Tangzhou ginseng soup composition as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a radix codonopsis, poria cocos and ginseng decoction composition and a preparation method and application thereof.The radix codonopsis, poria cocos and ginseng decoction composition is prepared from, by mass, 9-15 parts of tabasheer, 9-15 parts of poria cocos, 9-15 parts of radix codonopsis, 10-20 parts of ulva lactuca, 10-20 parts of seaweed, 3-9 parts of hawthorn and 3-9 parts of radix bupleuri; the preparation method comprises the following steps: weighing the components according to the formula, adding water for soaking, decocting, filtering, and collecting primary filtrate; taking filter residues, adding water again, decocting, then filtering, and collecting secondary filtrate; and merging the two filtrates, and uniformly mixing to obtain the radix aconiti, poria cocos and ginseng soup. Compared with the prior art, the traditional Chinese medicine composition and the traditional Chinese medicine preparation for MASLD solve the problems that in the prior art, treatment medicine has obvious untoward effects and is single in curative effect, and meanwhile, a traditional Chinese medicine composition and a traditional Chinese medicine preparation for MASLD are lacked. According to the scheme, a personalized traditional Chinese medicine intervention strategy is provided for MASLD, especially for patients with the phlegm-dampness and blood-stasis-heat syndrome fatty liver.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a Tianfushen Decoction composition, its preparation method, and its application. Background Technology

[0002] Metabolic dysfunction-associated steatotic liver disease (MASLD) is a multi-system metabolic stress-induced liver injury. Epidemiological studies show that the prevalence of MASLD is increasing year by year, and it has become the leading cause of chronic liver disease worldwide, posing a significant impact on human health and imposing a huge economic burden on public health systems. Notably, the patient population with MASLD is becoming increasingly younger, indicating that more and more patients may suffer from MASLD and its related complications for a long time.

[0003] In response to the severe challenges posed by this major health problem, the nation has proposed clear health management strategies. However, current treatments do not offer satisfactory intervention strategies. In recent years, some progress has been made in drug development, including GLP-1 receptor agonists (such as smegglutide), farnesol X receptor agonists (such as obeticholic acid), and PPARα / δ dual agonists (such as Elafibranor, Resmetirom, Centicriviroc, and selonsertib). However, in clinical practice, these drugs have all exhibited limitations to varying degrees, specifically including: significant adverse reactions, limited therapeutic efficacy, and an inability to comprehensively address the systemic metabolic problems of MASLD.

[0004] Therefore, exploring innovative treatment options and seeking to shift from controlling a single cause to multi-mechanism combined intervention strategies may provide more assistance for the development of drugs for the treatment of MASLD. In this context, traditional Chinese medicine, with its systemic advantages and low toxicity, is receiving increasing attention from scholars both domestically and internationally in the prevention and treatment of MASLD.

[0005] In the prior art, CN109157644A discloses a traditional Chinese medicine composition for invigorating the spleen and nourishing the liver and its preparation method; CN106880817A discloses a traditional Chinese medicine composition for treating hepatitis B and its preparation method; CN109248305A discloses a traditional Chinese medicine composition for treating liver fibrosis and its preparation method; CN118320043A discloses a composition for invigorating the spleen, clearing the meridians, and reducing turbidity for treating non-alcoholic steatohepatitis (NASH). There have been studies on the prevention and treatment of liver-related diseases using traditional Chinese medicine compositions, but no relevant studies on the prevention and treatment of MASLD have been found so far.

[0006] The advantages of Traditional Chinese Medicine (TCM) lie in early and long-term intervention. However, a challenge in clinical application is that, due to the combined nature of disease and syndrome differentiation, different TCM syndrome types require targeted intervention plans. Therefore, developing clinically effective prescriptions and drugs adapted to different syndrome types can provide personalized TCM intervention strategies for MASLD. Summary of the Invention

[0007] The purpose of this invention is to address at least one of the aforementioned problems by providing a Tianfu Shentang composition, its preparation method, and its application. This addresses the issues of significant adverse reactions and limited efficacy of existing therapeutic drugs, as well as the lack of traditional Chinese medicine compositions and preparations specifically for MASLD. This solution provides a personalized traditional Chinese medicine intervention strategy for MASLD, particularly for patients with fatty liver due to phlegm-dampness and blood stasis.

[0008] The objective of this invention is achieved through the following technical solution:

[0009] From the perspective of Traditional Chinese Medicine (TCM), MASLD falls under the categories of "hypochondriac pain," "accumulation," and "liver stagnation." This disease is often a case of deficiency in the root and excess in the branch: on the basis of spleen qi deficiency, coupled with improper diet, overwork, and emotional imbalance, liver qi stagnation occurs, leading to internal dampness, which, over time, transforms into heat. Ultimately, this dampness combines with multiple pathological factors such as phlegm, turbidity, blood stasis, and toxins to cause the disease. Among these, damp-heat and phlegm-blood stasis are important pathogenic mechanisms in the progression of MASLD and often occur together. Therefore, based on long-term clinical practice, we have developed a medicinal combination called Tianfu Shen Tang for patients with fatty liver syndrome characterized by phlegm-dampness-blood stasis-heat.

[0010] The first aspect of this invention discloses a Tianfushen Decoction composition, comprising the following components in parts by weight:

[0011] 9-15 parts of Tianzhuhuang, 9-15 parts of Fuling, 9-15 parts of Dangshen, 10-20 parts of Shichun, 10-20 parts of Haizao, 3-9 parts of Shanzha, and 3-9 parts of Chaihu.

[0012] Preferably, the Tianfu Shentang composition comprises the following components in parts by weight:

[0013] 9 parts of Tianzhuhuang, 9 parts of Fuling, 9 parts of Dangshen, 10 parts of Shichun, 10 parts of Haizao, 3 parts of Shanzha, and 3 parts of Chaihu;

[0014] or,

[0015] 12 parts of Tianzhuhuang, 12 parts of Fuling, 12 parts of Dangshen, 15 parts of Shichun, 15 parts of Haizao, 6 parts of Shanzha, and 6 parts of Chaihu.

[0016] or,

[0017] 15 parts of Tianzhuhuang (Bamboo Sap), 15 parts of Fuling (Poria), 15 parts of Dangshen (Codonopsis), 20 parts of Shichun (Ulva), 20 parts of Haizao (Sargassum), 9 parts of Shanzha (Crataegus pinnatifida), and 9 parts of Chaihu (Bupleurum chinense).

[0018] Preferably, the Tianfu Shentang composition comprises the following components in parts by weight:

[0019] 12 parts of Tianzhuhuang (Bamboo Sap), 12 parts of Fuling (Poria), 12 parts of Dangshen (Codonopsis), 15 parts of Shichun (Ulva), 15 parts of Haizao (Sargassum), 6 parts of Shanzha (Crataegus pinnatifida), and 6 parts of Chaihu (Bupleurum chinense).

[0020] Preferably, all components in the Tianfushen Decoction composition are raw medicinal materials.

[0021] A second aspect of this invention discloses a method for preparing the Tianfushen Decoction composition as described in any of the above descriptions, comprising the following steps:

[0022] S1: Weigh each component according to the formula, soak in water, boil and filter, and collect the first filtrate.

[0023] S2: Take the filter residue, add water again and boil, then filter and collect the secondary filtrate;

[0024] S3: Combine the two filtrates and mix them well to obtain Tianfu Shentang.

[0025] Preferably, step S1 includes one or more of the following:

[0026] i) The mass of water added should be 6-10 times the total mass of all components in the formula;

[0027] ii) Soaking time is between 20 and 40 minutes;

[0028] iii) The simmering time is between 20 and 40 minutes.

[0029] Preferably, step S2 includes one or both of the following:

[0030] i) The mass of water added should be 6-10 times the mass of the filter cake;

[0031] ii) The simmering time is between 20 and 40 minutes.

[0032] Preferably, the following steps are also included:

[0033] S4: The Tianfu Shentang obtained in step S3 is concentrated under reduced pressure to prepare a concentrated Tianfu Shentang solution of a specified concentration.

[0034] Preferably, step S4 includes one or both of the following:

[0035] i) The vacuum concentration is carried out at 40-60°C;

[0036] ii) The concentration of the Tianfushen Decoction concentrate is 1-4g of raw material per milliliter, wherein the raw material refers to all components in the composition.

[0037] The third aspect of this invention discloses the use of the Tianfushen Decoction composition as described above in the preparation of a medicament for metabolic dysfunction-related fatty liver disease.

[0038] Compared with the prior art, the present invention has the following beneficial effects:

[0039] The Tianfu Shentang composition of this invention, with all the herbs working together, achieves the effects of clearing heat and dampness, resolving phlegm and eliminating blood stasis. The herbs are rationally combined, with no obvious superfluous ingredients, making it well-suited for the prevention and intervention of MASLD phlegm-dampness-blood-stasis-heat syndrome, and possesses promising application prospects.

[0040] The prescription for Tianfu Shen Tang is explained as follows:

[0041] Principal ingredient: Bambusa textilis. As the principal ingredient, Bambusa textilis has the effect of clearing heat and resolving phlegm. It is recorded in "Xiao'er Yaozheng Zhijue" that it is particularly good at treating infantile malnutrition. In the drug combination of this invention, it targets the pathogenesis of internal obstruction of phlegm and dampness and heat disturbing the liver and gallbladder, providing the main treatment direction for the entire prescription.

[0042] Assistant herbs: Poria cocos and Codonopsis pilosula. As assistant herbs, Poria cocos strengthens the spleen and eliminates dampness, calms the mind and soothes the nerves, and can improve the spleen deficiency and water retention, thus helping to eliminate dampness and resolve phlegm; Codonopsis pilosula invigorates qi and strengthens the spleen, enhances the spleen and stomach's digestive function, and assists the main herbs in improving the internal causes of dampness accumulation and phlegm production.

[0043] Adjuvant herbs: Ulva prostrata, Sargassum, and Hawthorn. Ulva prostrata, also known as Guangkunbu, when used together with Sargassum, both have the effects of softening and dispersing nodules, resolving phlegm and removing blood stasis, and are used to resolve symptoms of fat accumulation and phlegm and blood stasis in the liver area; Hawthorn promotes digestion and removes food stagnation, and can improve food stagnation and blood stasis, and help reduce the burden on the liver.

[0044] Guide herb: Bupleurum. As the guide herb, Bupleurum soothes the liver and relieves depression, and promotes the upward movement of clear Qi. It can regulate liver Qi and promote the circulation of Qi and blood, and guide other herbs to the liver and gallbladder, thereby enhancing the overall therapeutic effect of the prescription.

[0045] Clinical observations have shown that this formula not only has a good relieving and therapeutic effect on MASLD, but also shows particularly significant efficacy in patients with fatty liver and stress-induced obesity. This suggests that the formula has a certain therapeutic effect on stress and mood, and its effect is likely mainly due to the heat-clearing, phlegm-resolving, calming, and tranquilizing effects of the herbal pair of Bambusa textilis, Poria cocos, and Codonopsis pilosula. Attached Figure Description

[0046] Figure 1 The effect of Tianfushen Decoction on body weight in MASLD mice in animal model experiments;

[0047] Figure 2 The effect of Tianfushen Decoction on liver weight in MASLD mice in animal model experiments;

[0048] Figure 3The effect of Tianfushen Decoction on blood lipids in MASLD mice in an animal model experiment;

[0049] Figure 4 The effect of Tianfushen Decoction on liver function in MASLD mice in animal model experiments;

[0050] Figure 5 The effect of Tianfushen Decoction on blood glucose in MASLD mice in animal model experiments;

[0051] Figure 6 The effect of Tianfushen Decoction on liver tissue pathology in MASLD mice in animal model experiments;

[0052] Figure 7 The effect of Tianfushen Decoction on the level of oxidative stress in liver tissue of MASLD mice in an animal model experiment. Detailed Implementation

[0053] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0054] Unless otherwise specified, the reagents used in the following description are conventional commercial products, the methods used are known in the art, and any other matters not covered herein are existing technologies.

[0055] Example

[0056] The specific composition and dosage ratio of the herbs in Tianfu Shen Decoction are as follows:

[0057] Bamboo shavings 9-15g, Poria cocos 9-15g, Codonopsis pilosula 9-15g, Ulva lactuca 10-20g, Sargassum fusiforme 10-20g, Crataegus pinnatifida 3-9g, Bupleurum chinense 3-9g.

[0058] The specific preparation method of Tianfushen Decoction is as follows:

[0059] According to the formula, take the raw herbs Tianzhuhuang (Bamboo Sap), Poria cocos, Codonopsis pilosula, Ulva lactuca, Sargassum, Crataegus pinnatifida, and Bupleurum chinense. Add 6-10 times the total mass of each component to water and soak for 20-40 minutes. Then decoct for 20-40 minutes and filter. Add 6-10 times the amount of water to the residue of the raw herbs again and decoct for 20-40 minutes. Filter again. Combine the two filtrates and mix well to obtain Tianfushen Decoction.

[0060] When administering the Poria and Ginseng Decoction to the patient, it should be taken in two divided doses.

[0061] Animal model experiments:

[0062] Male, normal C57BL / 6 mice aged 6-8 weeks were selected. Mice were weighed, their toes clipped, and numbered. They were then randomly grouped into 10 groups of 6 mice each using a random number table. Mice had free access to food and water and were housed for 16 weeks.

[0063] Group 1: Normal control group, fed with ordinary control feed.

[0064] Group 2: Model group, fed a high-fat, high-fructose, and high-cholesterol diet (HFD, Synergistic Bio, XT310). The high-fat, high-fructose, and high-cholesterol diet induced the generation of phlegm and dampness in the body. Long-term modeling led to blood stasis and heat accumulation. This manifested as oily fur, decreased activity, and a distinct urine odor in the mice, indicating the successful establishment of the fatty liver model and the construction of the MASLD mouse model.

[0065] Group 3: Low-dose treatment group of Tianfushen Decoction. Patients were given a high-fat, high-fructose, and high-cholesterol diet. The following herbs were used: 9g of raw Tianzhuhuang (Bamboo Sap), 9g of Fuling (Poria), 9g of Dangshen (Codonopsis), 10g of Shichun (Ulva), 10g of Haizao (Sargassum), 3g of Shanzha (Crataegus pinnatifida), and 3g of Chaihu (Bupleurum). These were soaked in 6 times the amount of water for 40 minutes, decocted for 20 minutes, filtered, and then decocted again in 6 times the amount of water for 20 minutes. The filtrates were combined and mixed, then concentrated under reduced pressure at 40-60℃ until the concentration was 1g of raw material / mL, suitable for animal experiments. The final dosage for experimental mice was 5.07g / kg / d (calculated by gavage based on the conversion rate of human and animal body surface area, referring to "Methodology of Pharmacological Research of Traditional Chinese Medicine").

[0066] Group 4: Medium-dose treatment group with Tianfu Shentang. The patient was fed a high-fat, high-fructose, and high-cholesterol diet. The following herbs were used: 12g of raw Tianzhuhuang (Bamboo Sap), 12g of Fuling (Poria), 12g of Dangshen (Codonopsis), 15g of Shichun (Ulva), 15g of Haizao (Sargassum), 6g of Shanzha (Crataegus pinnatifida), and 6g of Chaihu (Bupleurum chinense). These were soaked in 8 times the amount of water for 30 minutes, decocted for 30 minutes, filtered, and then decocted again in 8 times the amount of water for 30 minutes. The filtrates were combined and mixed, then concentrated under reduced pressure at 40-60℃ until the concentration was 2g of raw material per ml, suitable for animal experiments. The final dose was 10.14g / kg / day administered via gavage.

[0067] Group 5: High-dose treatment group of Tianfushen Decoction. The patient was fed a high-fat, high-fructose, and high-cholesterol diet. The following herbs were used: 15g of raw Tianzhuhuang (Bamboo Sap), 15g of Poria cocos, 15g of Codonopsis pilosula, 20g of Ulva lactuca, 20g of Sargassum fusiforme, 9g of Crataegus pinnatifida, and 9g of Bupleurum chinense. These were soaked in 10 times the amount of water for 20 minutes, decocted for 40 minutes, filtered, and then decocted again in 10 times the amount of water for 40 minutes. The filtrates were combined and mixed, then concentrated under reduced pressure at 40-60℃ until a concentration of 4g of raw material per ml was reached, suitable for animal experiments. The final dose was 20.28g / kg / day administered via gavage.

[0068] Group 6: Treatment group where Gentian was replaced with Bambusa textilis (Tianzhuhuang). This group was fed a high-fat, high-fructose, and high-cholesterol diet. Gentian, which has similar heat-clearing, phlegm-resolving, liver-clearing, and dampness-removing effects to Bambusa textilis, was used instead of Bambusa textilis. The treatment consisted of 15g of raw Gentian, 15g of Poria cocos, 15g of Codonopsis pilosula, 20g of Ulva prostrata, 20g of Sargassum, 9g of Crataegus pinnatifida, and 9g of Bupleurum chinense. These were soaked in 10 times the amount of water for 20 minutes, decocted for 40 minutes, filtered, and then decocted again in 10 times the amount of water for 40 minutes. The filtrates were then combined and mixed, and concentrated under reduced pressure at 40-60℃ until a concentration of 4g of raw material per ml was reached, suitable for animal experiments. The final treatment was administered at 20.28g / kg / day via gavage.

[0069] Group 7: The group treated with the compound formula without Tianzhuhuang was fed a high-fat, high-fructose, and high-cholesterol diet. Tianzhuhuang, the principal ingredient, was removed. The formula consisted of 15g of raw Poria cocos, 15g of Codonopsis pilosula, 20g of Ulva prostrata, 20g of Sargassum fusiforme, 9g of Crataegus pinnatifida, and 9g of Bupleurum chinense. These were soaked in 10 times the amount of water for 20 minutes, decocted for 40 minutes, filtered, and then decocted again in 10 times the amount of water for 40 minutes. The filtrates were then combined and mixed, and concentrated under reduced pressure at 40-60℃ until a concentration of 4g of raw material per ml was reached, suitable for animal experiments. The final treatment was administered via gavage at a dose of 17.16g / kg / day.

[0070] Group 8: The treatment group without Poria cocos and Codonopsis pilosula was fed a high-fat, high-fructose, and high-cholesterol diet. The Poria cocos and Codonopsis pilosula were removed. The treatment regimen for this group consisted of 15g of raw Bambusa textilis, 20g of Ulva prostrata, 20g of Sargassum fusiforme, 9g of Crataegus pinnatifida, and 9g of Bupleurum chinense. These were soaked in 10 times the amount of water for 20 minutes, decocted for 40 minutes, filtered, and then decocted again in 10 times the amount of water to the residue. The residue was then filtered again. The two filtrates were combined and mixed, and concentrated under reduced pressure at 40-60℃ until the concentration was 4g of raw material per ml, suitable for animal experiments. The final treatment was administered via gavage at a concentration of 14.04g / kg / day.

[0071] Group 9: The group treated with a compound formula excluding Ulva prostrata and Sargassum was fed a high-fat, high-fructose, and high-cholesterol diet. The formula for this group consisted of 15g of raw herbs (Bamboo Sapium sebiferum, Poria cocos, Codonopsis pilosula, Crataegus pinnatifida, and Bupleurum chinense), soaked in 10 times the amount of water for 20 minutes, decocted for 40 minutes, filtered, and then decocted again in 10 times the amount of water for 40 minutes. The filtrates were then combined and mixed, and concentrated under reduced pressure at 40-60℃ until a concentration of 4g of raw material per ml was reached, suitable for animal experiments. The final treatment regimen was 9.88g / kg / day via gavage.

[0072] Group 10: Positive control group, fed a high-fat, high-fructose, and high-cholesterol diet, and treated with polyene phosphatidylcholine 178 mg / kg / d (human equivalent dose) by gavage.

[0073] Detection indicators: The mental state of mice was observed regularly, and their weight was recorded weekly. At the end of the experiment, the liver weight of mice was measured. Serum biochemical analysis was performed to detect blood lipids, liver function, and blood glucose levels, including total cholesterol (TC), triglycerides (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and blood glucose (FBG). Liver tissue pathological staining was used to observe liver inflammation and steatosis in mice.

[0074] Pharmacological studies: Based on previous studies, the intensity of reactive oxygen species (ROS) in mouse liver tissue was detected.

[0075] Experimental methods:

[0076] TG, TC, ALT, AST, and FBG were detected using a fully automated biochemical analyzer.

[0077] The ROS detection method is as follows: Fresh mouse liver tissue was taken, washed with PBS, and 450 μL of homogenate was added to 50 mg of tissue. The tissue was homogenized thoroughly. The mixture was centrifuged at 10000 g at 4℃ for 10 min, the precipitate was discarded, and the supernatant was collected. 90 μL of homogenate buffer, 10 μL of tissue homogenate supernatant, and 1 μL of 100×DHE staining solution were added to a black 96-well plate. The plate was incubated at 37℃ in the dark for 20 min. The absorbance was measured using a microplate reader with an excitation wavelength of 520 nm and an emission wavelength of 605 nm.

[0078] The HE staining method is as follows: Liver tissue samples were removed from formalin fixative and continuously rinsed with running water for 4 hours to remove residual fixative and impurities. The tissue samples were then placed in an automated dehydrator for dehydration. After dehydration, the samples were embedded in paraffin to form paraffin blocks. The paraffin blocks were cut into 5 μm thick sections using a microtome and attached to glass slides. The dried sections were then immersed in xylene for 10 min, repeated once, to completely remove the paraffin. Next, they were immersed in anhydrous ethanol for 5 min, then again for 2 min, followed by immersion in 95%, 90%, 80%, and 70% ethanol for 2 min each. Finally, the sections were rinsed with distilled water and then washed three times with PBS solution for 5 min each time to ensure adequate tissue hydration and removal of impurities. The sections were then immersed in hematoxylin staining solution for 1 min to stain the cell nuclei. The sections were rinsed with distilled water to remove excess staining solution. Next, the sections were immersed in 0.4% hydrochloric acid ethanol for 4 seconds for differentiation, rinsed again with distilled water, and then immersed in 0.6% ammonia water for bluing to make the cell nuclei more clearly stained. Finally, the sections were rinsed with running water. The sections were then stained with eosin for 2 minutes to stain the cytoplasm. The stained sections were then immersed in 95% ethanol for 5 minutes, then in anhydrous ethanol for 5 minutes, and finally in xylene for 5 minutes to completely dehydrate the sections. After the sections were air-dried, they were mounted with neutral resin and photographed.

[0079] The Oil Red O staining method is as follows: Pre-cool the cryostat to ensure the machine temperature is stable at a suitable low temperature. Remove the frozen tissue sample and, after the sample temperature has equilibrated, thaw it on ice in preparation for subsequent sectioning. Attach the thawed tissue sample to the cryostat holder using OCT adhesive, ensuring the sample is firmly fixed. Place the specimen holder with the attached sample on the pre-cooled cryostat and, after the OCT adhesive has solidified, section it to a thickness of 8 μm. Gently attach the section to a pre-labeled glass slide, ensuring the section adheres firmly. Remove the frozen section and rinse it in a container of 60% isopropanol for 20 seconds to remove impurities and residual OCT adhesive from the section surface. Mix Oil Red O dye and diluent at a ratio of 5:2 and filter twice with filter paper to ensure the staining solution is homogeneous and free of impurities. Immerse the section in the Oil Red O staining solution for 10 minutes to ensure the lipid droplets are fully stained. After staining, rinse the sections with 60% isopropanol for 20 seconds to remove excess Oil Red O staining solution, then wash the sections with preheated double-distilled water at 37°C to ensure the section surface is clean. Counterstain the sections with hematoxylin (counterstain) for 4 minutes to stain the cell nuclei. Mount the sections with a water-based mounting medium preheated to liquid state at 60°C and photograph them.

[0080] Statistical analysis methods:

[0081] Data were processed using GraphPadPrism 7.0 software, and are expressed as mean ± standard deviation. One-way ANOVA was used for comparisons between groups, and repeated measures ANOVA was used for comparisons of repeated measures data. P < 0.05 was considered statistically significant.

[0082] Performance results:

[0083] Tianfu Shentang can reduce HFD-induced increases in body weight and liver weight in mice, lower levels of TG, TC, ALT, AST, and FBG, and improve blood lipids, liver function, blood glucose, as well as hepatic steatosis and inflammatory response. Its mechanism of action is related to oxidative stress. Details are as follows:

[0084] Figure 1This study investigated the effect of Tianfushen Decoction intervention on the body weight of HFD-induced MASLD mice. During the modeling period, the mice were in good spirits, agile, and had normal eating and drinking habits. At the beginning of the experiment, there was no significant difference in body weight among the groups (P > 0.05). During the modeling treatment, mice fed a high-fat, high-fructose, and high-cholesterol diet showed rapid body weight gain, with the model group (group 2) showing the fastest growth. By the end of the experiment, the body weight of the normal group (group 1) remained relatively stable, while the body weight of the model group (group 2) continued to increase, showing a statistically significant difference compared to the normal group (group 1) (P < 0.05). The body weight of mice in each treatment group also continued to increase, but the rate of increase was significantly lower than that in the model group (group 2). At the end of the experiment, compared to the model group (group 2), the medium- and high-dose treatment groups (groups 4 and 5) were significantly lighter, with statistically significant differences (P < 0.05). Replacing *Bamboo Sap* with *Gentiana scabra* (Group 6), omitting *Bamboo Sap* (Group 7), omitting the *Codonopsis pilosula* and *Poria cocos* pair (Group 8), omitting the *Ulva lactuca* and *Sargassum fusiforme* pair (Group 9), or the positive control (Group 10) all resulted in a decrease in body weight compared to the model group (Group 2), but the differences were not statistically significant (P>0.05). This indicates that the combination of all the herbs in the Tianfu Shen Tang formula has the best intervention effect on mouse body weight.

[0085] Figure 2 This study investigated the effect of Tianfushen Decoction intervention on liver weight in HFD-induced MASLD mice. Compared with the normal group (group 1), the liver weight in the model group (group 2) was significantly increased (P < 0.05); while compared with the model group (group 2), the liver weight in each treatment group (groups 3, 4, and 5) was significantly decreased, with statistically significant differences (P < 0.05). Compared with the model group (group 2), the liver weight of mice with Gentiana scabra replacing Bambusa textilis (group 6) was also significantly decreased, with statistically significant differences (P < 0.05); while compared with the high-dose treatment group (group 5), the liver weight of mice without Bambusa textilis (group 7) was significantly increased, with statistically significant differences (P < 0.05). The liver-to-body weight ratio was calculated, and the results were similar to those for liver weight. Among them, the liver weight of the positive control group (group 10) was significantly increased compared with the high-dose treatment group (group 5), with statistically significant differences (P < 0.05). This indicates that the entire formula of Tianfushen Decoction has the best intervention effect on liver weight in mice, and its efficacy is superior to that of the positive control.

[0086] Figure 3This study investigated the effects of Tianfushen Decoction intervention on blood lipids in HFD-induced MASLD mice. Compared with the normal group (group 1), the serum TG content in the model group (group 2) was significantly increased (P < 0.05); while compared with the model group (group 2), the serum TG content in the medium- and high-dose treatment groups (groups 4 and 5) was significantly decreased, with statistically significant differences (P < 0.05). Compared with the model group (group 2), the serum TG content in mice with Gentiana scabra replacing Bambusa textilis (group 6) was also significantly decreased, with statistically significant differences (P < 0.05); while compared with the high-dose treatment group (group 5), the serum TG content in mice without Bambusa textilis (group 7) was significantly increased, with statistically significant differences (P < 0.05). Compared with the normal group (group 1), the serum TC content in the model group (group 2) was significantly increased (P < 0.05); while compared with the model group (group 2), the serum TC content in the high-dose treatment group (group 5) was significantly decreased, with statistically significant differences (P < 0.05). Replacing *Bamboo Sap* with *Gentiana scabra* (Group 6), omitting *Bamboo Sap* (Group 7), omitting the *Codonopsis pilosula* and *Poria cocos* pair (Group 8), omitting the *Ulva lactuca* and *Sargassum fusiforme* pair (Group 9), or using the positive control (Group 10) all resulted in a decrease in serum TG levels compared to the model group (Group 2), but the differences were not statistically significant (P > 0.05). This indicates that the entire formula of Tianfu Shen Tang has the best intervention effect on blood lipids in mice.

[0087] Figure 4 This study investigated the effect of Tianfushen Decoction intervention on blood lipids in HFD-induced MASLD mice. Compared with the normal group (group 1), the serum ALT and AST levels in the model group (group 2) were significantly increased (P < 0.05); while compared with the model group (group 2), the serum ALT and AST levels in the medium- and high-dose treatment groups (groups 4 and 5) were significantly decreased, with statistically significant differences (P < 0.05). This indicates that Tianfushen Decoction can improve liver function in MASLD mice.

[0088] Figure 5 This study investigated the effect of Tianfushen Decoction intervention on blood glucose levels in HFD-induced MASLD mice. Compared with the normal group (group 1), the serum FBG level in the model group (group 2) was significantly increased (P < 0.05); while compared with the model group (group 2), the serum FBG level in the medium- and high-dose treatment groups (groups 4 and 5) was significantly decreased, with statistically significant differences (P < 0.05). This indicates that Tianfushen Decoction can improve blood glucose levels in MASLD mice.

[0089] Figure 6This study investigated the effects of Tianfushen Decoction on liver histopathology in HFD-induced MASLD mice. Compared with the normal group (group 1), the model group (group 2) showed significant steatosis, fatty vacuoles, and inflammatory cell infiltration in the liver tissue. Compared with the model group (group 2), liver inflammation and steatosis were improved in all treatment groups. The most significant improvement was observed in the high-dose treatment group (group 5).

[0090] Figure 7 This study investigated the effect of Tianfushen Decoction on oxidative stress levels in the liver tissue of HFD-induced MASLD mice. Compared with the normal group (group 1), the ROS level in the liver of mice in the model group (group 2) was significantly increased (P < 0.05); while compared with the model group (group 2), the ROS level in the liver of mice in the medium- and high-dose treatment groups (groups 4 and 5) was significantly decreased, with statistically significant differences (P < 0.05). Compared with the high-dose treatment group (group 5), the ROS level in the liver of mice treated with the *Ulva lactuca* and seaweed combination (group 9) and the positive control (group 10) was significantly increased, with statistically significant differences (P < 0.05). This indicates that the entire formula of Tianfushen Decoction has the best intervention effect on liver weight in mice, and its efficacy is superior to that of the positive control.

[0091] Clinical efficacy:

[0092] In clinical practice, Tianfu Shentang is used for patients who clearly meet the diagnostic criteria for MASLD and also meet the diagnostic criteria for phlegm-dampness-blood stasis-heat syndrome in traditional Chinese medicine. Specifically, the diagnostic criteria for MASLD are: histological, imaging, and blood biomarker evidence of hepatic steatosis, combined with overweight / obesity (BMI > 23 kg / m² in Asian populations). 2 Individuals meeting one of the following three criteria: type 2 diabetes, metabolic dysfunction (with at least two metabolic risk factors). The diagnostic criteria for phlegm-dampness-blood stasis-heat syndrome are: frequent abdominal distension, sometimes accompanied by distending pain and discomfort in the chest and rib area, or accompanied by bitter taste in the mouth, dry mouth, sticky and unsatisfactory stools, or alternating constipation and diarrhea, easy fatigue, heaviness in the body, irritability, and poor mood; the tongue coating is mostly yellow and greasy or white and greasy, and the tongue body is reddish.

[0093] This study included 23 patients diagnosed with MASLD (Hypertension, Dampness, Blood Stasis, and Heat) this year and treated with Tianfu Shentang (a traditional Chinese medicine formula) for 8-12 weeks. The results are shown in Table 1. Statistical analysis was performed using GraphPadPrism 7.0 software. Data conforming to a normal distribution were expressed as mean (standard deviation), while data not conforming to a normal distribution were expressed as median (interquartile range). Paired t-tests or Wilcoxon Signed-Rank tests were used for comparisons between groups before and after treatment, depending on the data type. A p-value < 0.05 was considered statistically significant.

[0094] Table 1. Statistical Table of Clinical Efficacy

[0095]

[0096]

[0097] Controlled attenuation parameter (CAP) of liver elastography was used to show the degree of steatosis. The results showed that the intervention of Tianfushen Decoction could significantly improve the hepatic steatosis in patients with MASLD phlegm-dampness-blood stasis-heat syndrome (P<0.001).

[0098] Meanwhile, the intervention of Tianfu Shentang can also significantly reduce the weight and BMI index of patients with MASLD phlegm-dampness-blood stasis-heat syndrome (P<0.001).

[0099] Serum lipid test results showed that the intervention of Tianfushen Decoction could significantly reduce the triglyceride (TG) content in patients with MASLD phlegm-dampness-blood stasis-heat syndrome (P<0.05).

[0100] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. A Tianfu Shentang composition, characterized in that, The components include the following parts by weight: 9-15 parts of Tianzhuhuang, 9-15 parts of Fuling, 9-15 parts of Dangshen, 10-20 parts of Shichun, 10-20 parts of Haizao, 3-9 parts of Shanzha, and 3-9 parts of Chaihu.

2. The Tianfu Shentang composition according to claim 1, characterized in that, The Tianfushen Decoction composition comprises the following components in parts by weight: 9 parts of Tianzhuhuang, 9 parts of Fuling, 9 parts of Dangshen, 10 parts of Shichun, 10 parts of Haizao, 3 parts of Shanzha, and 3 parts of Chaihu; or, 12 parts of Tianzhuhuang, 12 parts of Fuling, 12 parts of Dangshen, 15 parts of Shichun, 15 parts of Haizao, 6 parts of Shanzha, and 6 parts of Chaihu. or, 15 parts of Tianzhuhuang (Bamboo Sap), 15 parts of Fuling (Poria), 15 parts of Dangshen (Codonopsis), 20 parts of Shichun (Ulva), 20 parts of Haizao (Sargassum), 9 parts of Shanzha (Crataegus pinnatifida), and 9 parts of Chaihu (Bupleurum chinense).

3. The Tianfu Shentang composition according to claim 1, characterized in that, The Tianfushen Decoction composition comprises the following components in parts by weight: 12 parts of Tianzhuhuang (Bamboo Sap), 12 parts of Fuling (Poria), 12 parts of Dangshen (Codonopsis), 15 parts of Shichun (Ulva), 15 parts of Haizao (Sargassum), 6 parts of Shanzha (Crataegus pinnatifida), and 6 parts of Chaihu (Bupleurum chinense).

4. The Tianfu Shentang composition according to claim 1, characterized in that, All components in the Tianfushen Decoction composition are raw medicinal materials.

5. A method for preparing the Tianfu Shentang composition as described in any one of claims 1-4, characterized in that, Includes the following steps: S1: Weigh each component according to the formula, soak in water, boil and filter, and collect the first filtrate. S2: Take the filter residue, add water again and boil, then filter and collect the secondary filtrate; S3: Combine the two filtrates and mix them well to obtain Tianfu Shentang.

6. The method for preparing a Tianfu Shentang composition according to claim 5, characterized in that, Step S1 includes one or more of the following: i) The mass of water added should be 6-10 times the total mass of all components in the formula; ii) Soaking time is between 20 and 40 minutes; iii) The simmering time is between 20 and 40 minutes.

7. The method for preparing a Tianfu Shentang composition according to claim 5, characterized in that, Step S2 includes one or both of the following: i) The mass of water added should be 6-10 times the mass of the filter cake; ii) The simmering time is between 20 and 40 minutes.

8. The method for preparing a Tianfu Shentang composition according to claim 5, characterized in that, It also includes the following steps: S4: The Tianfu Shentang obtained in step S3 is concentrated under reduced pressure to prepare a concentrated Tianfu Shentang solution of a specified concentration.

9. The method for preparing a Tianfu Shentang composition according to claim 8, characterized in that, Step S4 includes one or both of the following: i) The vacuum concentration is carried out at 40-60°C; ii) The concentration of the Tianfushen Decoction concentrate is 1-4g of raw material per milliliter.

10. The use of the Tianfushen Decoction composition as described in any one of claims 1-4 in the preparation of a medicament for metabolic dysfunction-related fatty liver disease.

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