each group;

[0055] Figure 6B A column chart of serum IL-6 content of mice in each group;

[0056] Figure 6C A column chart of serum IL-10 content of mice in each group;

[0057] Figure 7A A HE staining chart of liver tissue of mice in each group;

[0058] Figure 7B A HE staining chart of spleen tissue of mice in each group;

[0059] Figure 7C A HE staining chart of left kidney tissue of mice in each group. DETAILED DESCRIPTION

[0060] The technical solutions of the present application will be further described below in combination with the drawings and through specific embodiments. It should be understood by those skilled in the art that the specific embodiments are only used to help understand the present application and should not be regarded as specific limitations to the present application.

[0061] Embodiment 1

[0062] The present embodiment provides a traditional Chinese medicine composition for invigorating the spleen and increasing muscle, which comprises the following components in parts by weight:

[0063] 27 parts of yam, 17 parts of white atractylodes, 7 parts of raw ginseng, 17 parts of lentil and 12 parts of malt.

[0064] The present embodiment also provides a soup for invigorating the spleen and increasing muscle, and the preparation method is as follows:

[0065] The medicinal materials are weighed according to the proportion of the traditional Chinese medicine composition, and then boiled twice with water. For the first boiling, 20 times the mass of the medicinal materials is added with water, and boiled for 3 hours. For the second boiling, 15 times the mass of the medicinal materials is added with water, and boiled for 2 hours. After each boiling, the filtrate is collected and concentrated under reduced pressure to obtain the soup for invigorating the spleen and increasing muscle.

[0066] Embodiment 2

[0067] The present embodiment provides a traditional Chinese medicine composition for invigorating the spleen and increasing muscle, which comprises the following components in parts by weight:

[0068] 32 parts of yam, 12 parts of white atractylodes, 12 parts of raw ginseng, 12 parts of lentil and 17 parts of malt.

[0069] The embodiment also provides a soup for invigorating the spleen and increasing muscle, and a preparation method thereof is as follows:

[0070] The medicinal materials are weighed according to the proportion of the traditional Chinese medicine composition, and are boiled twice with water. In the first boiling, 10 times the mass of the medicinal materials is added, and boiled for 2 hours. In the second boiling, 8 times the mass of the medicinal materials is added, and boiled for 1 hour. After each boiling, the filtrate is collected, and concentrated under reduced pressure to obtain the soup for invigorating the spleen and increasing muscle.

[0071] Embodiment 3

[0072] The embodiment provides a traditional Chinese medicine composition for invigorating the spleen and increasing muscle, which comprises the following components in parts by weight:

[0073] 30 parts of yam, 15 parts of white atractylodes, 10 parts of dried ginseng, 15 parts of lentil and 15 parts of malt.

[0074] The embodiment also provides a soup for invigorating the spleen and increasing muscle, and a preparation method thereof is as follows:

[0075] The medicinal materials are weighed according to the proportion of the traditional Chinese medicine composition, and are boiled twice with water. In the first boiling, 15 times the mass of the medicinal materials is added, and boiled for 2.5 hours. In the second boiling, 10 times the mass of the medicinal materials is added, and boiled for 1.5 hours. After each boiling, the filtrate is collected, and concentrated under reduced pressure to 3.5 g of the traditional Chinese medicine composition per mL to obtain the soup for invigorating the spleen and increasing muscle, which is stored at 4°C for standby.

[0076] Therapeutic effect test

[0077] The soup for invigorating the spleen and increasing muscle prepared in Embodiment 3 is used for animal experiments to confirm the therapeutic effect on the mouse liver stagnation and spleen deficiency model.

[0078] 1. Test materials

[0079] 1.1. Test animals

[0080] SPF level male ICR mice aged 5-7 weeks are used for the test, which are purchased from Sibeifu (Suzhou) Biotechnology Co., Ltd. with production license number SCXK (Su) 2022-0006 and qualification certificate number A202411260116. The feeding environment conditions are as follows: the standard of feeding environment conditions is referred to the national standard of the People's Republic of China GB14925-2010; the temperature is 20-26°C (the daily temperature difference is less than or equal to 4°C); the relative humidity is 40-70%; the pressure in the feeding room is greater than or equal to 10 Pa; and the day and night alternation is 12 / 12 hours.

[0081] 1.2. Test reagents

[0082] The spleen-strengthening and muscle-increasing decoction prepared in Example 3 was diluted with sterile water for injection to 0.702 g / mL, 1.404 g / mL, and 2.808 g / mL, respectively. The concentration refers to the mass of the corresponding traditional Chinese medicine composition contained in 1 mL of the solution.

[0083] Rhubarb decoction, specification 0.5 g of rhubarb / mL, stored at 4°C, decocted, concentrated, and provided by Jiangsu Yongjian Pharmaceutical Technology Co., Ltd.

[0084] Shenling Baizhu Powder, specification 12 g / bag, batch number Z11020755, stored in a sealed manner, and produced by Beijing Tong Ren Tang Co., Ltd. Tong Ren Tang Pharmaceutical Factory. Preparation method: diluted with sterile water for injection to 2.703 g / mL.

[0085] Sterile water for injection, specification 500 mL / bottle, batch number 240621, stored in a sealed manner, and produced by Shaanxi Shengao Animal Pharmaceutical Co., Ltd.; product approval number Veterinary Drug 270071791.

[0086] General tissue fixative, specification 500 mL / bottle, batch number GP24103081634, stored at room temperature, and produced by Wuhan Saiver Biological Technology Co., Ltd.

[0087] 1.3. Instruments

[0088] Pipette gun, produced by Eppendorf, model 200 μL;

[0089] Electronic balance, produced by Shanghai Yaoxin Electronic Technology Co., Ltd., model LQ-C12001;

[0090] Tabletop high-speed refrigerated microcentrifuge, produced by Thermo, model Sorvall Legand Micro 17R;

[0091] Mini mixing and centrifuging integrated machine, produced by LABGIC, model L-CM-MINI;

[0092] Enzyme label instrument, produced by Thermo, model VARIOSKAN LUX;

[0093] Dehydrator, produced by DIAPATH, model Donatello;

[0094] Embedding machine, produced by Wuhan Junjie Electronic Co., Ltd., model JB-P5;

[0095] Pathological sectioning machine, produced by Shanghai Leica Instruments Co., Ltd., model RM2016;

[0096] Tissue slicer, manufacturer Zhejiang Jinhua City Codi Instrument and Equipment Co., Ltd., model KD-P;

[0097] Oven, manufacturer Tianjin Leborui Instrument and Equipment Co., Ltd., model GFL-230;

[0098] Orthoplan optical microscope, manufacturer Nikon, model Nikon Eclipse E100;

[0099] Imaging system, manufacturer Nikon, model NIKON DS-U3;

[0100] Orthoplan white light photographing microscope, manufacturer Nikon, model Eclipse Ci-L;

[0101] Small animal treadmill, manufacturer Nanjing Calvin Biotechnology Co., Ltd., model KW-PT.

[0102] 2. Test method

[0103] 2.1. Modeling, grouping and administration

[0104] After the mice were adaptively fed with normal diet for 1 week, they were randomly divided into 6 groups, namely, blank group, model group, prescription low-dose group, prescription medium-dose group, prescription high-dose group and Shenling Baizhu Powder group, 8 mice in each group.

[0105] The blank group was normally bred, and the other groups of mice were modeled for 4 weeks by the combined method of “rhubarb bitter cold purgation + improper diet + overwork” except the blank group to prepare the mouse liver stagnation and spleen deficiency model.

[0106] Rhubarb bitter cold purgation: 0.2 mL / 10 g·d of 0.5 g / mL rhubarb decoction was given by gavage;

[0107] Overwork: the mice were placed in the treadmill at regular intervals every day to make them run to exhaustion;

[0108] Improper diet: the diet of the mice was controlled every day, and they were fed alternately every other day (one day of fasting without water and one day of sufficient food supply, alternating in cycles), so that the mice were in a state of irregular hunger and satiety.

[0109] After the modeling was successful, the mice in each group were given medicine by gavage twice a day (once in the morning and once in the evening), for 27 consecutive days, and 0.2 mL / 10 g·d of 0.5 g / mL rhubarb decoction was given by gavage at noon every day.

[0110] The types and doses of medicine for the mice in each group were as follows:

[0111] Blank group and model group: normal saline, the volume of medicine was 100 μL / 10 g;

[0112] Prescription low-dose group: dilution of the decoction prepared in Example 3 with a concentration of 0.702 g / mL, administration dose 7.02 g / kg, administration volume 100 μL / 10 g;

[0113] Prescription medium-dose group: dilution of the decoction prepared in Example 3 with a concentration of 1.404 g / mL, administration dose 14.04 g / kg, administration volume 100 μL / 10 g;

[0114] Prescription high-dose group: dilution of the decoction prepared in Example 3 with a concentration of 2.808 g / mL, administration dose 28.08 g / kg, administration volume 100 μL / 10 g;

[0115] Shenlingbaishu group: dilution of Shenlingbaishu with a concentration of 2.703 g / mL, administration dose 27.03 g / kg, administration volume 100 μL / 10 g.

[0116] 2.2. Detection index and method

[0117] 2.2.1. Observation of appearance and behavior index of mice

[0118] Appearance and behavior index includes: listlessness, hunchback and squinting, shivering and huddling, coat condition, startle response, loose stool and diarrhea, etc.

[0119] 2.2.2. Daily food intake statistics

[0120] 2.2.3. Body weight

[0121] Daily weighing, statistics of body weight change of mice.

[0122] 2.2.4. Fecal water content detection

[0123] On the last day of the experiment, the mice were fed in metabolic cages, and the 24 h feces of the mice were collected, dried, and the water content was calculated according to the change in mass of the feces before and after drying.

[0124] 2.2.5. Organ coefficient: liver, spleen, kidney

[0125] Determine the body weight of the mice, and after the mice are sacrificed, take the liver, spleen and both kidneys, weigh them, and calculate the organ coefficient according to the following formula: organ coefficient = organ weight / body weight x 100%.

[0126] 2.2.6. HE staining: liver, spleen, kidney

[0127] (1) Paraffin section preparation

[0128] 1) Sample collection: After the experiment, the mice were sacrificed, and the liver, spleen, and kidney tissues were collected and fixed in fixative for more than 24 hours. The tissues were removed from the fixative and trimmed with a scalpel in a fume hood. The trimmed tissues and corresponding labels were placed in a dehydration box.

[0129] 2) Dehydration and wax immersion: The dehydration box was placed in a dehydration machine for gradient alcohol dehydration. The sequence was as follows: 75% alcohol for 4 hours, 85% alcohol for 2 hours, 90% alcohol for 2 hours, 95% alcohol for 1 hour, anhydrous ethanol I for 30 minutes, anhydrous ethanol II for 30 minutes, benzyl alcohol for 10 minutes, xylene I for 10 minutes, xylene II for 10 minutes, 65°C melted paraffin I for 1 hour, 65°C melted paraffin II for 1 hour, and 65°C melted paraffin III for 1 hour.

[0130] 3) Embedding: The wax-embedded tissues were placed in an embedding machine for embedding. The melted wax was first placed in an embedding frame, and the tissues were removed from the dehydration box before the wax solidified. The tissues were placed in the embedding frame according to the requirements of the embedding surface and labeled accordingly. The embedding frame was cooled on a -20°C freezing stage, and the wax block was removed from the embedding frame after the wax solidified and trimmed.

[0131] 4) Sectioning: The trimmed wax block was placed in a -20°C freezing stage, and the cooled wax block was sectioned on a paraffin microtome with a thickness of 4μm. The sections were floated on a 40°C water bath on a section flattener to flatten the tissues, and the glass slides were used to retrieve the tissues. The slides were baked in a 60°C oven to dry the wax.

[0132] (2) Staining

[0133] 1) Paraffin section dehydration to water: The sections were sequentially placed in xylene I for 20 minutes, xylene II for 20 minutes, anhydrous ethanol I for 5 minutes, anhydrous ethanol II for 5 minutes, 75% alcohol for 5 minutes, and tap water.

[0134] 2) Hematoxylin staining: The dehydrated sections were placed in hematoxylin staining solution for 5 minutes, washed with tap water, differentiated with differentiation solution, washed with tap water, and returned to blue with blue return solution, and then washed with running water.

[0135] 3) Eosin staining: The hematoxylin-stained sections were sequentially placed in 85% and 95% alcohol for 5 minutes each, and then placed in eosin staining solution for 5 minutes.

[0136] 4) Dehydration and mounting: The eosin-stained sections were sequentially placed in anhydrous ethanol I for 5 minutes, anhydrous ethanol II for 5 minutes, anhydrous ethanol III for 5 minutes, xylene I for 5 minutes, xylene II for 5 minutes, and transparent neutral balsam for mounting.

[0137] 5) Microscopic examination, image acquisition and analysis.

[0138] 2.2.7. Detection of serum AST, ALT and ALP contents

[0139] After sampling, set the corresponding parameters on the full-automatic biochemical analyzer and then load the sample for detection.

[0140] 2.2.8. Detection of serum IL-6 and IL-10 contents

[0141] The IL-6 and IL-10 contents in the serum were detected according to the detection steps of the mouse IL-6 and IL-10 detection kits.

[0142] 2.2.9. Detection of serum D-xylose content

[0143] The serum D-xylose content was detected according to the detection steps of the D-xylose content detection kit.

[0144] 2.3. Data analysis and processing

[0145] GraphPad 9.5.0 and SPSS26 software were used for drawing and statistical analysis, the data were expressed as mean ± SEM, statistical analysis was performed using one-way ANOVA, and significant differences between two groups were analyzed by t-test, P<0.05 indicating statistical significance.

[0146] 3. Test results

[0147] 3.1. Influence of the prescription of the application on the appearance, food intake, body weight and fecal water content of liver stagnation and spleen deficiency model mice

[0148] Figure 1 The appearance of the mice in each group before sampling was photographed. Figure 2A The daily food intake change curve of the mice in each group after different administration times was plotted, Figure 2B The body weight change curve of the mice in each group after different administration times was plotted, Figure 2C The body weight histogram of the mice in each group at the end of the experiment was plotted. Figure 3 The fecal water content histogram of the mice in each group on the last day of the experiment was plotted. Compared with the blank group, *** p<0.001; compared with the model group, # p<0.05, ## p<0.01, ### p<0.001; compared with the Shenling Baizhu Powder group, @ p<0.05.

[0149] From Figures 1-3 it can be seen that, compared with the blank group, the model group of mice showed symptoms of mental fatigue, hunched back, eye squinting, cold and huddling, rough and disordered fur, nervous sensitivity, and loose stool and diarrhea, and after treatment with the prescription of the application and the Shenling Baizhu Powder, the above symptoms were significantly improved.

[0150] Compared with the blank group, the food intake and body weight of the mice in the model group decreased (P<0.001); compared with the model group, the food intake and body weight of the mice in each dose group of the prescription and the positive drug Shenling Baizhu San group increased (P<0.001).

[0151] Compared with the blank group, the fecal water content of mice in the model group was significantly increased (P<0.001); compared with the model group, the fecal water content of mice in each dose group of the prescription and the positive drug Shenling Baizhu San group was significantly decreased (P<0.05, P<0.01, P<0.001), and the fecal water content of mice in the medium dose group of the prescription was significantly lower than that in the Shenling Baizhu San group (P<0.05).

[0152] 3.2. Effects of the present invention's prescription on organ coefficients in mice with liver depression and spleen deficiency model

[0153] Figure 4A is the bar graph of liver organ coefficient of each group of mice, Figure 4B is the spleen organ coefficient bar graph of each group of mice, Figure 4C The figure is the histogram of the kidney organ coefficients of each group of mice. Compared with the blank group, *** p<0.001; compared with the model group, # p<0.05, ### p<0.001.

[0154] from Figures 4A-4C It can be seen that compared with the blank group, the liver organ coefficient of the model group mice increased significantly (P < 0.001), the spleen organ coefficient decreased significantly (P < 0.001), and there was no significant difference in the kidney organ coefficient. Compared with the model group, the liver organ coefficient of the mice in each dosage group of the prescription and the positive drug Shenling Baizhu powder group decreased significantly (P < 0.001), the spleen organ coefficient increased significantly (P < 0.05), and there was no significant difference in the kidney organ coefficient. This is consistent with the pathological manifestations of "liver depression and spleen deficiency" in traditional Chinese medicine (liver qi stagnation may cause an increase in the metabolic load of the liver, and spleen deficiency is reflected as hyposplenia), and the kidney organ coefficient did not change, indicating that the model had no direct effect on the kidney. The prescription of the present invention and the positive drug Shenling Baizhu powder can effectively alleviate the liver and spleen damage of the spleen deficiency model mice, soothe the liver and relieve depression, and improve spleen deficiency.

[0155] 3.3. Effects of the present invention's prescription on serum biochemical indices in mice with liver depression and spleen deficiency model

[0156] Figure 5A is a bar graph of serum ALT levels in each group of mice. Figure 5B is the bar graph of serum AST content in each group of mice. Figure 5C is a bar graph of serum ALP content in each group of mice. Figure 6A is a bar graph of serum D-xylose content in each group of mice. Figure 6Bis a bar graph of serum IL-6 levels in each group of mice. Figure 6C The bar graph shows the serum IL-10 content of each group of mice. Compared with the blank group, * p<0.05, ** p<0.01, *** p<0.001; compared with the model group, # p<0.05, ## p<0.01, ### p<0.001; compared with the Shenling Baizhu powder group, @ p<0.05.

[0157] Elevated levels of ALT (alanine aminotransferase), AST (aspartate aminotransferase), and ALP (alkaline phosphatase) indicate liver damage; decreased D-xylose levels suggest intestinal absorption dysfunction (common in spleen deficiency models). IL-6 (interleukin-6) and IL-10 (interleukin-10) are inflammatory cytokines. Elevated serum IL-6 levels or decreased IL-10 levels indicate an inflammatory response.

[0158] from Figures 5A-5C It can be seen that compared with the blank group, the ALT, AST, and ALP levels in the serum of the mice in the model group were significantly increased (P<0.01, P<0.001); compared with the model group, the ALT, AST, and ALP levels in the serum of the mice in each dose group of the prescription and the positive drug Shenling Baizhu San group were significantly decreased (P<0.01, P<0.001). This shows that the prescription of the present invention and the positive drug Shenling Baizhu San can effectively improve liver damage caused by liver depression and spleen deficiency.

[0159] from Figures 6A-6C It can be seen that compared with the blank group, the IL-6 content in the serum of the model group mice was significantly increased (P<0.001), and the D-xylose (P<0.05) and IL-10 (P<0.001) contents were significantly reduced. Compared with the model group, the IL-6 content in the serum of the mice in each dosage group of the prescription and the positive drug Shenling Baizhu San group was significantly reduced (P<0.001), and the D-xylose (P<0.05, P<0.01) and IL-10 content were significantly increased (P<0.001). This shows that the prescription of the present invention and the positive drug Shenling Baizhu San can effectively improve intestinal function and alleviate the inflammatory imbalance caused by spleen deficiency.

[0160] 3.4. Effects of the Prescription of the Present Invention on Organ Tissues in Mice Models of Liver Depression and Spleen Deficiency

[0161] Figure 7A HE staining images of liver tissues of mice in each group (200×). Figure 7B HE staining images of spleen tissues of mice in each group (200×). Figure 7CHE staining images (200x) of the left kidney tissues of mice in each group. A: blank group, B: model group, C: prescription low-dose group, D: prescription middle-dose group, E: prescription high-dose group, and F: Shenling Baizhu Powder group.

[0162] As shown in Table 1, compared with the blank group, the model group showed a significant increase in the number of samples with inflammatory cell infiltration, and a significant decrease in the number of samples with normal liver tissue. Compared with the model group, the prescription low-dose group showed a significant decrease in the number of samples with inflammatory cell infiltration, and a significant increase in the number of samples with hepatocyte edema. The prescription middle-dose group showed no significant inflammatory cell infiltration. The prescription high-dose group showed a significant decrease in the number of samples with inflammatory cell infiltration, and a significant increase in the number of samples with hepatocyte edema. The positive drug Shenling Baizhu Powder group showed a significant decrease in the number of samples with inflammatory cell infiltration, and a significant increase in the number of samples with hepatocyte edema. Figure 7A As shown in Table 2, compared with the blank group, the model group showed a significant increase in the number of samples with inflammatory cell infiltration in the red pulp, and a significant increase in the number of granulocytes. Compared with the model group, the prescription low-dose group showed a significant decrease in the number of samples with granulocytes in the red pulp. The prescription middle-dose group showed a significant decrease in the number of granulocytes in the red pulp. The prescription high-dose group showed a significant decrease in the number of granulocytes in the red pulp, and a significant increase in the number of samples with splenic sinus expansion. The positive drug Shenling Baizhu Powder group showed a significant decrease in the number of granulocytes in the red pulp.

[0163] Figure 7B As shown in Table 3, compared with the blank group, the model group showed a significant increase in the number of samples with inflammatory cell infiltration around the renal pelvis and blood vessels, a significant increase in the number of samples with interstitial blood vessel stasis, and a significant increase in the number of samples with renal tubular expansion. Compared with the model group, the prescription low-dose group, the prescription high-dose group, and the positive drug Shenling Baizhu Powder group showed no significant difference. The prescription middle-dose group showed a significant increase in the number of samples with interstitial blood vessel stasis.

[0164] As shown in Table 4, compared with the blank group, the model group showed a significant increase in the number of samples with inflammatory cell infiltration around the renal pelvis and blood vessels, a significant increase in the number of samples with interstitial blood vessel stasis, and a significant increase in the number of samples with renal tubular expansion. Compared with the model group, the prescription low-dose group, the prescription high-dose group, and the positive drug Shenling Baizhu Powder group showed no significant difference. The prescription middle-dose group showed a significant increase in the number of samples with interstitial blood vessel stasis. Figure 7C As shown in Table 5, compared with the blank group, the model group showed a significant increase in the number of samples with inflammatory cell infiltration around the renal pelvis and blood vessels, a significant increase in the number of samples with interstitial blood vessel stasis, and a significant increase in the number of samples with renal tubular expansion. Compared with the model group, the prescription low-dose group, the prescription high-dose group, and the positive drug Shenling Baizhu Powder group showed no significant difference. The prescription middle-dose group showed a significant increase in the number of samples with interstitial blood vessel stasis.

[0165] ​In conclusion, the liver damage of the liver stagnation and spleen deficiency model mice is reduced, the body weight is increased, the food intake is increased, the fecal water content is reduced, and the inflammation is reduced after the intervention of the prescription and the positive medicine Shengling Baizhu Powder. The efficacy of the prescription is flat with the positive medicine Shengling Baizhu Powder.

[0166] The above description is merely that of a specific implementation of the disclosure, which enables those skilled in the art to understand or implement the disclosure. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the disclosure. Therefore, the disclosure will not be limited to these embodiments described herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A Chinese medicine composition for strengthening the spleen and increasing muscle mass, characterized in that: The Chinese medicine composition comprises the following components: Chinese yam, Atractylodes macrocephala, raw sun-dried ginseng, lentils and malt.

2. The Chinese medicine composition according to claim 1, characterized in that The Chinese medicine composition comprises the following components in parts by weight: 27-32 parts of Chinese yam, 12-17 parts of white atractylodes, 7-12 parts of raw sun-dried ginseng, 12-17 parts of lentils and 12-17 parts of malt.

3. The Chinese medicine composition according to claim 1 or 2, characterized in that The Chinese medicine composition comprises the following components in parts by weight: 30 parts of Chinese yam, 15 parts of Atractylodes macrocephala, 10 parts of sun-dried ginseng, 15 parts of lentils and 15 parts of malt.

4. A pharmaceutical preparation for strengthening the spleen and increasing muscle mass, characterized in that: The pharmaceutical preparation comprises the active ingredient of the traditional Chinese medicine composition according to any one of claims 1 to 3.

5. The pharmaceutical preparation according to claim 4, characterized in that The pharmaceutical preparation further includes an excipient.

6. The pharmaceutical preparation according to claim 4 or 5, characterized in that The dosage form of the pharmaceutical preparation is granules, tablets, capsules or decoctions.

7. A method for preparing the pharmaceutical preparation according to any one of claims 4 to 6, characterized in that: The preparation method comprises the following steps: (1) extracting the Chinese medicine composition according to any one of claims 1 to 3 with water to obtain a water extract; (2) preparing the aqueous extract into the dosage form of the pharmaceutical preparation.

8. The preparation method according to claim 7, characterized in that The step of water extraction comprises: The traditional Chinese medicine composition is decocted with water for at least 2 times, each time for 1-4 hours, filtered after each decoction, and the filtrates are combined to obtain the water extract.

9. The preparation method according to claim 7 or 8, characterized in that In the water extraction step, the total mass of water is 18-35 times the mass of the traditional Chinese medicine composition.

10. The preparation method according to claim 8, characterized in that In the water extraction step, the decoction is performed twice. In the first decoction, the mass of water is 10-20 times the mass of the traditional Chinese medicine composition, and the decoction time is 2-3 hours; in the second decoction, the mass of water is 8-15 times the mass of the traditional Chinese medicine composition, and the decoction time is 1-2 hours.

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