WO2006026926A1

WO2006026926A1 - A medicinal composition for treating hepatic fibrosis and its preparation

Info

Publication number: WO2006026926A1
Application number: PCT/CN2005/001439
Authority: WO
Inventors: Hongwu Zhang; Qingjuan Li; Cuiyan Liu; Min Liang; Rongduan Wang; Lifang Wang; Qian Zhao
Original assignee: Shijiazhuang Pharma. Group Zhongqi Pharmaceutical Technology (Shijiazhuang) Co., Ltd.
Priority date: 2004-09-10
Filing date: 2005-09-09
Publication date: 2006-03-16
Also published as: CN1308028C; CN1726999A

Abstract

The present invention relates to a medicinal composition for treating hepatic fibrosis and its preparation The composition is maily prepared by using the Chinese medicinal materials of 10-30 weight portions of Radix Gentianae, 20-40 weight portions of Rhizorna Curcumae, 30-50 weight portions of Radix Scutellariae, 15-35 weight portions of Rhizoma Sparganii, 25-45 weight portions of Radix Saluiae Miltiorrhizae,15-35 weight portions of Rhizoma Atractylodis Macrocephalae, 10-30 weight portions of Radix Astragali. It has effects for treating hepatic fibrosis and early cirrhosis.

Description

Medicine composition for treating liver fibrosis and preparation method thereof

The present invention relates to a pharmaceutical composition for treating liver fibrosis and a preparation method thereof.

Background technique

Liver fibrosis belongs to the name of Western medicine. As far as its clinical manifestations are concerned, it is mainly attributed to the category of "hypochonism", "symptoms", "products," and so on.

In terms of "hypochonism": "Nei Jing" has "the evil is in the liver, then the two threats are painful... the evil blood", "the evil lover is in the foot of the Shaoyang, it is not a threat." The theory of interest. Zhang Jingyue, a doctor of the Ming Dynasty, divided the "hypochondriac pain" into two categories: exogenous and internal injuries, and clearly pointed out that "...but the internal pain and the painful person are ten or eight, and there is an ear between the exogenous hypochondriac." "Su Wen. The theory of dirty air": "The liver disease, the pain under the two threats ..."; "Certificate of the syndrome" pointed out the cause of the disease ... ... or the death of blood stasis , or angry stagnation of liver and fire attack ... are all painful flank pain."

In terms of "symptoms" and "products": "Difficult Sutra" said: "There are accumulated people, the five internal organs are born...", "Golden 匮 · · 五五五风风风风风五五五五五五五五五五五五: "The accumulators, the ills are also, the stagnation is not moving; the gatherers, the rickets are also; the seizures sometimes, the pain of the exhibition, can be treated, ...". "All kinds of diseases and syndromes": "The symptoms are all caused by cold and cold, and the diet is not changed. It is also born with the organs. The disease is directly called the disease.瘕瘕 , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , Most of the doctors of the past dynasties argued from emotional disorders, eating injuries, and feeling the evils. For example, "The Golden Chamber, Accumulation of the General Theory" pointed out that "whoever is worried and angered, long-term unsolvable, and more into this disease.""Jing Yue Quan Shu" pointed out that "the stagnation of diet, staying in the middle ... not to be able to do, to block, is the accumulation.""Lingshu" pointed out that "the beginning of the accumulation, the birth of the child" The above various explanations indicate that there are many causes, but the etiology is multi-faceted. The main cause is qi stagnation and blood stasis, and the heat and turbidity are all contributing to qi stagnation. Bloody factors.

It should be noted that the formation of accumulation is closely related to the strength of the righteousness. As early as in the "Nei Jing", there is "the brave man is arrogant, and the deaf is the disease."

The motherland medicine also has a lot of arguments about its etiology. It is believed that due to emotional disorders, unhealthy food and drink, the feeling of wetness and phlegm, and chronic illness, or jaundice, the viscera and qi and blood function are impaired, resulting in air-conditioning. Block, stop bleeding. "Zhengzhi Huibu" puts forward the cause of "causal anger, sorrow, sorrow, overeating, cold, sorrow, sorrow, stagnation, stagnation, or blood stasis" Pain, as for the hot and humid smoldering fire, the servant room color and the sick, there are also.""Golden 匮 · · 积积积积》》 : : : : : : : 凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡凡郁凡》: "Damp and heat intersect, the people are sick and sick";"Ge to Yu theory · bulging theory": "This is also the seven emotions of internal injuries, the six genocide, the diet is not good, ... 遂成满满, In the stage of chronic hepatitis, it is often due to unclean diet, wet and hot sinister poisoning; Injury of the spleen and stomach, loss of spleen, loss of power, heat and phlegm; or emotional depression, so that the liver is not comfortable, spleen qi stagnation, leading to liver and spleen qi stagnation, followed by gas and blood, making blood flow poor , 脉瘀。。. If the damp heat stays for a long time, then the righteous gas is depleted, the temper is declining, the righteousness is not good, the turbidity is not, and the wet turbidity is condensed and cemented. "Danxi Heart Law - Bulging" pointed out: "Seven love internal injuries, six genocide, eating yin and yin injury, the transfer of the official dereliction of duty, although the stomach is affected by the valley, can not be transported, the self-rising of the sun, Yin Drop, the world is not handed over, the turbidity is mixed, the tunnel is blocked, the stagnation is hot, the heat stays wet, the damp heat is born, the ^成^ I is full, the bulging is also "the medical entry" bulging 》: "Inflated yin and cold is evil, ... the real yang heat is evil, ... so the turbidity is in the lower, turned into bloody, the stagnation is hot, the heat becomes wet, the damp heat fights, the swells bulge" . Damp-heat internal resistance, prolonged course of disease, will further affect the operation of blood, resulting in stagnation of blood, blood stasis, blood stasis and phlegm dampness, and for a long time, it will become a tangible product, and under the ribs, it will become a sputum. Block (hepatosplenomegaly), condensed hard, pushes it, Yu Chang: "Yin Qi does not disperse", "Water wraps and blood stasis", if the pulse is blocked, Then the collaterals are angered, and the blue veins are exposed (venous anger).

Traditional Chinese medicine has accumulated a lot of experience in the treatment of this disease in the past. For example, "Ancient and Modern Medical Journal" puts forward "... When the rule is to disperse qi and phlegm and blood, and flatten its liver and lead its qi, Then there is no cure." In the "Zhangshi Medical Communication", the discussion is more detailed, and the following: "...Li Shicai's succession is also based on the accumulation of the product, and the lack of righteousness, and then the evil spirits, but the attack is too urgent and righteous, junior high school The last three laws must not be said. The first person is sick and the beginning of the disease... then he is attacked, the middle person... is subject to attack and compensation, and the last is... To be compensated. ...... "; Gu Yun 'strong people have no product, but the virtual person has it. ... good living. When you first make up the imaginary, make the blood strong, and the accumulation is also eliminated. Do not ask what is dirty, first adjust it, enable the diet, it is also the original. ...... If the husband is a big gathering, don’t search for it, and the Japanese will make up for it, and it will be no good, ...... send the soldiers into the army to discuss it, why not suffer from it." Reading also said: "Being sick for a long time, the evil spirits are deeper, the righteousness is weakened, and it is subject to attack and compensation."

Therefore, according to the etiology and pathogenesis of liver fibrosis and the treatment of ancient literature, it is believed that the pathogenesis of liver fibrosis and early liver cirrhosis is dampness and heat intrinsic, blood stasis, qi deficiency and spleen weakness. The disease is located in the liver and spleen, and the disease is virtual, dampness and blood stasis is the key to the disease. According to the above analysis, the rule of removing the damp heat and removing the stagnation of the tangible congestion is also established, and the spleen and qi are strengthened. Under the guidance of this theory, people are unanimously looking for drugs with better curative effect on liver fibrosis.

Summary of the invention

The invention is mainly for clearing heat, dissipating phlegm and eliminating phlegm, and taking the spleen and qi as the treatment rule, so that the evil of the liver and gallbladder is clear, the blood stasis is eliminated, the spleen and stomach function is restored, the spleen is spleen, and the protection is achieved. The purpose of the liver. The present inventors have obtained a large number of animal and clinical trials, and have screened a Chinese medicine composition with a positive effect, that is, a clear liver and phlegm medicine composition, thereby completing the present invention.

SUMMARY OF THE INVENTION A primary object of the present invention is to provide a pharmaceutical composition for treating liver fibrosis, followed by a method of preparing the pharmaceutical composition.

- The present invention provides a pharmaceutical composition for treating liver fibrosis, also known as liver and phlegm medicine, It should be made from the following APIs: gentian, zedoary, radix scutellaria, striatum (vinegar), salvia miltiorrhiza, atractylodes, scutellaria, and their parts by weight: 10 to 30 parts of gentian, 20 parts of medlar (vinegar) 30 50 parts of Astragalus, 15 ~ 35 parts of Sanling (vinegar), 25 ~ 45 parts of Salvia miltiorrhiza, 15 ~ 35 parts of Atractylodes, 10 ~ 30 parts of Astragalus.

The pharmaceutical composition of the present invention may further comprise tangerine peel and Polygonum cuspidatum, and their parts by weight are:

10 ~ 30 servings, and 10 to 30 copies of Polygonum cuspidatum.

The pharmaceutical composition of the present invention may further contain licorice in a weight fraction of 5 to 15 parts.

The pharmaceutical composition of the present invention can be prepared into any conventional oral dosage form by a conventional method of a traditional Chinese medicine preparation, but in order to make the raw material Chinese herbal medicine in the medicine exert a better pharmacological effect, it is necessary to extract the active ingredient from the medicine.

When the pharmaceutical composition of the present invention contains at least gentian, medlar, radix, scorpion (vinegar), salvia, atractylodes, and astragalus, the preparation method is as follows:

a, 莪 ( (vinegar system), atractylodes water vapor steam extraction method, collect volatile oil, spare, volatile oil can also be used after inclusion of 3-cyclodextrin, the distilled aqueous solution is collected;

b, Astragalus, Triangular (vinegar), Astragalus, Dan participate in water reflux extraction, combined extract, filtered, the filtrate is combined with the aqueous solution collected in step a, concentrated, ethanol, refrigerated overnight, filtered, supernatant used ;

• c, gentian and ethanol are heated to reflux, the extract is filtered, the filtrate is combined with the supernatant prepared in step b, and the ethanol is recovered to an alcohol-free taste and concentrated;

d. The pharmaceutical composition of the present invention is obtained by combining the j8-cyclodextrin inclusion compound of the volatile oil or the volatile oil prepared in the step a and the concentrate prepared in the step c.

The concentrate in step c is preferably in the form of a clear paste and is preferably spray dried into a dry extract powder.

When the pharmaceutical composition of the present invention further contains tangerine peel and Polygonum cuspidatum, the preparation method is as follows: a, zedoary (vinegar), atractylodes and tangerine peel extracted by steam distillation, collecting volatile oil, spare, volatile oil can also be used after /3-cyclodextrin inclusion, the distilled aqueous solution is collected; b, jaundice, three Ribs (vinegar), Polygonum cuspidatum, Astragalus membranaceus, Dan, participate in water reflux extraction, combine the extracts, filter, and combine the filtrate with the aqueous solution collected in step a, concentrate, add ethanol, 'refriger overnight, filter, and use the supernatant;

c, gentian and ethanol are heated to reflux, the extract is filtered, the filtrate is combined with the supernatant prepared in step b, and the ethanol is recovered to an alcohol-free taste and concentrated;

d. The 3-cyclodextrin inclusion complex of the volatile oil or volatile oil prepared in the step a and the concentrate prepared in the step c are combined to obtain the pharmaceutical composition of the present invention.

When the pharmaceutical composition of the present invention further contains licorice, the preparation method is as follows: a, sputum (vinegar), atractylodes and tangerine peel are extracted by steaming, collecting volatile oil, and the volatile oil can also be wrapped with cyclodextrin. After the standby, the distilled aqueous solution is collected separately; b, Astragalus, Sanling (vinegar), Polygonum cuspidatum, Astragalus, Salvia miltiorrhiza, licorice and water reflux extraction, combined extract, filtered, and the filtrate is combined with the aqueous solution collected in step a, concentrated , add ethanol, refrigerate overnight, filter, and use the supernatant for use;

In the treatment of liver fibrosis medicine of the present invention, each of the preparations required for preparing different dosage forms can be added. Conventional excipients, such as disintegrants, lubricants, binders, etc., are prepared into any of the commonly used oral dosage forms, such as pills, powders, tablets, capsules, oral solutions, and the like, by conventional Chinese medicine preparation methods.

Analysis:

The party uses gentian and phlegm as the medicinal herbs. The former clears the liver and gallbladder dampness heat, and the latter ruptures the blood and eliminates the qi. "For the liver and blood, the medicine can break the blood in the gas, and the two drugs can be used to clear the liver." Effect: Huangqi Fulong gentian clear liver and dampness, Sanling assisted phlegm surgery to eliminate phlegm and phlegm; with Danshen activating blood circulation and phlegm and collaterals, there is a "breaking blood, new blood" wonderful; with Huang Qi, Bai Shu Jian The spleen and qi, Fuzheng Pei Ben, the elixir drug to the evil spirits and not to hurt the positive, the above five flavors are a common medicine, and the medicine is matched with the liver and phlegm, the evil spirits help the right. The tiger stick can be the medicine of the monarch, one can heat Lishui, two can activating blood circulation to remove phlegm; Chenpi with its spleen and stomach power can make the medicinal herbs "smell evil without hurting the positive,, with its effect of qi and dampness, so that the medicinal herbs Astragalus, Atractylodes" Convergence, and the two are adjuvants. The licorice is slow, and the sacred medicine is the sin of the sin, and the stipulations of the stipulations of the medicinal herbs, and the various medicines can be adjusted. The versatile compatibility, the loyalty of the princes, the order Clearing the liver and dehumidifying, activating blood circulation and removing blood stasis, strengthening the spleen and replenishing qi. When the damp heat goes, the sputum is smooth; Jian, it is strong. For the cirrhosis, the tiredness of the body, the complexion is dull, the yellow body is yellow, the liver and spleen is swollen, the abdomen is tired or the two ribs are painful, and the mouth is bitter and stagnation. Therapeutic effect.

The function of the clear liver and phlegm medicine composition of the invention is Qinggan Huatan, Jianpiyiqi.

Adaptation range: For the treatment of liver fibrosis and early cirrhosis, Zheng Jian liver and gallbladder damp heat, liver blood stasis, spleen weak qi deficiency caused by dull complexion, yellow body yellow, abdominal distension or two flaccid pain, postprandial aggravation The mouth is bitter, the body is tired, the urine is red, the tongue is dark purple, there is electricity or freckle, the tongue is yellow, the pulse string is thin or the string is slippery.

detailed description

The preparation method of the pharmaceutical composition of the present invention is further illustrated by the following examples.

Example 1: Preparation of a pharmaceutical composition for treating liver fibrosis (Qinggan Huayu Drug)

1. Raw material formula: 20 parts of gentian, 30 parts of medlar (vinegar), 40 parts of astragalus, ternary (vinegar) 25), 35 parts of Salvia, 25 parts of Atractylodes, 20 parts of Astragalus, 20 parts of dried tangerine peel, 20 parts of Polygonum cuspidatum, 10 parts of licorice.

2, preparation method:

a, sputum (vinegar), atractylodes and tangerine peel plus 8 times the amount of water, extracted by steam distillation for 6 hours, collect the volatile oil, after inclusion with /3-cyclodextrin for use, the distilled aqueous solution is collected; b, Astragalus, Sanling (vinegar), Polygonum cuspidatum, Astragalus, Salvia miltiorrhiza, licorice and 10 times water were extracted three times (2 hours each time), the extracts were combined, filtered, and the filtrate was combined with the aqueous solution collected in step a, concentrated under reduced pressure. To 1: 1, add ethanol to make the alcohol content up to 60%, refrigerate overnight, filter, and use the supernatant for use;

c. The gentian is heated and refluxed with 70% ethanol three times (one hour each time), the extracts are combined, filtered, and the filtrate is combined with the supernatant prepared in step b, the ethanol is recovered to an alcohol-free taste, and concentrated into a clear paste, sprayed Dry to dry extract powder;

d. Combining the 3-cyclodextrin inclusion compound of the volatile oil prepared in the step a with the dry extract powder prepared in the step c, the pharmaceutical composition for treating liver fibrosis of the present invention is obtained.

Example 2: Preparation of hard capsules for treating liver fibrosis drugs

Preparation of a capsule preparation on the basis of Example 1

Example 1 Pharmaceutical Composition 387g

Micro-silica gel 10g

Stearic acid touch 2g

Mix the parts of the above formula and fill them to make 1000 capsules. 0.4g per capsule, 15 capsules a day.

Example 3: Preparation of soft capsules for treating liver fibrosis drugs

Preparation of a capsule preparation on the basis of Example 1

The pharmaceutical composition obtained in Example 1 was 383 g Medicinal soybean oil 258g

Glyceryl monostearate 0.4g

Take 1 part of gelatin, add 1 part of water, 0.6 parts of glycerin, melt at 70-80 °C, transfer to 55-65 °C in a heat preservation tank for use, take medicinal soybean oil, glyceryl monostearate, heat, The solution was mixed at 70 ° C, allowed to cool, and the active component of the above-mentioned liver-clearing and phlegm-removing drug was added, stirred and mixed, and ground with a colloid mill to form a uniform suspension. The prepared chemical solution was added to a supply tank for pelletization to prepare 1000 tablets. Soft gelatin is set at 25 °C and 30% relative humidity for 20 hours, then transferred to a dryer for drying. The drying conditions are 30 ° C, the relative humidity is 20%, the wind speed is 0.6 m / s, and the time is 8 h. . After drying, the soft plastic bottle is washed, washed, and washed. 0.65g per capsule, 12 capsules per day.

Example 4: Preparation of a pharmaceutical composition for treating liver fibrosis (Huangan Huayu Drug)

1. Raw material formula: 26 parts of gentian, 32 parts of medlar (vinegar), 32 parts of astragalus, 25 parts of trigonal (vinegar), 35 parts of salvia, 25 parts of atractylodes, 20 parts of astragalus.

2, preparation method

a, 莪 ( (vinegar system;), Atractylodes plus 8 times the amount of water, extracted by steam distillation for 6 hours, collect the volatile oil, after inclusion with 5-cyclodextrin for use, the distilled aqueous solution is collected; b, jaundice, Triangular (vinegar), Astragalus, and Dan were added to 10 times the amount of water and refluxed three times (2 hours each time). The extracts were combined, filtered, and the filtrate was combined with the aqueous solution collected in step a, and concentrated under reduced pressure to 1:1. Ethanol is made up to 60% alcohol, refrigerated overnight, filtered, and the supernatant is ready for use;

c. The gentian is heated and refluxed three times with 70% ethanol (1 hour each time), and the extracts are combined, filtered, and the filtrate is combined with the supernatant prepared in step 3, and the ethanol is recovered to an alcohol-free taste, and concentrated into a clear paste. Spray drying into a dry extract;

d. The cyclodextrin inclusion compound of the volatile oil prepared in the step a and the dry extract powder prepared in the step c are combined to obtain the therapeutic composition for treating liver fibrosis of the present invention. Example 5: Preparation of a hard capsule for treating liver fibrosis

Preparation of a capsule preparation on the basis of Example 4

Example 4 Pharmaceutical Composition 356g

Lactose 30g

Micro-silica gel 12g

Magnesium stearate 2g

The parts of the above prescription are mixed and filled, and 1000 capsules of glue are obtained. 0.4g per capsule, 15 capsules per serving.

Example 6: Preparation of a pharmaceutical composition for treating liver fibrosis (Huangan Huayu Drug)

1. Raw material formula: 30 parts of gentian, 40 parts of medlar (vinegar vinegar), 40 parts of astragalus, 30 parts of ternary (vinegar vinegar), 40 parts of salvia miltiorrhiza, 25 parts of atractylodes, 25 parts of astragalus, 25 parts of dried tangerine peel, 25 parts of Polygonum cuspidatum .

2, preparation method:

a, sputum (vinegar), atractylodes and tangerine peel plus S times water, steam extraction method for 6 hours, collect the volatile oil, after inclusion with /3-cyclodextrin, the distilled aqueous solution is collected; b , Astragalus, Triangular (vinegar), Polygonum cuspidatum, Astragalus, and Dan participated in 10 times water reflux extraction three times

(2 hours each time), the extracts were combined, filtered, and the filtrate was combined with the aqueous solution collected in step a, concentrated under reduced pressure to 1 1 , ethanol was added to make the alcohol content 60%, refrigerated overnight, filtered, and the supernatant was used. c. The gentian is heated and refluxed with 70% ethanol twice (1 hour each time), the extracts are combined, filtered, and the filtrate is combined with the supernatant prepared in step b, and the ethanol is recovered to an alcohol-free taste and concentrated into a clear paste. Spray drying into a dry extract powder;

d. Combining the 3-cyclodextrin inclusion compound of the volatile oil prepared in the step a with the dry extract powder prepared in the step c, the pharmaceutical composition for clearing liver and phlegm of the present invention is obtained.

To illustrate the effect of the pharmaceutical composition of the present invention on liver damage, the therapeutic effect on liver fibrosis was examined by establishing an animal model. Experiment 1. Effect of capsules (pharmaceutical composition for treating liver fibrosis) on immunosuppressive liver fibrosis in rats

Test purpose

Intravenous bovine serum albumin was used to replicate the rat model of immune-induced liver fibrosis, and the therapeutic effect of Qinggan Huayu Capsule was observed.

2 test materials

2.1 drugs

The liver phlegm and hard sputum prepared according to Example 2, the powder was reddish brown, and was mixed with a 0.5% aqueous solution of carboxyhydrazine-based cellulose to prepare a suspension of a suitable concentration.

Compound 鳖曱soft liver tablets: Commercially available, batch number 20021010, used with a 0.5% carboxymethyl cellulose aqueous solution to prepare a suspension of appropriate concentration.

Colchicine: Serva's product, batch number C9754, is used in a solution of the appropriate concentration in heavy distilled water.

2. 2 reagents

Bovine serum albumin (BSA): Tianjin Haoyang Biological Products Co., Ltd., batch No. A-7030 Lanolin: Shanghai Jiading Huating Lanolin Factory, batch number 000917

Liquid Paraffin: Tianjin Chemical Reagent Second Factory, Batch No. 980425

Freund's incomplete adjuvant: 120g of lanolin added to liquid paraffin 240ml, stirred and mixed in a 65 °C water bath, autoclaved and stored in a refrigerator at 4 °C.

Coomassie brilliant blue protein assay kit: Nanjing Institute of Bioengineering, batch number

20030715

Hydroxyproline Assay Kit: Nanjing Jianshe Bioengineering Institute, Lot No. 20030715 Sialic Acid Assay Kit: Nanjing Jiansheng Bioengineering Institute, Lot No. 20030707 2.3 Animals Rat: Wistar species, secondary animal, provided by the Fourth Academy of Military Sciences, Certificate No.: Military Medical B98014

Feeding conditions: caged, 5-6 per cage, free drinking water, feeding, room temperature controlled by central air conditioning at 24 ± 2 ° C, humidity 50 ± 15%, light 12 hours bright, 12 hours dark (6 am Bright, '6 pm dark.

3 test method

3.1 Model replication

100 healthy Wistar rats were used, weighing 130-150g, male and female, except for the normal control group (5 males and 5 females), the other 90 rats were treated with 9mg/ml BSA Freund's incomplete adjuvant suspension. Each time 0.5 ml/head, multiple subcutaneous injections were performed to sensitize the animals for 5 consecutive times. The first and second intervals are 2 weeks, and the rest are separated by 1 week. One week after the last sensitization, all animals were bled in the orbital vein and serum BSA antibodies were determined by capillary method. Take the antibody-positive animal tail vein and inject different concentrations of BSA physiological saline solution 0.4ml / 2 times a week, the dose from 2.0mg / 0.4ml / only, sequentially increased to 3.0mg / 0.4ml / only (increment O.lmg), the 12th time was 3.4mg/0.4ml/only, and the subsequent increments were 0.2mg to 4.0mg/0.4ml/only for 15 times. The normal group rats were treated with normal saline instead of BSA for tail vein injection. Three days after the last attack, blood was taken to determine the ratio of ALT, AST, TP, ALB, A/G (albumin/globulin), sialic acid; part of the liver was taken, and the content of hydroxyproline was determined after homogenization; The liver was fixed with 10% formalin, HE staining and Masson staining, and the degree of liver fibrosis was observed under light microscope.

3.1.2 grouping method

Except the normal control group, the other animals were randomly divided into 6 groups according to body weight after 4 times of challenge injection. Model control group, Qinggan Huayu capsule 嚢 0.9, 1.8, 3.6g powder/kg dose group, positive drug compound 鳖曱Soft liver tablets 2g powder / kg dose group, positive drug colchicine 0.25mg / kg dose group, plus the normal control group, a total of 7 groups. 3.1.3 administration time

All the animals in each group were intragastrically administered with a perfusion volume of 1 ml/100 g body weight after 4 times of challenge injection. The normal control group and the model control group were intragastrically administered with an equal volume of 0.5% carboxymethyl cellulose suspension. 1 time, 7 times a week for 6 consecutive weeks.

3.1.4 Data Statistics

The test data is expressed as i±SD, and the two groups are statistically analyzed by the 4-value method, and the rank-sum test is used for statistical analysis.

4 test face results

4.1 General situation

After 5 times of sensitization, a total of 87 rats with positive serum BSA antibody (3 male rats did not meet the test requirements), when the attack was injected, the rats developed different degrees of allergic shock-like reactions, shortness of breath, and no walking. Stable, prone, and even death, the dead animals were immediately dissected, and no obvious abnormalities were observed by the naked eyes. The surviving animals returned to normal in about 30 minutes. Most of the dead animals appeared in the first 1-3 attacks, and from the fourth attack, occasionally, the animals developed an allergic shock-like reaction. After 4 injections, 16 deaths (10 males and 6 males) and 71 surviving animals were randomly divided into 6 groups according to body weight. Model control group, Qinggan Huayu 0.9g powder/kg dose group Qinggan Huayu 1.8g powder / kg dose group, Qinggan Huayu powder 3.6g powder / kg dose group, compound Biejia soft liver tablets 2g powder / kg dose group each group of 12 (female, male each 6 ), 11 colchicine 0.25mg/kg dose group (5 females, 6 males), plus 10 normal controls (5 females and 5 males), 7 groups (δΐ), for intragastric administration Dosing for 6 weeks. During the test, one patient died of intragastric administration of tracheal intubation (female Qinggan Huayu 3.6g powder/kg), and 7 rats died of modeling, of which: 2 model control groups (female 1 (male, 1 male), Qinggan Huayu 0.9g powder / kg dose group 2 (female and male), Qinggan Huayu 1.8g powder / kg dose group 1 (female), compound armor Soft liver tablets 2g powder / kg dose group 1 (female), colchicum Alkali 0.25 mg / kg dose group 1 (; male;).

4.2 Determination of liver and spleen organ coefficient

After the last administration, the rats were fasted for 15 hours, the body weight was weighed, the liver and spleen were taken out, and the wet weight was weighed. The liver weight and spleen weight of 100 g body weight were used as the organ coefficient, and the difference between the dose groups was compared. 1.

Table 1 Effect of Qinggan Huayu Capsule on Organ Coefficient of Rats with Immunological Injury Hepatic Fibrosis (^士 SD) Dose (g Drug Number of Animals

Group liver coefficient spleen coefficient

Powder /kg) ' (only)

Normal control group 10 2.776±0.242 0.253±0.035 Model control group 10. 3.744±0.631## 0.366±0.037## Qinggan Huazao 嚢0.9 10 3.348±0.406 0.318±0.069 Qingganhua gelatin 嚢1.8 11 3.224±0.370* 0.321v0.057* Qinggan Huaji Capsule 3.6 11 3.187±0.303* 0.309±0.046** Compound 鳖曱soft liver tablets 2.0 11 3.645±0.491 0.327±0.047* colchicine 0.25mg 10 3.217±0.414* 0.317±0.050 * p<0. 01 (compared with the normal control group) p<0. 05 **p <0. 01 (compared with the model control group)

The results in Table 1 show that compared with the normal control group, the liver and spleen organ coefficients of the model control group were significantly increased; compared with the model control group, the liver and spleen of the liver phlegm and sputum sputum 1.8, 3.6 g powder / kg dose group The organ coefficient was significantly reduced, and the positive drug also had a similar effect.

4.3 Determination of blood biochemical indicators

For fasting rats, intraperitoneal anesthesia with pentobarbital sodium, blood was taken from the abdominal aorta, and sucked after centrifugation. Take the supernatant and measure ALT, AST, TP, ALB, G, A/G on the Olympus Au640 automatic biochemical analyzer. According to the instructions of the sialic acid (SA) assay kit produced by Nanjing Institute of Bioengineering, serum The sialic acid content was compared for each dose group, and the results are shown in Table 2. Table 2 Effect of Qinggan Huayu Capsule on Blood Biochemical Parameters in Rats with Immunological Injury Hepatic Fibrosis (±SD, n) Dose

ALT AST TP ALB G SA

Group (g powder A/G

(U/L) (U/L) (g L) (g L) (g/L) (mmol/L)

/kg) Normal control 34. ±7.22 105.3⁄428.9 59.QB.39 30.2t2.65 28.Sil.58 1.05 times 7 3. Gang.38 Model control 37.1±8.82 123.General.4 58.6B.24 29.QH2.85 29.6±2.17 0.93⁄40.13 3.79i0.47# Qinggan Huayu Capsule 0.9 56.9±39.1 155.3±57.2 59.3±3.19 29.4±1.80 29.8±2.69 0.99±0.12 3.63±0.41 Qinggan Huayu Capsule 1.8 0.2±13.7 107.8±54.9 60.7±2. 4 30.3±1.80 30.3±2.94 1.01±0.14 3.62±0.69 Qinggan Huayujiao 3.6 34.7±12.2 109.&B3.9 59.3J2.30 29.9il.79 2.66B0. 0 1.03±0.13 3. OM). 0 compound armor sheet 2.0 4.5±19.8 124. ± 3.1 59.8±3.06 29.&t2.14 30.OH2.03 1 ship 09 3. 6±0.50 colchicine 0.25mg 45 ±22.3 1262B5.9 612i2.71 30.6tl.92 30.6t2.97 1.01±0.14 3.55±0.68

#: p<0.05 (compared with normal control group) *: p<0.05 (compared with model control group) n: number of animals, same as table 1.

The results in Table 2 indicate that compared with the normal control group, the serum sialic acid content of the model control group was significantly increased, and the other indicators were not significantly different. Compared with the model control group, the serum sialic acid content of Qinggan Huayujiao 3.6g powder/kg dose group was significantly lower, and there was no significant difference among the other indicators.

4.4 Determination of liver hydroxyproline content

' Take about lg of liver expansion and weaving, add 9 times of normal saline for homogenization, even 3500 rpm / separation The heart was taken for 10 minutes. The supernatant was taken according to the instructions of the hydroxyproline kit produced by Nanjing Institute of Bioengineering. The hydroxyproline content in the liver was measured and the differences between the groups were compared. The results are shown in Table 3.

Table 3 Effect of Qinggan Huatan Capsule on Hepatic Hydroxyproline Content in Rats with Immunological Injury Hepatic Fibrosis (±SD) Dose Number of Animals Hydroxyproline

Group

(g powder/kg) (only) (ig/mgprot) normal control group 10 0.609±0.151 model control group 10 0.903±0.121## Qinggan Huaqi 嚢 0. 9 10 0.767±0.190 Qinggan Huayu 嚢1 8 11 0.726±0.163* 清肝化痴胶嚢 3. 6 11 Ο.όδθΐΟ.ΠΙ** Compound Biejia Ruangan Tablet 2 11 0.732±0.153* colchicine 0. 25mg 10 0.727±0.158* ρ<0.001 (Compared with normal control group) p<0.05 **p<0.01 (compared with model control group)

The results in Table 3 show that: Compared with the normal control group, the liver hydroxyproline content of the model control group was significantly increased, indicating that liver fibrosis had formed. Compared with the model control group, Qinggan Huayujiao 1.8, 3.6g powder/kg dose group significantly reduced the hydroxyproline content, and with the increase of dose, the effect was enhanced, indicating that the liver fibrosis has obvious prevention and treatment, positive control The medicinal compound Biejia Ruangan tablets and colchicine also have similar effects.

4.5 Histopathological examination

The liver was taken and fixed with 10% formalin. After routine sampling, alcohol gradient dehydration, Fisher Model 266Mp automatic paraffin embedding machine, Leica RM2135 slicing mechanism, HE, Masson two staining, Olympus coagulation: histopathological observation and photography were performed under the microscope according to the following grading standards, and the differences in each dose group were compared.

Grading criteria: References, the degree of liver fibrosis is divided into four levels:

"-,,: normal liver tissue, no fibrotic tissue hyperplasia

"+": There is fibrous tissue hyperplasia in the portal area and sinus.

"++": The structure of the hepatic lobule is destructive, the fibrous tissue proliferates to form a fibrous septum, and a small number of pseudolobules form a tendency.

"+++,,: Hepatic lobular structure destruction, fibrous tissue hyperplasia, ^ pseudolobules are widely formed. The results are shown in Table 4, Table 5. Table 4 Qinggan Huatan 嚢嚢 on liver fibers of rats with immune-damaging liver fibrosis Effect of degree of chemotherapy (HE staining) number of animals, HE staining, degree of liver fibrosis

Group P

(g raw drug/kg) (only) - + ++ +++ Normal control group 10 10 0 0 0 Model control group 10 1 3 1 5 <0.01 Qinggan Huayu gum 嚢 0. 9 10 3 1 1 5 >0.05 Qinggan Huayu Quyu 1. 8 11 7 1 1 2 <0.05 Qinggan Huayu Gelatin 3. 6 11 8 1 0 2 <0.05 Fufang Qigangan Tablet 2 11 5 2 3 1 <0.05 Colchicine 0 25mg 10 4 3 2 1 <0.05 Note: The number of different degrees of liver fibrosis is the number of animals in each dose group to achieve this level, Table 5 is the same as Table 4 Table 5 Effect of Qinggan Huayu Capsule on Hepatic Fibrosis in Rats with Immunological Injury Hepatic Fibrosis (Masson Staining)

Masson staining, liver fibrosis

Number of animals

Group degree P

(g raw medicine / kg) (only)

- + ++ +++ Normal control group 10 10 0 0 0 Model control group 10 1 3 1 5 <0.01 Qinggan Huaji capsule 嚢 0. 9 10 3 1 1 5 >0.05 Qinggan Huayu gum 嚢 1. 8 11 7 ' 1 1 2 <0.05 Qinggan Huaji Jiaojiao 3. 6 11 8 1 0 2 <0.05 Fufang Qigangan tablets 2 11 5 2 3 1 <0.05 Colchicine 0. 25mg 10 4 3 2 1 < 0.05

Table 4 and Table 5 show that compared with the normal control group, the degree of liver fibrosis in the model control group was significantly increased, indicating successful modeling. Compared with the model control group, the degree of liver fibrosis was significantly reduced in the 1.8 and 3.6 g powder/kg dose groups, indicating that the liver fibrosis has obvious preventive and therapeutic effects, and the effect is enhanced with increasing dose. The positive control compound compound 鳖曱 soft liver tablets and colchicine also have a significant effect.

5 test conclusions

The model of immunologically injured liver fibrosis was replicated by intravenous bovine serum albumin. The results showed that compared with the normal control group, the liver hydroxyproline content of the model control group was significantly increased, and the hyperplastic fibrous tissue divided the liver tissue into A wide range of pseudo-lobes; Qinggan Huayu gum 嚢 1.8, 3.6g powder / kg dose group liver hydroxyproline content was significantly reduced, liver fibrosis degree was significantly reduced, and with increasing dose, the effect increased, indicating Qinggan Huayu嚢 has obvious preventive and therapeutic effects on immune liver fibrosis. Experiment 2: Prevention and treatment of acute and chronic liver injury by Qinggan Huayu drug (pharmaceutical composition for treating liver fibrosis)

1. Experimental materials:

Qinggan Huayu Capsule: The hard capsule prepared according to Example 2, that is, the liver fibrosis drug of the present invention, the content is a reddish brown powder.

Biphenyl diester, produced by Peking Union Medical Co., Ltd., batch number 970419.

Animals: Kunming mice and SD rats were provided by the Experimental Animal Center of Hebei Province. The certificates were 04056 and 04057 respectively. During the observation period, the solid feed provided by the center was used for free drinking.

2. Test-risk method

2.1 Effects on acute liver injury induced by carbon tetrachloride in mice

METHODS: Sixty mice, half male and half female, weighing 20-25 g, were randomly divided into 6 groups, 10 in each group. The blank control group and the model control group were given water; Qinggan Huayu gum was divided into three dose groups: 0.9g, 1.8g and 3.6g/kg; biphenyldicarboxylate 0.2g/kg. All the above groups were administered by intragastric administration, and the volume was 0.2 ml/10 g body weight once a day for 7 consecutive days. After the sixth day of administration, 1% of the carbon tetrachloride oil solution was administered intraperitoneally, except for the blank control group, O.lml/lOg body weight.

One hour after the last administration (20 hours after carbon tetrachloride), the mice in each group were bled with blood, and the serum was separated, and the alanine aminotransferase (ALT) was determined by the Lai method.

Then each animal was dissected, taking a small piece of liver tissue from the same leaf and the same part, fixed in 10% formalin solution, and 4 pathological sections for histological examination, according to the following criteria (Liu Ting, etc., new Chinese medicine and Clinical Pharmacology 1999; 10 (4): 213-215) Evaluation of liver damage:

One: normal liver cells

+: Hepatocyte degeneration or punctate necrosis

++: Hepatocyte degeneration or focal necrosis, or diseased hepatocytes are less than 1/3 of hepatic lobules +++: Hepatic lobules 1/3-2/3 hepatocyte necrosis

++++: Liver lobules exceed 2/3 hepatocyte necrosis

The difference in ALT (t test) and the degree of liver injury (Ridit test) between the blank control group and the model control group and the model control group and each of the drug-administered groups were observed.

Results: The ALT of the model control group was significantly higher than that of the blank control group (PO.01), indicating that the model was successful. Qinggan Huayu Capsule 1.8g/kg significantly reduced ALT (PO.05), and bifendate had a significant reduction in ALT (P<0.01). The experimental results are shown in Table 6.

Pathological examination showed that the hepatic lobules in the model group were 1/3-2/3 and even more than 2/3 hepatocyte necrosis. The hepatic lobular cell necrosis in the liver and 3.6 g/kg group was lighter than the model (P>0.05). Hepatocyte necrosis or degeneration in the biphenyl diester group was significantly less than that in the model control group (P<0.05). The experimental results are shown in Table 7. Table 6: Effect of Qinggan Huayu capsule on transaminase in mice with acute carbon tetrachloride injury model. Group dose (g/kg) Number of animals (only) ALT (±s) (u) Blank control one 10 48.8± 38.5

Model control 1 10 293.9± 9.6 Qinggan Huayujiao 0.9 10 292.0±12.9 Qinggan Huashi stocks 1.8 10 285.3± 7.0* Qinggan Huashi stocks 3.6 10 288.7±11.6

Biphenyl diester 0.2 10 225.1±72.5**

*P<0.05, **P<0.01 vs. model control Table 7: Effect of Qinggan Huayu Capsule on Pathological Changes of Mouse Carbon Tetrachloride Acute Liver Injury Model Group Dose Animals

P value

Number

( g/kg (only) - + ++ +++ ++++ <0.01 )

Blank control one 8 8

Model control one 10 9 1

Qinggan Huashi Capsule 0.9 10 1 9 >0.05 Qinggan Huayu Capsule 1.8 10 2 8 >0.05 Qinggan Huayu Capsule 3.6 10 2 8 >0.05 Biphenyldicarboxylate 0.2 10 5 5 <0.05

2.2 The effect of D-galactosamine on acute liver function damage in mice

METHODS: Sixty mice, half male and half female, weighing 20-25 g, were randomly divided into 6 groups, 10 in each group. The blank control group and the model control group were given water; Qinggan Huayu gum was divided into three dose groups: 0.9g, 1.8g and 3.6g/kg; biphenyldicarboxylate 0.2g/kg. All the above groups were administered by intragastric administration, and the volume was 0.2 ml/10 g body weight once a day for 10 consecutive days.

One hour after the last administration, each mouse was intraperitoneally injected with D-galactosamine 650 mg/kg except for the blank control group. After 20 hours, the mice were sacrificed by arterial bleeding, blood serum was collected, and alanine was determined by the Reid method. Amino converting enzyme (ALT) and aspartate amino converting enzyme (AST) content. The significance of the difference between the blank control group and each of the drug-administered groups and the model control group was compared by t test.

Results: The ALT and AST contents of the model group were significantly higher than those of the blank control group, indicating that the model was successful. The ALT levels in the 0.9g, 1.8g and 3.6g/kg groups of Qinggan Huayu Capsule were significantly lower than those in the model control group (PO.05); 1.8g/kg could significantly reduce the AST content (PO.05). Biphenyl diester 9

The ALT content of the -21- group was also significantly lower than that of the model control group (PO.05, Table 8).

Table 8: Effect of Qinggan Huayu Capsule on acute liver function damage of D-~^lactosamine in mice Group dose Number of animals ALT (x±s) AST(x±s) (u)

(g/kg) (only) (u)

Blank control one 10 55±17 355±51 model control one 10 233±53 605±196 Qinggan Huayu gelatin 0.9 10 188±40* 519 soil 52 Qingganhuaji gelatin 101.8 10 183±47* 462士58 * Qinggan Huaqi sac 3.6 10 180 soil 49* 473± 86

Biphenyl diester 0.2 10 190±24* 603± 69

*Ρ<0.05, **Ρ<0.01 vs. model control

2.3 Effects of tetrachloroethylene on human liver injury in rats

Methods: SD rats, 110 males, weighing 225-349 g, were randomly divided into 6 groups. Except for 10 blank control groups, 20 rats in each group. The blank control group and the model group were given water; Qinggan Huayu capsule was divided into three dose groups: 0.9g, 1.8g and 3.6g/kg; and the positive control drug biphenyldicarboxylate 0.1g/kg. All were administered by intragastric administration, and the volume was Iml/lOOg body weight. Once a day for 10 consecutive weeks. In addition to the empty control, subcutaneous injection of 40% carbon tetrachloride peanut oil solution 3 ml/kg twice a week for 10 weeks.

On the next day after the last administration, blood was taken, aspartic acid aminotransferase (AST) and alanine aminotransferase (ALT) were determined by the Reid method, and total protein (TP) and bromine were determined by biuret method. The green method was used to determine albumin (ALB) and the albumin/globulin ratio (A/G) was calculated. Then the animals were sacrificed, and a small piece of liver was taken to measure the content of hepatic hydroxyproline. Finally, the liver was fixed with furfural, embedded in paraffin, stained with Masson, and scored according to the following histological grading criteria (Wang Baoen et al.: Journal of Hepatology, 1993: 1: 69-72). Level 0: normal liver tissue

Grade I: Collagen fibers extend outward from the portal or central vein

Grade II: Collagen fibers are clearly extended, but not yet connected to each other around the liver lobules

Grade III: Collagen fibers extend and connect, wrapping the entire liver lobules

Grade IV: Collagen fibers surround the hepatic lobules, causing the destruction of normal hepatic lobule structures, pseudo-lobes, but large, square ^ _ leaflets

Grade V: The liver is covered with small rounded false leaflets, forming a large square and small rounded false leaflets each accounting for 50%. Grade VI: The liver is covered with small round pseudolobules, and there are coarse hyperplastic collagen fibers between the pseudolobes. Ridit test, other indicators were tested by t test, comparing the significance of each drug group and model control group.

Results: 1. Surviving rats and body weight: The first injection of 40% carbon tetrachloride 3ml/kg was performed by intraperitoneal injection. After 3 days, the rats died, and then changed to subcutaneous injection, and some deaths were still observed. The most deaths occurred, and the middle dose group died less. Body weight changes: The weight of the high-dose group was lighter than that of the blank control group (P<0.05), which was indistinguishable from the model control group. See Table 9.

Table 9. Effect of Qinggan Huayu Capsule on the survival and body weight of carbon tetrachloride in rats

Group doses Number of original animals 10 weeks later Moving original body weight 10 weeks later

(g/kg) (only) Number of objects (only) (g) Weight (g) Blank control one 10 10 281±28 358±38 Model control one 20 10 289±28 339±42 Qinggan Huatan 嚢 0.9 20 12 286±27 338±35 Qinggan Huayujiao 1.8 20 15 290±25 347±26 Qinggan Huayu 嚢 20 20 20 287±25 325±26 Biphenyl diester 0.1 20 13 285±31 331±31

'PO.05 compared with the blank control 2, the effect on the conversion enzyme: Qinggan Huayu gum 嚢 1.8g and 3.6g / kg can increase the AST of elevated carbon tetrachloride (P <0.05). 3.6g/kg significantly decreased ALT (P<0.05), and bifendate significantly decreased AST and ALT (P<0.01). The experimental results are shown in Table 10. Table 10. Effect of Qinggan Huatan Capsule on Serum Converting Enzymes in Rats with Carbon Tetrachloride Chronic Hepatic Injury Group Dose (g/kg) Number of Animals (only) ALT ( x ± s ) ( u ) AST ( x ± s) ( u ) blank control - 10 380 ± 55 44 ± 9 model control - 10 589 ± 49 138 ± 34 clear liver phlegm and blood stasis 嚢 0.9 12 545 ± 71 146 ± 43 Qinggan idiot 嚢 1.8 15 525 ± 74 * 149±52 Qinggan Huayu Quercus 3.6 13 530±47* 109±21* Biphenyldiester 0.1 13 506±49 66±12

*Ρ<0.05, **Ρ<0.01 vs. model control

3. Effect on serum protein content: Each dose of Qinggan Huatan capsule and bifendate had no significant effect on serum total protein (TP) content (P>0.05); 3.6g/kg significantly increased albumin content (P<0.01), the A/G ratio also increased significantly (P<0.05). Biphenyldiester had no effect on the ratio of ALB and A/G. The experimental results are shown in Table 11. 39

-24- Table 11. Effect of Qinggan Huayu Capsule on protein content of rat carbon tetrachloride chronic liver injury Group dose Number of animals TP {≠) ALB(g/l) A/G ratio

(g/kg) (only)

Blank control one 10 69.1±5.8 44.1±2.7 1.84±0.41 Model control one 20 69.6±7.8 42.3=1=2.6 1.61±0.46 Qinggan Huayu limb 嚢 0.9 12 62.8±8.4 41.9±4.2 2.59±1.68 Qinggan Huayu嚢1.8 15 64.9±9.6 44.7±4.3 2.33±1.50 Qinggan Huayu Capsule 3.6 13 69.3±6.2 47.5±2.2 2.40±0.91* Biphenyldiester 0.1 13 68.1±8.8 41.6±1.7 1.84±1.13

*P<0.05, **Ρ<0.01 vs. model control

4. Effect on the content of hydroxyproline in liver tissue: Qinggan Huatan 嚢 3.6g/kg can significantly reduce the hydroxyproline content of liver tissue increased by carbon tetrachloride (PO.05), biphenyl diester It can also significantly reduce hepatic hydroxyproline content (PO.05). The experimental results are shown in Table 12.

Table 12. Effect of Qinggan Huatan Capsule on Hepatic Hydroxyproline Content in Rats with Chronic Hepatic Injury to Carbon Tetrachloride. Group Dose (g/kg) Number of Animals (only) Hepatic hydroxyproline content (x±s ) ( xg/kg ) blank control one 10 34 ± 8**

Model control 1 10 66±18

Qinggan Huayujiao 0.9 12 58±16

Qinggan Huayu Capsule 1.8 15 56±17

Qinggan Huayu Capsule 3.6 13 52±13*

Biphenyl diester 0.2 13 50±12*

*P<0.05, **P<0.01 vs. model control 5. Pathological examination of liver tissue: In the model control group, the structure of the hepatic lobule was completely destroyed, and the large square and round pseudolobules formed about half each. The hepatic collagen fibers were observed in the 1.8g and 3.6g/kg groups of Qinggan Huayu Capsule. They had not been or had been wrapped around the entire liver lobules. Some collagen fibers wrapped around the hepatic lobules to destroy the normal hepatic lobule structure. Leaflet. The scores were scored according to the histological grading criteria and their significance was tested by Ridit. Compared with the model control, the blank control group, L8g and 3.6g/kg were statistically significant (PO.05 and 0.01), and the experimental results are shown in Table 13. Table 13. Effect of Qinggan Huayu Capsule on Liver Pathology Grading of Carbon Tetrachloride Injury in Rats

Group dose number of animals pathological grade P value

( g/kg ) (only) 0 I II III IV V VI

Blank control - ^ 10 10 <0.01 model control 1 2 2 6

Qinggan Huayujiao 0.9 12 1 7 4 >0.05 Qinggan Huayujiao 1.8 15 3 3 4 4 1 <0.05 Qinggan Huayu Quercus 3.6 13 1 2 4 5 1 <0.05 Biphenyldiester 0.1 13 1 2 6 4 >0.05 2.4 Conclusion: In the acute liver function damage of mice with carbon tetrachloride, Qinggan Huayu capsule 嚢1.8g/kg can reduce ALT (P<0.05), and slightly relieve liver tissue damage. Acute liver function damage in D-galactosamine mice, Qinggan Huayu capsules 0.9g, 1.8g and 3.6g/kg can reduce ALT (P<0.05); 1.8g/kg can decrease AST (P<0.05) . Chronic liver function damage to rats with carbon tetrachloride, Qinggan Huayu capsule 嚢1.8g and 3.6g/kg can reduce AST (P<0.05), 3.6g/kg can decrease ALT (P<0.05); Gelatin 嚢 3.6g / kg can increase serum ALB content (P <0.05) and A / G ratio (P <0.05); The hydroxyproline content in liver tissue was decreased (P<0.05). The liver pathological grade, Qinggan Huayu capsule 嚢1.8g and 3.6g/kg could significantly reduce the degree of lesion (P<0.05).

Claims

Rights request

A pharmaceutical composition for treating liver fibrosis, characterized in that it is mainly prepared from the following raw materials of weight: 10 to 30 parts of gentian, 20 to 40 parts of medlar (vinegar), 30 to 50 of jaundice Parts, three edges (vinegar) 15~35 parts, salvia miltiorrhiza 25~45 parts, atractylodes 15~35 parts, astragalus 10~30 parts.

2. The pharmaceutical composition according to claim 1, wherein the amount of each drug substance is: 20 parts of gentian, 30 parts of medlar (vinegar), 40 parts of astragalus, 25 parts of trigonal (vinegar), 35 parts of salvia miltiorrhiza 25 Atractylodes and 20 Astragalus.

The pharmaceutical composition according to claim 1 or 2, wherein the drug substance further comprises 10 - 30 parts of dried tangerine peel and 10 - 30 parts of Polygonum cuspidatum.

The pharmaceutical composition according to claim 3, wherein the amount of dried tangerine peel is 20 parts, and the amount of tiger stick is 20 parts.

The pharmaceutical composition according to claim 1 or 2, wherein the drug substance further contains 5 to 15 parts of licorice.

The pharmaceutical composition according to claim 5, wherein the licorice is used in an amount of 10 parts.

7. The method for preparing a pharmaceutical composition according to claim 1, comprising the steps of: a. extracting from sputum (vinegar), atractylodes water vapor distillation, collecting volatile oil, and preparing for use, and volatile oil may also be prepared by β-cyclodextrin inclusion. Using, the distilled aqueous solution is collected separately;

b. Astragalus, Sanling (vinegar), Astragalus, Dan are taken in water reflux extraction, the extracts are combined, filtered, and the filtrate is combined with the aqueous solution obtained in step a, concentrated, ethanol added, refrigerated overnight, filtered, supernatant Standby

c gentian and ethanol are heated to reflux, the extract is filtered, and the filtrate is combined with the supernatant obtained in step b, and the ethanol is recovered to an alcohol-free taste and concentrated;

d. Combine the β-cyclodextrin inclusion compound of the volatile oil or volatile oil obtained in the step a and the concentrate obtained in the step c.

8. The method of claim 7 wherein the tangerine peel is added in step a and the knotweed is added in step b.

9. The method of claim 8 wherein licorice is added in step b.

The method according to claim 7, wherein the composition obtained in the step d and various pharmaceutical excipients are prepared into any oral dosage form by a conventional Chinese medicine preparation method.