A traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the methods of soothing liver and strengthening spleen, promoting blood circulation and removing blood stasis

Abstract

The present application relates to the technical field of traditional Chinese medicine compound preparation, in particular to a traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on soothing liver and invigorating spleen, activating blood and resolving stasis, which is characterized by comprising the following raw materials in parts by weight: 6-14 parts of Radix Bupleuri, 8-16 parts of Radix Scutellariae, 10-20 parts of Radix Codonopsis, 10-20 parts of Carapax Trionycis Acetate, 6-14 parts of Eupolyphaga, 6-14 parts of Semen Persicae, and 10-20 parts of Curcuma. The present application can significantly improve the clinical curative effect of liver fibrosis and cirrhosis, and the clinical total effective rate of primary biliary cholangitis liver fibrosis (cirrhosis) is improved to 90.63%, which has a good application prospect.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine compound preparation technology, specifically to a traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the methods of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis. Background Technology

[0002] Liver fibrosis is an inevitable pathological process in the progression of various chronic liver injuries to cirrhosis. Its essence lies in the imbalance between excessive deposition and degradation of extracellular matrix within the liver. The course of liver fibrosis is characterized by its insidious nature and chronic protracted progression. Patients without standardized treatment will sequentially experience the cholangitis stage, the progressive fibrosis stage, and the cirrhosis stage during this long chronic course, eventually progressing to decompensated cirrhosis and developing serious complications such as esophageal and gastric variceal bleeding and hepatic encephalopathy. At the pathophysiological level, this process involves the cascade release of inflammatory factors, abnormal activation of hepatic stellate cells, and overall dysbiosis of the gut microbiota. Although modern medicine has made progress in treating the underlying causes, there is currently a lack of internationally recognized effective drugs for prevention and treatment that directly target fibrotic lesions, promote the degradation of deposited matrix, and systematically repair the gut-hepatic axis.

[0003] Current Western medicine treatments mainly target single pathological aspects of liver fibrosis and cirrhosis, making it difficult to simultaneously cover multiple mechanisms such as inflammatory response, fibrosis formation, and gut-liver axis imbalance. This results in limited efficacy and an inability to effectively reverse the fibrosis process. Although the role of gut microbiota in liver fibrosis has received attention, there is a lack of systematic experimental evidence based on compound interventions, especially a lack of comprehensive research on the synergistic regulation of the intestinal barrier, microbiota structure, and metabolic pathways.

[0004] Researchers have been developing traditional Chinese medicine formulas for the treatment of liver fibrosis and cirrhosis, and these formulas have been used in clinical practice to some extent. However, most of the existing traditional Chinese medicine formulas focus on a single target and have low biochemical response rates, insufficient anti-fibrotic effects, and cannot effectively address both symptom relief and pathological reversal. Developing formulas that achieve synergistic effects from multiple targets and produce a significant overall effect has always been a challenge and a key focus of research and development. Summary of the Invention

[0005] The technical problem solved by this invention is to provide a traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the methods of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis. This solves the problems of existing technologies, such as single drug targets, low biochemical response rate, insufficient anti-fibrotic effect, and inability to effectively address both symptom relief and pathological reversal.

[0006] The technical problem solved by this invention is achieved by the following technical solution: A traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis, is composed of the following raw materials in parts by weight: Bupleurum chinense 6-14 parts, Scutellaria baicalensis 8-16 parts, Codonopsis pilosula 10-20 parts, vinegar-processed turtle shell 10-20 parts, Eupolyphaga sinensis 6-14 parts, Prunus persica 6-14 parts, and Curcuma zedoaria 10-20 parts.

[0007] Furthermore, it is composed of the following raw materials in parts by weight: Bupleurum chinense 8-12 parts, Scutellaria baicalensis 10-15 parts, Codonopsis pilosula 12-18 parts, vinegar-processed turtle shell 12-18 parts, Eupolyphaga sinensis 8-12 parts, Prunus persica 8-12 parts, and Curcuma zedoaria 12-18 parts.

[0008] Furthermore, it is composed of the following raw materials in parts by weight: Bupleurum chinense 9-11 parts, Scutellaria baicalensis 11-14 parts, Codonopsis pilosula 14-16 parts, vinegar-processed turtle shell 14-16 parts, Eupolyphaga sinensis 9-11 parts, Prunus persica 9-11 parts, and Curcuma zedoaria 14-16 parts.

[0009] Furthermore, it is composed of the following ingredients in parts by weight: 10 parts Bupleurum, 12 parts Scutellaria baicalensis, 15 parts Codonopsis pilosula, 15 parts vinegar-processed turtle shell, 10 parts Eupolyphaga sinensis, 10 parts peach kernel, and 15 parts Curcuma zedoaria.

[0010] Furthermore, the preparation is a decoction or powder.

[0011] Furthermore, the preparation method of traditional Chinese medicine compound preparations includes the following steps: (1) Weigh each raw material according to the proportion, add water at a ratio of 1:5~10 and soak for 50~70 minutes. After soaking, decoct for 20~40 minutes. After decoction, filter and take the first decoction. (2) Add water to the dregs after the first decoction at a ratio of 1:5 to 10 and decoct for 10 to 30 minutes. After decoction, filter to obtain the second decoction. (3) Combine the first and second medicinal solutions, then concentrate and fill them into a decoction, or dry them into a powder.

[0012] Furthermore, the powder is a freeze-dried powder produced by vacuum freeze-drying, with each 1 gram of freeze-dried powder equivalent to 4 grams of raw medicinal material.

[0013] Furthermore, the concentration of the decoction is 1.5~2.0 g / mL of raw medicinal materials.

[0014] The development of liver fibrosis and cirrhosis involves a complex process of the circulation of Qi, blood, and body fluids throughout the body. Its core pathogenesis is liver stagnation and spleen deficiency, along with Qi stagnation and blood stasis. The liver governs the free flow of Qi, while the spleen governs transformation and transportation, serving as the source of Qi and blood. Prolonged emotional disharmony or pathogenic invasion can lead to liver Qi stagnation and excessive Wood element overacting on Earth element, resulting in impaired spleen function and insufficient production of Qi and blood. Prolonged Qi stagnation inevitably leads to sluggish blood flow and obstruction of liver collaterals, ultimately causing malnourishment of the liver and stagnation of collaterals, gradually forming liver fibrosis and cirrhosis. This pathological state is not limited to the liver but also affects systemic metabolism through the gut and liver axis. The formula described in this invention is based on a treatment strategy of soothing the liver and strengthening the spleen, promoting blood circulation and removing blood stasis. Through the synergistic effect of multiple components and multiple targets, it not only directly acts on the target points of liver fibrosis but also regulates the structure and metabolic homeostasis of the intestinal flora, such as increasing beneficial bacteria like Lactobacillus and Bifidobacterium, and improving bile acid metabolism, thereby blocking disease progression at the systemic level.

[0015] The development of liver fibrosis and cirrhosis involves a complex process of the circulation of Qi, blood, and body fluids throughout the body. Its core pathogenesis is liver stagnation and spleen deficiency, along with Qi stagnation and blood stasis. The liver governs the free flow of Qi, while the spleen governs transformation and transportation, serving as the source of Qi and blood. Prolonged emotional disharmony or pathogenic invasion can lead to liver Qi stagnation and excessive Wood overacting on Earth, resulting in impaired spleen function and insufficient production of Qi and blood. Prolonged Qi stagnation inevitably leads to sluggish blood flow and obstruction of liver collaterals, ultimately causing liver malnourishment and blood stasis, gradually forming liver fibrosis and cirrhosis. This pathological state is not limited to the liver but also affects systemic metabolism through the gut-liver axis. The formula described in this invention is based on a treatment strategy of soothing the liver and strengthening the spleen, promoting blood circulation and removing blood stasis. Through the synergistic effect of multiple components and multiple targets, it not only directly targets liver fibrosis but also regulates the structure and metabolic homeostasis of the intestinal flora, such as increasing beneficial bacteria like Lactobacillus and Bifidobacterium, and improving bile acid metabolism, thereby blocking disease progression at the systemic level.

[0016] In terms of herbal compatibility, this invention uses Bupleurum and vinegar-processed turtle shell as the principal herbs. Bupleurum is bitter and slightly cold in nature, and it specifically enters the liver and gallbladder meridians. It is good at soothing the liver and relieving depression, regulating the flow of Qi, opening up stagnation in the liver meridian, and restoring the liver's normal function of regulating Qi. It is an essential herb for regulating liver Qi. Vinegar-processed turtle shell is salty and cold in nature, and it enters the liver, spleen, and kidney meridians. It is good at softening hard masses and dispersing nodules, and nourishing liver Yin. It can penetrate deep into the liver collaterals to search for long-standing and stubborn stagnation and dissipate masses under the ribs. It can also protect the liver's Yin and blood, and remove blood stasis without damaging Yin fluids. It is an excellent product for treating liver fibrosis and accumulation. The combination of the two herbs, one soothes Qi and the other disperses nodules, one regulates the liver and the other nourishes the liver, directly targeting the core pathogenesis of liver Qi stagnation and blood stasis in the liver collaterals. Scutellaria baicalensis, Curcuma zedoaria, and Eupolyphaga sinensis are used as assistant herbs. Scutellaria baicalensis is bitter and cold, clearing heat and purging, and is good at clearing heat from the liver and gallbladder. When combined with Bupleurum chinense, it clears and purifies simultaneously, dispersing stagnation of Qi and removing accumulated heat, thus relieving the harm of scorching heat in the collaterals and bile leakage. Curcuma zedoaria is bitter, pungent, warm, and invigorating, and is a blood-activating herb. It can both break up Qi to relieve liver stagnation and activate blood to disperse blood stasis, thus promoting the smooth flow of Qi and blood and eliminating masses. Eupolyphaga sinensis is salty and cold, entering the collaterals, and is good at breaking up blood stasis and eliminating latent pathogens. When used with vinegar-processed turtle shell, it greatly enhances the ability to soften hardness, unblock collaterals, disperse blood stasis, and eliminate accumulations. The three herbs assist the principal herb, working together to clear heat, soothe the liver, break up blood stasis, and soften hardness. Peach kernel and codonopsis are used as adjuvant herbs. Peach kernel is bitter and sweet in nature, and neutral in nature. It invigorates blood, removes blood stasis, moistens dryness, and harmonizes blood. It assists the adjuvant herbs in promoting blood circulation, dispersing blood stasis, and relieving pain. Its nourishing nature can counteract the pungent and drying properties of other blood-breaking herbs. Codonopsis is sweet and warm in nature. It invigorates the spleen, replenishes qi, and nourishes the middle jiao. Following the principle that liver disease affects the spleen, it is necessary to strengthen the spleen first. It supports the spleen earth to resist the overacting liver wood, replenishes the source of qi and blood, and supports the body's resistance to expel pathogens. This ensures that the attack does not harm the body's vital energy, and strengthens the spleen to eliminate the source of internal dampness and turbidity. The seven herbs in the whole formula are well-matched, with the principal and adjuvant herbs in a proper order. All the herbs enter the liver and spleen meridians and directly reach the disease site, without the need for additional guiding herbs. In terms of the principles of the prescription, liver fibrosis falls under the categories of hypochondriac pain and abdominal masses in Traditional Chinese Medicine (TCM). Its underlying cause is liver stagnation and spleen deficiency, leading to overall organ weakness. The symptoms include qi stagnation and blood stasis, with dampness, turbidity, and heat accumulating in the liver meridians. With prolonged illness, the disease penetrates the meridians, causing qi and blood to stagnate and gradually form masses. This TCM compound preparation utilizes the principles of soothing the liver and strengthening the spleen, regulating qi and promoting blood circulation, softening the liver and dispersing masses. Soothing the liver and regulating qi is the primary focus, ensuring smooth qi flow so that bile flows normally and turbidity cannot accumulate. Strengthening the spleen and supporting the body's vital energy is the foundation, ensuring healthy function in the middle jiao (spleen and stomach), resulting in abundant qi and blood and strong vital energy, leaving no room for pathogenic factors to invade. Promoting blood circulation and softening masses is the key, clearing stasis, unblocking meridians, and gradually eliminating masses. This formula harmonizes the liver and spleen, treats both qi and blood, and addresses both deficiency and excess. It soothes the liver without depleting liver yin, strengthens the spleen without stagnation of qi, removes blood stasis without harming the body's vital energy, and softens hardened masses without damaging the spleen and stomach. It deeply understands the pathogenesis of this disease, which is characterized by deficiency in the root and excess in the branches, and a mixture of deficiency and excess. It can soothe liver stagnation, replenish spleen deficiency, clear blood stasis, and harmonize liver meridians, thereby reversing liver fibrosis, delaying disease progression, and improving various clinical symptoms.

[0017] Compared with the traditional Chinese medicine compound turtle shell liver-softening tablets, this compound Chinese medicine preparation has significant differences in its pathogenesis approach, treatment focus, compatibility characteristics, and applicable stages. The traditional compound turtle shell liver-softening tablets take softening and dispersing masses, resolving blood stasis and detoxifying, and tonifying qi and nourishing blood as its core treatment principles. The formula is based on blood-nourishing ingredients such as turtle shell, cordyceps, and placenta, which focus on nourishing the liver and kidneys and dispersing blood stasis and eliminating accumulations. It mainly targets the pathogenesis of liver cirrhosis with liver and kidney deficiency and accumulation of blood stasis and toxins. The overall medicinal properties are insufficient in soothing the liver and regulating qi and clearing gallbladder heat. For the early and middle stages of primary biliary cholangitis with liver fibrosis, patients often have symptoms of liver and gallbladder stagnation and lingering damp heat. Using only nourishing and softening ingredients can easily obstruct the flow of qi and trap pathogens, which may aggravate discomfort such as hypochondriac distension and epigastric fullness. Moreover, it is not very targeted at the core aspects of this disease, namely liver and gallbladder dysfunction and bile stasis. The innovation of this invention lies first in its deepened understanding of the pathogenesis. It closely adheres to the core pathogenesis of primary biliary cholangitis, specifically liver stagnation and spleen deficiency, qi stagnation and blood stasis. It addresses both the superficial symptoms of liver meridian obstruction and the underlying causes of impaired liver and gallbladder function and spleen dysfunction, thus regulating qi and blood simultaneously. Secondly, it employs precise treatment combinations, integrating methods for soothing the liver, clearing the gallbladder, strengthening the spleen, promoting blood circulation, and softening hardness into a single formula. The combination of Bupleurum and Scutellaria baicalensis directly targets the core pathological links of bile stasis and liver and gallbladder stagnation, promoting bile excretion and improving liver function. It effectively addresses cholestasis, a characteristic not found in Compound Turtle Shell Softening Liver Tablets. Furthermore, the formula balances tonification and purgation, focusing on regulating and purging while simultaneously purging, ensuring neither stagnation nor harm. This approach better aligns with the progression of primary biliary cholangitis from chronic cholangitis to liver fibrosis. For early and mid-stage patients, it promotes blood circulation and softens hardened masses while clearing heat and stagnant qi, avoiding the drawbacks of cloying and cloying substances that obstruct qi flow and exacerbate dampness. Finally, it combines disease and syndrome differentiation. This invention adheres to the principles of TCM syndrome differentiation and treatment while closely considering the characteristics of liver fibrosis, achieving a unity of disease and syndrome differentiation, thus better suited to clinical practice.

[0018] Beneficial Effects: The traditional Chinese medicine compound preparation of this invention, based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis, for the treatment of liver fibrosis and cirrhosis, firstly significantly improves the clinical efficacy of liver fibrosis and cirrhosis. Treatment with this traditional Chinese medicine compound preparation increases the overall clinical effective rate of primary biliary cholangitis-related liver fibrosis (cirrhosis) to 90.63%, particularly enhancing the regulation of the core cholestasis indicators ALP and GGT, thus overcoming the clinical deficiency of insufficient response to Western medicine alone. Secondly, the traditional Chinese medicine compound preparation of this invention can precisely reverse the pathological process of liver fibrosis and cirrhosis. Imaging and serological indicators confirm that this compound preparation can significantly reduce liver stiffness, decrease extracellular matrix deposition, and greatly optimize the staging of liver fibrosis, providing empirical evidence for the reversal of cirrhosis. Furthermore, the preparation of this invention can regulate the gut-hepatic axis microenvironment, achieving holistic treatment of chronic liver disease from a systems biology perspective by reshaping gut microbiota diversity, inhibiting the spread of pathogenic bacteria, repairing the integrity of the intestinal barrier, and clearing enterogenic circulating toxins. Furthermore, the preparation process of this invention is simple and safe, the compound preparation has a rigorous formulation and batch stability, and there are no serious adverse reactions in clinical application, thus showing good application prospects. Attached Figure Description

[0019] Figure 1 The results of diversity indicators for the treatment group and the control group were statistically analyzed.

[0020] Figure 2 This is a paired statistical analysis of the relative abundance of beneficial bacteria genera in the treatment and control groups.

[0021] Figure 3 This section presents statistical data on individual paired changes in target metabolites between the treatment and control groups. Detailed Implementation

[0022] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below with reference to specific embodiments.

[0023] Example 1

[0024] The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the method of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis described in this embodiment is composed of the following raw materials in parts by weight: Bupleurum chinense 10 parts, Scutellaria baicalensis 12 parts, Codonopsis pilosula 15 parts, vinegar-processed turtle shell 15 parts, Eupolyphaga sinensis 10 parts, Prunus persica 10 parts, and Curcuma zedoaria 15 parts.

[0025] The preparation process includes the following steps: (1) Weigh each raw material according to the proportion, add water at a ratio of 1:8 and soak for 60 minutes. After soaking, decoct for 30 minutes. After decoction, filter and take the first decoction. (2) Add water to the dregs after the first decoction at a ratio of 1:6 and decoct for 20 minutes. After decoction, filter to obtain the second decoction. (3) Combine the first and second medicinal solutions, then concentrate and fill them into 100mL / bag decoctions, or use vacuum freeze-drying technology to dry them into powder.

[0026] Example 2

[0027] The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the method of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis described in this embodiment is composed of the following raw materials in parts by weight: Bupleurum chinense 10 parts, Scutellaria baicalensis 10 parts, Codonopsis pilosula 12 parts, vinegar-processed turtle shell 16 parts, Eupolyphaga sinensis 9 parts, Prunus persica 12 parts, and Curcuma zedoaria 15 parts.

[0028] The preparation method is the same as in Example 1.

[0029] Example 3

[0030] The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the method of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis described in this embodiment is composed of the following raw materials in parts by weight: Bupleurum chinense 12 parts, Scutellaria baicalensis 10 parts, Codonopsis pilosula 13 parts, vinegar-processed turtle shell 12 parts, Eupolyphaga sinensis 12 parts, Prunus persica 8 parts, and Curcuma zedoaria 17 parts.

[0031] The preparation method is the same as in Example 1.

[0032] Compare with Example 1 The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis described in this comparative example is composed of the following raw materials in parts by weight: Bupleurum chinense 20 parts, Scutellaria baicalensis 8 parts, Codonopsis pilosula 8 parts, vinegar-processed turtle shell 10 parts, Eupolyphaga sinensis 6 parts, Prunus persica 20 parts, and Curcuma zedoaria 6 parts.

[0033] The preparation method is the same as in Example 1.

[0034] Compare with Example 2 The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis described in this comparative example is composed of the following raw materials in parts by weight: Bupleurum chinense 10 parts, Scutellaria baicalensis 12 parts, Codonopsis pilosula 15 parts, vinegar-processed turtle shell 10 parts, Carthamus tinctorius 10 parts, Prunus persica 12 parts, and Sparganium stoloniferum 15 parts.

[0035] (a) Identification of chemical composition The chemical components in Example 1 were identified using ultra-high performance liquid chromatography-tandem high-resolution mass spectrometry. Forty-one key chemical components were preliminarily identified, covering flavonoids and their glycosides, triterpenoid saponins, phenolic acids, cyanogenic glycosides, and long-chain fatty acids.

[0036] The chemical composition identification results of the decoction prepared in Example 1 are shown in Table 1.

[0037] Table 1. Identification results of the chemical components of the decoction described in Example 1.

[0038] Network pharmacology analysis revealed that the active ingredients in the compound preparation of this invention target core targets including TP53, TNF, SRC, AKT1, and STAT3. KEGG enrichment analysis showed that they mainly act on the PI3K-Akt signaling pathway, AGE-RAGE signaling pathway, and TNF signaling pathway, and are widely involved in immune regulation and fibrosis reversal.

[0039] (II) Animal Experiment Verification The efficacy of the traditional Chinese medicine compound preparation described in this invention was evaluated using a cholestatic liver fibrosis rat model induced by bile duct ligation (BDL).

[0040] The experiment included a sham surgery group, a model group, a UDCA group, and drug administration groups (control group 1, control group 2, example group 1, example group 2, and example group 3), with 6 rats randomly assigned to each group. According to the established protocol, the model group, UDCA group, and CBD group underwent bile duct separation surgery, while the sham surgery group only underwent bile duct separation without ligation. After surgery, penicillin was injected and administered via abdominal gavage. The rats were fed sterile distilled water and normal feed. After 7 days of observation, the UDCA group was given UDCA (50 mg / kg / day), and the treatment groups were given the corresponding animal-equivalent doses of this traditional Chinese medicine compound preparation (Control Example 1 (6.11 g / kg / day), Control Example 2 (6.58 g / kg / day), Example 1 (6.82 g / kg / day), Example 2 (6.58 g / kg / day), and Example 3 (6.58 g / kg / day). The preparation was a lyophilized powder preparation prepared according to the method described in Example 1 of the specific implementation method, administered continuously for 21 days. At the end of the treatment period, all rats were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (0.15 mL / 100 g). Blood samples were collected via femoral artery puncture, followed by rapid resection of the left lateral lobe of the liver, which was immediately preserved. 80℃ refrigerator.

[0041] The results of the rat serum tests in each group are shown in Table 2.

[0042] Table 2 Statistical table of rat serum test results

[0043] Regarding serum liver function biochemical indicators, compared with the sham-operated group, the serum levels of ALT, AST, ALP, GGT, TBIL, and DBIL in the model group rats were significantly increased, and the ALB level was also significantly upregulated, indicating that the model group rats had obvious hepatocellular damage, cholestasis, and abnormal liver synthetic function, consistent with the typical liver function pathological changes of primary biliary cholangitis and liver fibrosis. Compared with the model group, the UDCA group, control group 1, control group 2, example group 1, example group 2, and example group 3 all downregulated the abnormal expression levels of ALT, AST, ALP, GGT, TBIL, DBIL, and ALB to varying degrees, improving the pathological damage state of liver function. Although control group 1 and control group 2 improved liver function abnormalities to varying degrees, the overall improvement effect was weaker than that of example group 1, and the levels of each indicator were still significantly higher than those in the sham-operated group. The improvement effects of example group 2 and example group 3 were not significantly different from those in example group 1, which is more consistent with the recovery state of normal liver function.

[0044] Overall, the traditional Chinese medicine compound preparations described in this invention can exert the therapeutic effects of protecting the liver, lowering enzymes, promoting bile secretion, and reducing jaundice by reducing hepatocellular damage, improving cholestasis, and correcting abnormal liver synthesis function. The liver function improvement effects of the groups in Examples 2 and 3 are similar to those of the group in Example 1, while the liver function improvement efficacy of the control groups in Examples 1 and 2, which have an unbalanced compatibility ratio, is significantly weakened.

[0045] The results of liver tissue analysis in each group of rats are shown in Table 3.

[0046] Table 3. Statistical table of liver tissue examination results in rats.

[0047] The results showed that, compared with the sham-operated group, the expression levels of core fibrosis markers such as α-smooth muscle actin (α-SMA), hydroxyproline (HYP), matrix metalloproteinase 9 (MMP9), Smad3, and transforming growth factor-β (TGF-β) were significantly increased in the liver tissue of rats in the model group. The expression level of matrix metalloproteinase 2 (MMP-2) was significantly upregulated, while the expression level of tissue inhibitor of metalloproteinases 1 (TIMP-1) was significantly decreased. This suggests that rats in the model group have developed obvious pathological changes of liver fibrosis, and the collagen deposition and fibrosis-related signaling pathways in the liver tissue are in an abnormally activated state. Compared with the model group, the UDCA group, Control Group 1, Control Group 2, Example Group 1, Example Group 2, and Example Group 3 all downregulated the expression levels of α-SMA, HYP, MMP9, Smad3, and TGF-β to varying degrees, and corrected the expression imbalance between MMP-2 and TIMP-1. Among them, Example Group 2 and Example Group 3 showed the most significant improvement effects on the above fibrosis markers. The expression levels of α-SMA, HYP, MMP9, Smad3, and TGF-β were significantly lower than those in Example Group 1, and were closer to the normal physiological levels of the sham-operated group. Although Control Group 1 and Control Group 2 could downregulate the expression of fibrosis markers and improve the MMPs / TIMPs imbalance, the overall improvement effect was weaker than that in Example Group 1. The expression levels of each indicator were still significantly higher than those in the sham-operated group and the UDCA group, showing a gradient difference that was positively correlated with the optimization of the prescription ratio and the anti-liver fibrosis drug efficacy. The expression levels of the indicators in Example Group 2 and Example Group 3 were not much different from those in Example Group 1.

[0048] Overall, the traditional Chinese medicine compound preparation described in this invention can exert a dual therapeutic effect of anti-liver fibrosis and anti-inflammatory liver protection by regulating liver fibrosis-related signaling pathways through multiple targets and inhibiting the excessive release of pro-inflammatory factors. In contrast, the anti-liver fibrosis and anti-inflammatory effects of the control group 1 and control group 2, which had an unbalanced ratio of ingredients, were significantly weakened. This fully demonstrates the scientific and rational nature of the ratio of principal, assistant, adjuvant, and guide herbs in the traditional Chinese medicine compound preparation of this invention, and also reflects the formulation characteristics of traditional Chinese medicine prescriptions that "exquisite combination and proper ratio result in powerful and specific effects".

[0049] (III) Validation of Clinical Results A randomized, parallel-controlled trial was conducted on 68 patients with primary biliary cholangitis and liver fibrosis, with a total intervention period of 24 weeks. The trial was divided into a treatment group (ursodeoxycholic acid combined with the traditional Chinese medicine compound preparation of this invention, n=35) and a control group (ursodeoxycholic acid combined with Compound Turtle Shell Softening Liver Tablets (Inner Mongolia Furui Medical Technology Co., Ltd., National Drug Approval Number Z19991011), n=33). In the treatment group, the traditional Chinese medicine compound preparation of this invention was taken orally as a decoction, twice daily, one sachet (100mL) each time, warm after breakfast and dinner. In the control group, Compound Turtle Shell Softening Liver Tablets were a commercially available drug, a classic traditional Chinese medicine used clinically for anti-liver fibrosis, and should be taken according to the instructions. A total of 61 patients completed the clinical trial (32 in the treatment group and 29 in the control group).

[0050] (1) Imaging and transient liver elastography The liver stiffness score (LSM), abdominal ultrasound score, FIB-4 score, and ARPI score of patients in the treatment group and control group were observed before treatment and at 24 weeks after treatment. The abdominal ultrasound score was determined using the Scheuer criteria for pathological grading of liver fibrosis, as shown in Table 4. The results, as shown in Tables 5 and 6, indicate that after treatment with the traditional Chinese medicine compound preparation described in this invention, the liver ultrasound stage of patients was significantly improved compared to before treatment, the number of cases with normal liver tissue increased, and the number of cases with moderate to severe liver fibrosis significantly decreased. The treatment effect was superior to that of the control group. Furthermore, the LSM of patients using the traditional Chinese medicine compound preparation described in this invention was significantly lower than before treatment, and the reduction in LSM in the treatment group was significantly greater than that in the control group.

[0051] Table 4. Scheuer Ultrasound Staging of the Liver

[0052] Table 5 Comparison of liver ultrasound staging between the two groups before and after treatment ( )

[0053] Table 6 Comparison of instantaneous elasticity index between the two groups before and after treatment ( )

[0054] (2) Detection indicators Observe the changes in liver function indicators (ALT, AST, ALP, GGT, TBIL), blood routine (WBC, RBC, HGb, PLT), and coagulation function (PT, PTA, INR, APTT) before treatment and after 24 weeks of treatment. Also observe the improvement in serum type IV collagen (CGIV), hyaluronic acid (HA), type III procollagen N-terminal peptide (PIIIP), and laminin (LN) indicators of liver fibrosis.

[0055] The results are shown in Tables 7-10. It can be seen that after intervention, serum ALT, AST, ALP, GGT, and TBIL levels in both the treatment and control groups were significantly lower than before treatment. The reduction in ALT, AST, and ALP levels in the treatment group was greater than that in the control group. PLT levels in the treatment group increased after intervention compared to before treatment, and the increase was greater in the treatment group than in the control group. There were no statistically significant differences in PT, PTA, INR, and APTT levels between the two groups before and after treatment. CGIV, HA, PIIIP, and LN levels in the treatment group were all lower than before treatment. The reduction in CGIV, HA, and PIIIP levels in the treatment group was significantly greater than that in the control group. In summary, the traditional Chinese medicine compound preparation described in this invention can protect liver function in patients with liver fibrosis, alleviate liver fibrosis-related indicators, and demonstrate good safety. This indicates that the clinical efficacy of the traditional Chinese medicine compound preparation described in this invention is far superior to that of the currently used classic traditional Chinese medicine compound turtle shell liver-softening tablets, and this invention has better application prospects.

[0056] Table 7 Comparison of liver function indicators between the two groups before and after treatment ( )

[0057] Table 8 Comparison of blood routine indicators between the two groups before and after treatment ( )

[0058] Table 9 Comparison of coagulation function indicators between the two groups before and after treatment ( )

[0059] Table 10 Comparison of liver fibrosis indicators between the two groups before and after treatment ( )

[0060] (3) After the course of treatment, the clinical efficacy was determined with reference to the diagnostic criteria of the "Guidelines for the Diagnosis and Treatment of Primary Biliary Cholangitis (2021)".

[0061] Therapeutic criteria: Complete response (cure): Clinical symptoms and signs disappear or significantly improve; AST, GGT, and ALP levels decrease by ≥50% after treatment; four liver fibrosis indicators improve to the normal range; LSM value recovers to F0-F1 stage.

[0062] Partial response (significant effect): The main clinical manifestations have been partially improved, but satisfactory remission has not yet been achieved. The liver function biochemical indicators AST, GGT, and ALP levels decreased by ≥30% but <50% after treatment, the levels of the four liver fibrosis indicators decreased by ≥30% but <50%, and the LSM value recovered to F1-F2 stage. No response (ineffective): Clinical symptoms do not improve or worsen, and liver function biochemical indicators decrease by <30% in AST, GGT, and ALP levels, and decrease by <30% in the four liver fibrosis markers and LSM value after treatment; Relapse (ineffective): Symptoms reappear after a complete response, accompanied by elevated serum AST, GGT, and ALP levels.

[0063] Response rate = (Number of complete responses + Number of partial responses) / Total number of cases × 100%.

[0064] The results are shown in Table 11. The total effective rate of the treatment group was 90.63%, while that of the control group was 75.86%. The proportion of patients with complete response in the treatment group was significantly higher than that in the control group. Although there was no significant difference in the number of patients with partial response, the number of patients with partial response in the treatment group was greater than that in the control group. The proportion of ineffective cases was significantly lower in the treatment group than in the control group. The difference in the total effective rate between the groups was statistically significant (P<0.05), indicating that the clinical treatment effect of the treatment group was better than that of the control group.

[0065] Table 11 Comparison of overall efficacy between the two groups of patients

[0066] (iv) Validation of gut microbiota remodeling Twenty-four weeks later, fecal and serum samples were collected from 30 patients in the treatment group before and after treatment for bile acid metabolomics and 16S rRNA gut microbiota sequencing. The pre-treatment group was named PRO group, and the post-treatment group was named POST group. In addition, 30 healthy volunteers (CON group) were matched, and their fecal and serum samples were collected for the same bile acid metabolomics and 16S rRNA gut microbiota sequencing to provide a control with the omics changes before and after treatment in the treatment group.

[0067] (1) Comparison of diversity indicators between the treatment group and the control group.

[0068] like Figure 1As shown, the main α-diversity indicators in 16S rRNA gene sequencing are referenced, where af represents Chao1, Observed OTUs, Shannon, Simpson, Pielou evenness, and Faith PD, respectively. Each point represents an independent sample; the gray line connects the pre-treatment and post-treatment measurements of the same treatment subject. It can be seen that the indices such as Chao and Shannon in the post-treatment group are significantly higher than those in the pre-treatment group and are closer to those in the healthy control group, indicating that the traditional Chinese medicine compound preparation described in this invention can significantly improve the α-diversity of the gut microbiota in patients.

[0069] (2) Paired changes in the relative abundance of beneficial bacteria genera in the treatment group and the control group.

[0070] like Figure 2 As shown, AD represents Lactobacillus, Akkermansia, Bifidobacterium, and Subdorianella, respectively. Each point represents an independent sample; the gray line connects the measurements before and after treatment for the same treatment subject. It can be seen that, at the genus level, the traditional Chinese medicine compound preparation described in this invention can significantly upregulate the relative abundance of beneficial bacteria such as Lactobacillus, Akkermansia, and Bifidobacterium.

[0071] (3) Individual pairing changes of target metabolites in the treatment group and the control group.

[0072] like Figure 3 This section presents the statistical results of individual paired changes in the target metabolites in the treatment and control groups, where... 16S rRNA gene sequencing and untargeted metabolomics analysis were performed on fecal and serum samples from patients in the treatment and control groups. In the figure, a represents taurine deoxycholic acid; b represents indoxyl sulfate; c represents hippuric acid; and d represents p-tolyl sulfate. The vertical axis represents log10 (relative signal intensity + 1); statistical tests were based on the unconverted raw signal intensity. Each point represents an independent sample, and gray lines connect measurements before and after treatment for the same patient. q-values ​​were obtained from a two-sided Wilcoxon paired signed-rank test. Metabolomics analysis showed a significant increase in the level of hepatoprotective taurine in the serum of patients in the treatment group, and a significant decrease in the concentrations of enterogenic toxins such as indoxyl sulfuric acid, hippuric acid, and p-tolyl sulfate in the treatment group.

[0073] The above results indicate that the formulation described in this invention can regulate the gut-liver axis microenvironment, and achieve holistic treatment of chronic liver disease from a systems biology perspective by reshaping gut microbiota diversity, inhibiting the spread of pathogenic bacteria, repairing the integrity of the intestinal barrier, and clearing enterogenic circulating toxins.

[0074] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. A traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis, characterized in that... It is composed of the following ingredients in parts by weight: Bupleurum chinense 6-14 parts, Scutellaria baicalensis 8-16 parts, Codonopsis pilosula 10-20 parts, vinegar-processed turtle shell 10-20 parts, Eupolyphaga sinensis 6-14 parts, Prunus persica 6-14 parts, and Curcuma zedoaria 10-20 parts.

2. The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis as described in claim 1, is characterized in that... It is composed of the following ingredients in parts by weight: Bupleurum chinense 8-12 parts, Scutellaria baicalensis 10-15 parts, Codonopsis pilosula 12-18 parts, vinegar-processed turtle shell 12-18 parts, Eupolyphaga sinensis 8-12 parts, Prunus persica 8-12 parts, and Curcuma zedoaria 12-18 parts.

3. The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis as described in claim 2, is characterized in that... It is composed of the following ingredients in parts by weight: Bupleurum chinense 9-11 parts, Scutellaria baicalensis 11-14 parts, Codonopsis pilosula 14-16 parts, vinegar-processed turtle shell 14-16 parts, Eupolyphaga sinensis 9-11 parts, Prunus persica 9-11 parts, and Curcuma zedoaria 14-16 parts.

4. The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis as described in claim 3, is characterized in that... It is composed of the following ingredients in parts by weight: Bupleurum chinense 10 parts, Scutellaria baicalensis 12 parts, Codonopsis pilosula 15 parts, vinegar-processed turtle shell 15 parts, Eupolyphaga sinensis 10 parts, Prunus persica 10 parts, and Curcuma zedoaria 15 parts.

5. The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis as described in claim 1, characterized in that... The preparation is a decoction or powder.

6. The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis as described in claim 1, characterized in that... The preparation method of traditional Chinese medicine compound preparations includes the following steps: (1) Weigh each raw material according to the proportion, add water at a ratio of 1:5~10 and soak for 50~70 minutes. After soaking, decoct for 20~40 minutes. After decoction, filter and take the first decoction. (2) Add water to the dregs after the first decoction at a ratio of 1:5 to 10 and decoct for 10 to 30 minutes. After decoction, filter to obtain the second decoction. (3) Combine the first and second medicinal solutions, then concentrate and fill them into a decoction, or dry them into a powder.

7. The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis as described in claim 6, is characterized in that... The powder is a freeze-dried powder produced by vacuum freeze-drying.

8. The traditional Chinese medicine compound preparation for treating liver fibrosis and cirrhosis based on the principles of soothing the liver and strengthening the spleen, and promoting blood circulation and removing blood stasis as described in claim 6, is characterized in that... The concentration of the decoction is 1.5~2.0 g / m of raw medicinal materials.

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