A traditional Chinese medicine composition for improving liver fibrosis and a preparation method and application thereof

Abstract

This invention provides a traditional Chinese medicine composition for improving liver fibrosis, its preparation method, and its application. The traditional Chinese medicine composition for improving liver fibrosis comprises the following medicinal materials in the indicated weight proportions: 50-70g of *Gynostemma pentaphyllum*, 18-22g of *Artemisia argyi*, and 25-35g of *Bidens pilosa*. The preparation method of the traditional Chinese medicine composition of this invention includes the following steps: taking *Gynostemma pentaphyllum*, *Artemisia argyi*, and *Bidens pilosa*, pulverizing them, adding water and soaking for 20-40 minutes, boiling and extracting for 0.8-1.2 hours after soaking, filtering, adding water to the residue and continuing to boil and extract for 20-40 minutes, collecting the two filtrates, concentrating, and freeze-drying to obtain a dry powder. In this invention's traditional Chinese medicine formula, *Gynostemma pentaphyllum* has the effects of dispersing blood stasis, reducing swelling, and eliminating dampness, and is the principal herb; *Artemisia argyi* warms the middle jiao, invigorates blood circulation, and eliminates dampness, while *Bidens pilosa* clears heat, disperses blood stasis, and reduces swelling. The two herbs work synergistically, using both cold and warm properties to assist *Gynostemma pentaphyllum* in enhancing its effects of dispersing blood stasis and eliminating dampness, and are thus the assistant herbs; the combined use of these herbs has the effect of dispersing blood stasis and eliminating dampness, and has a good effect on treating liver fibrosis, especially on treating liver fibrosis of the dampness-blood stasis type.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine, and in particular to a traditional Chinese medicine composition for improving liver fibrosis, its preparation method, and its application. Background Technology

[0002] Liver fibrosis is a growing global health problem, affecting approximately 1% to 2% of the world's population. The continued progression of liver fibrosis can lead to cirrhosis and liver cancer. Nearly 5% to 30% of cirrhosis patients develop liver cancer within 5 years, and over 80% of liver cancers originate from liver fibrosis or cirrhosis. Current treatment options for liver fibrosis primarily focus on inhibiting the various stages of its formation and progression, including eliminating liver inflammation (silymarin), inhibiting HSC activation (carvedilol), and reducing collagen fiber formation (ferulic acid). Liver transplantation is currently the only potentially curative treatment for end-stage decompensated cirrhosis; however, the severe shortage of donor organs and post-transplant rejection have limited its widespread clinical application. Therefore, how to better alleviate, inhibit, and even reverse liver fibrosis has become a current research hotspot.

[0003] Liver fibrosis is a repair response to the diffuse excessive deposition and abnormal distribution of extracellular matrix (ECM) following chronic liver injury induced by various factors such as viruses, alcohol, autoimmunity, and abnormal metabolism. It is a pathological repair response of the liver to chronic injury, a key step in the progression of various chronic liver diseases to cirrhosis, and an important factor affecting the prognosis of chronic liver diseases. Liver fibrosis is a dynamic process; further development can lead to disordered liver lobule structure, nodular regeneration of hepatocytes, and the formation of pseudolobule structures, i.e., cirrhosis. The core of liver fibrosis development lies in the widespread deposition of ECM within the liver. ECM mainly originates from activated hepatic stellate cells (HSCs). When the liver is subjected to chronic injury or inflammatory stimulation, hepatocytes, Kupffer cells, and other cells release various cytokines that activate stellate cells to secrete large amounts of ECM, forming liver fibrosis and causing liver structural remodeling. The regression of liver fibrosis is often accompanied by a reduction or disappearance of activated HSCs. Therefore, inhibiting HSC activation and inducing activated HSC apoptosis, necrotizing apoptosis, or senescence are effective strategies to improve the pathological progression of liver fibrosis.

[0004] Currently, effective traditional Chinese medicine (TCM) treatments for liver fibrosis often involve complex formulations, high costs, and unsatisfactory efficacy, failing to meet the demand. Therefore, researching TCM formulas for improving liver fibrosis is of great significance. Summary of the Invention

[0005] In view of this, the present invention proposes a traditional Chinese medicine composition for improving liver fibrosis, its preparation method and application. The traditional Chinese medicine formula of the present invention is simple, has low cost, and has good efficacy in treating liver fibrosis, which can better meet the needs.

[0006] The technical solution of this invention is implemented as follows:

[0007] A traditional Chinese medicine composition for improving liver fibrosis is prepared from the following raw materials in the indicated weight ratios: 50-70g of ginger, 18-22g of Artemisia argyi, and 25-35g of Bidens pilosa.

[0008] Furthermore, the traditional Chinese medicine composition is prepared from the following raw materials in the indicated weight proportions: 55-65g of ginger, 18-22g of Artemisia argyi, and 25-35g of Bidens pilosa.

[0009] Furthermore, the traditional Chinese medicine composition is prepared from the following raw materials in the indicated weight proportions: 60g of ginger, 20g of artemisia, and 30g of beggar-ticks.

[0010] Furthermore, the *Gynostemma pentaphyllum* is the dried aerial part of *Gynostemma pentaphyllum*.

[0011] Furthermore, the *Artemisia argyi* is the dried aerial part of *Artemisia argyi*.

[0012] Furthermore, the Bidens pilosa refers to the whole Bidens pilosa plant.

[0013] A traditional Chinese medicine composition for improving liver fibrosis, comprising the traditional Chinese medicine composition described in any one of the present invention.

[0014] The preparation method of the traditional Chinese medicine composition of the present invention includes the following steps: take the herbs of *Gnaphalium affine*, *Artemisia argyi*, and *Bidens pilosa*, crush them, add water and soak for 20-40 minutes, boil and extract for 0.8-1.2 hours after soaking, filter, add water to the filter residue and continue to boil and extract for 20-40 minutes, collect the filtrates twice, concentrate and freeze dry to obtain dry powder.

[0015] The application of any one of the traditional Chinese medicine compositions described in this invention in the preparation of a drug for treating liver fibrosis.

[0016] Furthermore, the application of any of the traditional Chinese medicine compositions described in this invention in the preparation of a medicament for treating liver fibrosis of the dampness-blood stasis type.

[0017] The application of the traditional Chinese medicine composition prepared by the method described in this invention in the preparation of drugs for treating liver fibrosis.

[0018] 1. Costus speciosus, a plant belonging to the genus Costus in the family Zingiberaceae. The dried aerial parts of *Smilax china*. Mainly produced in Hainan, Guangdong, Guangxi, and Yunnan provinces. It has a pungent, sour, and slightly cold taste. It enters the liver, bladder, kidney, and large intestine meridians. It has the functions of dispersing blood stasis and reducing swelling, clearing heat and promoting diuresis. It is used for pertussis, nephritis edema, urinary tract infection, ascites due to liver cirrhosis, and difficulty urinating. Modern research shows that *Smilax china* has effects on the reproductive system (estrogen-like effects, uterine contraction), anti-diabetic, anti-inflammatory, hepatoprotective, and antibacterial effects. The main component of *Smilax china*, diosgenin, has expectorant, desensitizing, anti-inflammatory, lipid-lowering, anti-tumor, and estrogen-like effects.

[0019] 2. *Blumea balsamifera* (L.) DC., a plant belonging to the genus *Blumea* in the family Asteraceae. It is distributed in Hainan, Guangdong, Guangxi, Yunnan, and other regions. It has a pungent and slightly bitter taste; it is warm in nature and enters the lung, liver, and stomach meridians. It possesses the effects of warming the middle jiao and promoting blood circulation, dispelling wind and dampness, and killing parasites. It is mainly used to treat cold-damp diarrhea, abdominal pain and borborygmus, swelling, muscle and bone pain, traumatic injuries, and tinea. Modern research shows that *Blumea balsamifera* has various pharmacological activities such as anti-tumor, analgesic, and anti-inflammatory effects. It can inhibit lipid peroxidation, scavenge free radicals, and has significant antioxidant effects, and it has a protective effect against liver damage.

[0020] 3. Bidens pilosa L., a plant belonging to the genus Bidens in the family Asteraceae. Widely distributed throughout China. It tastes sweet, bland, and bitter, and is slightly cold in nature. It enters the liver, lung, and large intestine meridians. It has the effects of clearing heat and detoxifying, dispersing blood stasis and reducing swelling. It is used for sore throat, diarrhea, dysentery, jaundice, appendicitis, boils, and carbuncles. Modern research shows that Bidens pilosa has anti-inflammatory, antibacterial, hepatoprotective, anti-diabetic, and antimalarial effects. The flavonoid extract of Bidens pilosa has significant therapeutic effects on acute liver injury and liver fibrosis.

[0021] Compared with the prior art, the beneficial effects of the present invention are:

[0022] (1) The traditional Chinese medicine formula of this invention is composed of *Gnaphalium affine*, *Artemisia argyi*, and *Bidens pilosa*. Among them, *Gnaphalium affine* has the effect of dispersing blood stasis, reducing swelling and removing dampness, and is the principal drug; *Artemisia argyi* warms the middle jiao, invigorates blood and removes dampness, and *Bidens pilosa* clears heat, disperses blood stasis and reduces swelling. The two drugs work together, using both cold and warm properties, to help *Gnaphalium affine* to enhance its effect of dispersing blood stasis and removing dampness, and together they are the assistant drugs. The prescription is well-composed and the drugs complement each other. When used together, the drugs have the effect of dispersing blood stasis and removing dampness, and have a good effect on treating liver fibrosis, especially the liver fibrosis of the dampness and blood stasis type.

[0023] (2) The traditional Chinese medicine formula of the present invention can better reduce the levels of HA, IV, III, AST, ALT, ALP, TBIL, DBILI, IBIL and TBA, and has a better effect on improving liver fibrosis and significantly reducing liver damage.

[0024] (3) The traditional Chinese medicine formula of the present invention for treating liver fibrosis is simple, has low cost, and has good efficacy, which can better meet the needs. Attached Figure Description

[0025] Figure 1-4 HE-stained images (10.0x) of liver tissue sections from SD rats in each group of Example 3; among them, Figure 1 The normal group Figure 2 Model group, Figure 3 All-round group, Figure 4 Moxibustion acupuncture group.

[0026] Figure 5-8 Masson staining images (10.0x) of liver tissue sections from SD rats in each group of Example 3; among them, Figure 5 The normal group Figure 6 Model group, Figure 7 All-round group, Figure 8 Moxibustion acupuncture group.

[0027] Figure 9 Example 5 shows the detection results of serum hyaluronic acid, type IV collagen, and type III procollagen in each group. In the figure, HA: serum hyaluronic acid; IV: type IV collagen; III: type III procollagen.

[0028] Figure 10 Example 5: Detection results of AST, ALT, and ALP in each group.

[0029] Figure 11 Example 5: Detection results of TBIL, DBILI, and IBIL in each group.

[0030] Figure 12 Example 5: TBA detection results for each group. Detailed Implementation

[0031] To better understand the technical content of this invention, specific embodiments are provided below to further illustrate the invention.

[0032] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0033] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0034] Example 1

[0035] Drug composition: 60g of dried aerial parts of Ginger sibthorpioides, 20g of dried aerial parts of Artemisia argyi, and 30g of whole Bidens pilosa.

[0036] Example 2

[0037] Drug composition: 50g of dried aerial parts of Ginger sibthorpioides, 22g of dried aerial parts of Artemisia argyi, and 35g of whole Bidens pilosa.

[0038] Example 3

[0039] Drug composition: 70g of dried aerial parts of Ginger sibthorpioides, 18g of dried aerial parts of Artemisia argyi, and 25g of whole Bidens pilosa.

[0040] Example 4

[0041] Drug composition: 60g of dried aerial parts of Ginger sibthorpioides, 20g of dried aerial parts of Artemisia argyi, and 30g of whole Bidens pilosa.

[0042] Preparation method: Ginger sibthorpioides (dried aerial parts), Artemisia argyi (dried aerial parts), and Bidens pilosa (whole herb) were pulverized separately and weighed according to their respective weights. They were then soaked in water for 30 minutes, with the water covering the herbs by about 2 cm. The mixture was decocted until boiling and kept boiling for 1 hour. After filtration, the residue was boiled again for 30 minutes. The filtrates were collected twice, concentrated, and freeze-dried to obtain dry powder.

[0043] Comparative Example 1

[0044] Drug composition: 20g of dried aerial parts of Artemisia argyi and 30g of whole Bidens pilosa.

[0045] Preparation method: Gynostemma pentaphyllum (dried aerial parts) and Bidens pilosa (whole herb) were pulverized separately and weighed according to the above-mentioned weights. They were soaked in water for 30 minutes, with the water covering the herbs by about 2 cm. The mixture was boiled and kept boiling for 1 hour. After filtration, the residue was boiled again for 30 minutes. The filtrates were collected twice, concentrated, and freeze-dried to obtain dry powder.

[0046] Example 5

[0047] 1.1 Experimental Methods

[0048] Forty SD rats were randomly divided into four groups: a model group, a complete formula group, an acupuncture group, and a normal control group, with 10 rats in each group. After one week of acclimatization and observation, a rat HF ​​model was established by intraperitoneal injection of carbon tetrachloride (CCl4). CCl4 was dissolved and diluted with olive oil, and a 40% CCl4 solution was prepared by mixing olive oil and CCl4 at a volume ratio of 3:2. This solution was prepared fresh before each intraperitoneal injection. Rats in each group were weighed and their weight recorded before each intraperitoneal injection. The first intraperitoneal injection was 0.4 mL / 100g body weight of the 40% CCl4 solution, followed by 0.2 mL / 100g, twice a week (Monday and Thursday) for 8 weeks. The normal control group received the same volume of olive oil.

[0049] Administer the medication starting in week 5 of modeling and continue for 4 weeks. Dosage: 1 ml / 100g body weight.

[0050] The complete formula group: Take the drug (dry powder) prepared in Example 4, dilute it with physiological saline and administer it to the drug, with a dilution concentration of 0.5g / ml.

[0051] Moxibustion group: Take the drug (dry powder) prepared in Comparative Example 1, dilute it with physiological saline and administer it to the patient. The dilution concentration is 0.5g / ml.

[0052] The day after the last administration, blood was drawn from the abdominal aorta under chloral hydrate anesthesia. The blood was centrifuged, and serum was collected for various liver function tests. Serum hyaluronic acid, type III procollagen, and type IV collagen were measured using an ELISA kit. Liver tissue was then taken for pathological examination after the blood draw.

[0053] 1.2 Experimental Results

[0054] Depend on Figure 1-4 It can be seen that, compared with the normal group, the model group rats exhibited disrupted liver lobule structure, with extensive proliferation of fibrous tissue in the portal area forming fibrous cords that segmented and surrounded the liver lobules to form pseudolobules. Hepatocyte cords were disordered, with widespread fatty degeneration, partial hepatocyte necrosis, and inflammatory cell infiltration within the fibrous septa. Compared with the model group, the disordered liver lobule structure in the whole-treatment group was significantly improved, with orderly arrangement of hepatic cords, significantly reduced fibrous tissue proliferation, and significantly reduced hepatocyte degeneration and inflammatory cell infiltration. The liver damage in the moxibustion group was improved compared with the model group, but the damage was still significant compared with the whole-treatment group.

[0055] Depend on Figure 5-8 It can be seen that, compared with the normal control group, the portal area of ​​the liver tissue of rats in the model group was enlarged, with a large amount of collagen fiber deposition and obvious fibrous septa; compared with the model group, the collagen fiber deposition and fibrous septa of the liver tissue of rats in the whole formula group were significantly reduced; compared with the whole formula group, the collagen fiber hyperplasia of the liver tissue of rats in the moxibustion group was still very obvious.

[0056] Table 1. Detection results of HA, IV, and III in serum of each group (ng / ml)

[0057] normal group 10 86.1 28.1 14.7 Model group 10 245.7*** 69.1** 35.3** All-round group 10 107.5 ### ]]> 38.5 ## ]]> 20.1 # ]] Moxibustion acupuncture group 10 173.3 ## ]]> 47.6 # ]]> 26.9

[0058] Compared with the normal group, *P<0.05, **P<0.01, ***P<0.001; compared with the model group, # P<0.05, ## P<0.01, ## P < 0.001.

[0059] Colloidal type IV (CtIV) is distributed beneath the sinusoidal endothelial cells of the liver and is a major component of the liver basement membrane. After being synthesized intracellularly, it directly participates in the formation of the extracellular matrix using procollagen. Type III collagen (PCIII) reflects the synthesis of type III collagen in the liver. Serum PCIII levels are closely correlated with the degree of liver fibrosis and are of significant value in the early diagnosis of liver fibrosis. Hepatocellular carcinoma (HA), a matrix component synthesized by interstitial cells, can accurately and sensitively reflect the amount of fibrosis already formed in the liver and the state of hepatocyte damage, making it a sensitive indicator of liver fibrosis and cirrhosis. Therefore, CtIV, CtIII, and HA can all be used as indicators to measure the degree of liver fibrosis. Increased or decreased levels indicate either a worsening or slowing of the progression of liver fibrosis.

[0060] Depend on Figure 9 As shown in Table 1, compared with the normal group, the model group showed significantly elevated levels of HA, IV, and III. Compared with the model group, the whole formula group and the moxibustion group showed significantly decreased levels of all three. Compared with the moxibustion group, the whole formula group showed a more significant reduction in HA, IV, and III levels. This indicates that the whole formula group has a more significant therapeutic effect on improving liver fibrosis.

[0061] Table 2 shows the detection results (U / L) of AST, ALT, and ALP in each group.

[0062] normal group 10 168.2 55.0 303.2 Model group 10 495.1*** 193.6*** 358.3* All-round group 10 305.0 ## ]] 112.7 ## ]]> 281.4 # ]]> Moxibustion acupuncture group 10 385.9 # ]]> 158.7 # ]]> 316.7

[0063] Compared with the normal group, *P<0.05, **P<0.01, ***P<0.001; compared with the model group, # P<0.05, ## P<0.01, ## P < 0.001.

[0064] ALT and AST are important indicators of liver parenchymal damage; elevated levels indicate hepatocyte damage, and the greater the elevation, the more severe the damage. ALP is an important enzyme for the excretion of bile acids in the liver; when the liver is damaged or the bile ducts are blocked, alkaline phosphatase levels will increase.

[0065] Depend on Figure 10 As shown in Table 2, compared with the normal group, the levels of AST, ALT, and ALP in the model group were significantly increased, indicating significant hepatocellular damage. After intervention with the whole-formula group and the moxibustion group, the levels of AST, ALT, and ALP all decreased significantly, indicating that both could alleviate hepatocellular damage and reduce liver enzymes. Compared with the moxibustion group, the whole-formula group showed a more significant decrease in AST, ALT, and ALP levels, demonstrating better liver protection and enzyme-lowering effects.

[0066] Table 3 shows the detection results (umol / L) of TBIL, DBILI, and IBIL for each group.

[0067] normal group 10 0.6 1.3 0.5 Model group 10 4.4*** 3.3** 1.2** All-round group 10 <![CDATA[2.5 ## ]]> <![CDATA[1.8 # ]]> <![CDATA[0.7 # ]]> Moxibustion acupuncture group 10 <![CDATA[3.3 # ]]> 2.9 1.0

[0068] Compared with the normal group, *P<0.05, **P<0.01, ***P<0.001; compared with the model group, # P<0.05, ## P<0.01, ## P < 0.001.

[0069] Bilirubin is classified into indirect bilirubin (IBIL), direct bilirubin (DBILI), and total bilirubin (TBIL). Indirect bilirubin originates from hemoglobin released by red blood cells and is taken up by liver cells and converted into direct bilirubin. When liver function is impaired and the liver is unable to effectively take up and convert bilirubin, or when liver fibrosis causes compression of intrahepatic bile ducts, obstructing bile excretion, the levels of TBIL, DBILI, and IBIL increase.

[0070] Depend on Figure 11 As shown in Table 3, compared with the normal group, the levels of TBIL, DBILI, and IBIL in the model group were significantly increased. Compared with the model group, the levels of TBIL, DBILI, and IBIL in the whole formula group and the moxibustion group were significantly decreased. Compared with the moxibustion group, the whole formula group showed a more significant reduction in the levels of TBIL, DBILI, and IBIL. This indicates that the whole formula group can better protect hepatocytes, reduce liver damage, and decrease liver fibrosis.

[0071] Table 4. Detection results of TBA in each group

[0072] normal group 10 17.2 Model group 10 81.5*** All-round group 10 <![CDATA[68.2 # ]]> Moxibustion acupuncture group 10 73.6

[0073] Compared with the normal group, *P<0.05, **P<0.01, ***P<0.001; compared with the model group, # P<0.05, ## P<0.01, ## P < 0.001.

[0074] Depend on Figure 12 As shown in Table 4, TBA levels in the model group were significantly higher than those in the normal group. After comprehensive intervention, TBA levels decreased significantly, while there was no significant difference in the moxibustion group. This indicates that the comprehensive intervention group can better reduce liver fibrosis, alleviate intrahepatic bile duct obstruction, and facilitate bile excretion.

[0075] In summary, compared with the moxibustion group, the whole formula group can better reduce the levels of HA, IV, III, AST, ALT, ALP, TBIL, DBILI, IBIL, and TBA. The whole formula group is more effective in reducing liver damage and improving liver fibrosis.

[0076] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A traditional Chinese medicine composition for improving liver fibrosis, characterized in that, It is made from the following raw materials in the following weight ratios: 50-70g of *Gnaphalium affine*, 18-22g of *Artemisia argyi*, and 25-35g of *Bidens pilosa*; the *Gnaphalium affine* being the dried aerial part of *Gnaphalium affine*.

2. The traditional Chinese medicine composition for improving liver fibrosis according to claim 1, characterized in that, It is made from the following raw materials in the following weight ratio: 55-65g of ginger, 18-22g of Artemisia argyi, and 25-35g of Bidens pilosa.

3. The traditional Chinese medicine composition for improving liver fibrosis according to claim 1, characterized in that, It is made from the following ingredients in the following weight ratio: 60g of ginger, 20g of artemisia, and 30g of beggar-ticks.

4. The traditional Chinese medicine composition for improving liver fibrosis according to claim 1, characterized in that, The *Artemisia argyi* mentioned refers to the dried aerial parts of *Artemisia argyi*.

5. The traditional Chinese medicine composition for improving liver fibrosis according to claim 1, characterized in that, The Bidens pilosa mentioned refers to the whole plant of Bidens pilosa.

6. A traditional Chinese medicine composition for improving liver fibrosis, characterized in that, It is prepared from the traditional Chinese medicine composition according to any one of claims 1 to 5.

7. A method for preparing the traditional Chinese medicine composition according to any one of claims 1 to 5, characterized in that, Includes the following steps: Take the herbs of *Gnaphalium affine*, *Artemisia argyi*, and *Bidens pilosa*, crush them, add water and soak for 20-40 minutes, then boil and extract for 0.8-1.2 hours. Filter, add water to the residue and continue to boil and extract for 20-40 minutes. Collect the two filtrates, concentrate and freeze dry to obtain dry powder.

8. The use of the traditional Chinese medicine composition according to any one of claims 1 to 5 or the traditional Chinese medicine composition prepared by the preparation method according to claim 7 in the preparation of a drug for treating liver fibrosis.

9. The application according to claim 8, characterized in that, The application of the traditional Chinese medicine composition in the preparation of a drug for treating liver fibrosis of the dampness-blood stasis type.

Images