CN108126134B - A kind of traditional Chinese medicine compound composition for treating liver injury, preparation method and application thereof - Google Patents

Abstract

The invention discloses a traditional Chinese medicine compound composition for treating liver injury, which is prepared from the following raw materials in parts by weight: 20-30 parts of purslane, 15-30 parts of cortex albiziae, 15-20 parts of raw malt, 5-10 parts of honey-fried licorice root, 10-20 parts of perilla stem and 15-20 parts of poria cocos. In addition, the invention also provides a preparation method and application of the traditional Chinese medicine compound composition for treating liver injury. The traditional Chinese medicine composition disclosed by the invention is simple in composition and reasonable in proportion, and can obviously relieve symptoms such as inappetence, tiredness, hypodynamia, abdominal distension, eructation, nausea, vomiting, weight loss, fullness and dull pain in the liver area or right upper abdomen and the like during liver injury, and the accumulation of liver fat is obviously reduced. The traditional Chinese medicine composition has good curative effect, no side effect and convenient taking after being taken for a long time, and provides a good choice for the patients with liver injury.

Description

A kind of traditional Chinese medicine compound composition for treating liver injury, preparation method and application thereof

technical field

The invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine compound composition for treating liver injury, a preparation method and an application thereof.

Background technique

Liver injury is one of the most important common diseases affecting human health. Various harmful factors such as viruses, biology, drugs, physicochemicals, and alcohol can cause liver damage. The etiology of liver injury diseases is complex, and its mechanism is still unclear, but it is currently believed to mainly involve organelle damage, changes in cell structure, lipid peroxidation, lipodystrophy, changes in enzyme activity, and disruption of ion balance. Among them, lipid metabolism disorders can directly lead to hepatic steatosis and cause damage. At present, it is believed that it involves at least the assembly damage of triglyceride-apolipoprotein complex, the damage of membrane transport function, the increase of lipid storage and the damage of mitochondrial oxidation function.

The prevention and treatment of liver injury diseases by drugs has become an important topic of social and medical attention. Although the development of drugs for the treatment of liver injury diseases is still in progress, the application of existing chemical drugs such as interferon and arabinoside is limited due to their high price, difficulty in popularization and many adverse reactions.

Traditional Chinese medicine has a long history in the treatment of liver injury diseases. According to the clinical manifestations of liver injury, in traditional Chinese medicine, it mostly belongs to the categories of diseases such as "flank pain", "jaundice", "accumulation" and "bloating". The causes are often exogenous damp heat, epidemic diseases Qi; improper diet, overeating fat and sweet; excessive drinking; drug toxicity and liver toxins in industry and the environment, etc. act on the human body. Therefore, the evil of dampness and poison is the main cause of this disease, and the deficiency of liver stagnation and spleen, and the evil love of positive deficiency are the pathological basis of this disease. According to the common pathogenesis of liver injury, the principles of clearing away heat and removing dampness and detoxification, soothing the liver and resolving stagnation, promoting blood circulation and removing blood stasis, strengthening the spleen and removing phlegm are often used, but the curative effect needs to be further improved. Therefore, under the guidance of the theory of traditional Chinese medicine, it is necessary to select a traditional Chinese medicine compound composition of natural medicine with good liver protection effect as a treatment method for patients with liver injury.

SUMMARY OF THE INVENTION

The invention provides a traditional Chinese medicine compound composition for treating liver injury, which solves the problems in the prior art that the chemical drugs for treating liver injury are expensive, difficult to popularize and have many adverse reactions, and also solves the problems that the traditional Chinese medicine for treating liver injury has poor curative effect. ideal question.

The first object of the present invention is to provide a traditional Chinese medicine compound composition for the treatment of liver injury. 15-20 servings, 5-10 servings of licorice root, 10-20 servings of Perilla stem, 15-20 servings of Poria.

Preferably, the raw materials for the preparation of the traditional Chinese medicine compound composition for treating liver injury are composed of the following components in parts by weight: 30 parts of purslane, 30 parts of Albizia Julibrissin bark, 20 parts of raw malt, 10 parts of licorice root, basil stems 20, Poria 20.

The second object of the present invention is to provide a preparation method of a traditional Chinese medicine compound composition for the treatment of liver injury, which is specifically implemented according to the following steps:

Step 1, take by weight 20-30 parts of purslane, 15-30 parts of Albizia Julibrissin bark, 15-20 parts of raw malt, 5-10 parts of licorice root, 10-20 parts of perilla stem, 15-20 parts of Poria cocos ,spare;

Step 2, pulverize the purslane weighed in step 1 and pass through a 30-mesh sieve to obtain purslane coarse powder, add petroleum ether equivalent to 10 times the weight of the purslane coarse powder to the purslane coarse powder for reflux extraction Degreasing for 1 h, and concentrating under reduced pressure after refluxing to obtain degreasing purslane coarse powder;

Add 80% ethanol solution equivalent to 10 times the weight of the degreasing purslane coarse powder to the defatted purslane coarse powder and soak it for 0.5h. After soaking, reflux extraction for 2 times. Filter, combine the filtrates twice, and concentrate under reduced pressure until there is no ethanol smell to obtain a concentrated solution;

Add distilled water to the concentrated solution, and disperse the concentrated solution into a medicinal solution with a crude drug content of 0.3 g/ml; put the medicinal solution on AB-8 macroporous adsorption resin, first eluate with 2 times the column volume of distilled water, and discard water eluent; then eluted with 4 times the column volume, the mass concentration of 10% ethanol solution, discard the first alcohol eluent; and then use 6 times the column volume, the mass concentration of 80% ethanol solution elution, collecting the second alcohol eluent, and concentrating the second alcohol eluent under reduced pressure, drying and pulverizing to obtain a purslane extract;

Step 3, mix the acacia peel, raw malt, licorice, basil stalk, and Poria that were weighed in step 1, and then add the total weight equivalent to the acacia peel, raw malt, licorice, basil stalk, and Poria. Soak 5 times of water for 30min, decoct for 1h after soaking, filter to obtain the first filtrate and filter residue; add water equivalent to 3 times the weight of the filter residue to the filter residue, decoct for 0.5h, filter to obtain the second filtrate, Combine the first filtrate and the second filtrate, mix well, and concentrate to a relative density of 1.2-1.4 to obtain a mixed extract;

In step 4, the purslane extract in step 2 and the mixed extract in step 3 are evenly mixed to obtain the traditional Chinese medicine compound composition for treating liver injury.

The third object of the present invention is to provide an application of a traditional Chinese medicine compound composition for treating liver injury in the preparation of a pharmaceutical preparation for treating liver injury.

Preferably, the traditional Chinese medicine compound composition for treating liver injury is prepared into a pharmaceutically acceptable pharmaceutical preparation according to a conventional method.

Preferably, the pharmaceutical dosage form of the pharmaceutical preparation is oral liquid, capsule, granule, pill, powder, microcapsule or drop pill.

Pharmacological action and efficacy of each raw material traditional Chinese medicine in the present invention:

1. Portulaca oleracea: cold in nature, sweet and sour in taste; enters the heart, liver, spleen, and large intestine meridians. Clearing away heat and detoxifying, removing dampness with water, dispersing blood and reducing swelling, reducing inflammation and relieving pain, hemostasis and cooling blood. Indications of dysentery, enteritis, nephritis, postpartum uterine bleeding, blood in the stool, mastitis and other diseases.

2. Acacia skin: sweet, flat. Return to the heart, liver and lung meridians. Relieve depression and soothe the nerves, activate blood and reduce swelling. For restlessness, depression and insomnia, pulmonary abscesses and swollen, falling and fluttering pain.

3, raw malt: sweet, flat. Returning to the spleen and stomach meridians, it has the effect of soothing the liver, promoting qi and relieving stagnation. It is used for flank pain, abdominal pain, etc. due to stagnation of liver qi or disharmony between the liver and stomach. Zhang Xichun, a famous doctor in modern times, is good at using raw malt to soothe the liver and raise the qi of the liver. Zhang Xichun pointed out that “although malt is a medicine for the spleen and stomach, it is really good at soothing the liver qi (soothing the liver is suitable for raw use, but it is ineffective when used in frying). Covering the liver is spring when it is used, and wood is used for the five elements. The liver seeks the same qi, so it is good to relieve it. The liver is mainly responsible for dredging the qi for the kidney, and its power can relieve the liver, and it is good to help the liver wood to dredge the qi to promote the kidney qi.”

4, Zhigancao: Qi medicine. Sweet and flat. Can invigorate the spleen and replenish qi, relieve cough and phlegm, clear away heat and detoxify, relieve acute pain, and mix various medicines. For weak spleen and stomach, insufficiency of qi, cough and asthma, carbuncle sore, abdominal spasm and acute pain, etc.

5. Perilla terrier: Xin, warm. Return to the lung and spleen meridians. Perilla is Xin Wen Xing San, the leaves are as light as the lungs, can disperse wind-cold, disperse the lungs and relieve cough; stems are like the spleen and stomach, good at promoting qi and neutralization, regulating qi and relieving miscarriage.

6. Poria: sweet, light, and flat, it has the effects of diuretic dampness, invigorating the spleen and stomach, calming the heart and soothing the nerves. . "Compendium of Materia Medica": Poria's smell is light and infiltrating, its nature is upward, it produces body fluids, opens the stomach, nourishes water sources and descends, and facilitates urination. Therefore, Zhang Jiegu said that it belongs to Yang, floats and rises, and its nature is also said; Dongyuan It is called the yin within the yang, descending and descending, and it is said that its power is also.

The present invention selects purslane as the monarch medicine, which has the functions of clearing away heat and detoxifying, nourishing the liver and dredging collaterals, dispelling dampness and improving the stomach. The acid enters the liver, nourishes the yin of the liver and nourishes the liver body, and the power of purslane to dispel dampness and stomach helps to transport and transform water and grains to generate qi and blood. The minister uses acacia peel, raw malt, and roasted licorice. The acacia peel is sweet and light and flat in nature, which has the effect of strengthening the spleen and infiltrating dampness. Promote qi and eliminate food, help the spleen and stomach to transport and transform, soothe the liver and relieve stagnation, and regulate the liver qi; licorice and licorice, enter the spleen and stomach meridians, have the effect of protecting the spleen and stomach, clearing away heat and producing body fluid, and have the dual effect of nourishing and eliminating pathogens for those with deficiency of stomach and body fluid. . Accompanied by perilla stems and tuckahoe, the perilla stems are pungent and warm, and the qi is neutralized, and the qi is regulated and the fetus is relieved. All medicines are used together to nourish the liver and spleen, detoxify and infiltrate dampness.

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

The traditional Chinese medicine compound composition of the present invention is used for treating liver injury, regulating and reducing the accumulation of fat in the liver. And based on the theory of traditional Chinese medicine, the combination of syndrome differentiation and disease differentiation, in terms of drug use, it is relatively concentrated on the functions of soothing the liver, regulating qi and strengthening the spleen, reducing phlegm and dampness and reducing turbidity, promoting blood circulation and removing stasis, eliminating accumulation and guiding stagnation. , Removing dampness and reducing turbidity, strengthening the spleen and nourishing the kidney, promoting blood circulation and removing blood stasis.

The traditional Chinese medicine composition of the present invention is simple in composition and reasonable in proportion, and has been proved by experiments to be effective for inappetence, fatigue, abdominal distension, belching, nausea, vomiting, weight loss, fullness and dull pain in the liver area or upper right abdomen, etc. Symptoms were significantly relieved, and liver fat accumulation was significantly reduced. The traditional Chinese medicine composition of the invention is taken for a long time, has good curative effect, has no side effects, is convenient to take, and provides a good choice for the majority of patients with liver injury.

Detailed ways

In order to enable those skilled in the art to better understand that the technical solutions of the present invention can be implemented, the present invention will be further described below with reference to specific embodiments, but the embodiments are not intended to limit the present invention.

The purslane, acacia bark, raw malt, licorice root, perilla stem, and Poria used in the embodiments of the present invention are all commercially available, and the test methods are conventional methods unless otherwise specified.

Example 1

A traditional Chinese medicine compound composition for treating liver injury, the preparation raw material is composed of the following components in parts by weight: 30 parts of purslane, 30 parts of Albizia japonicus bark, 20 parts of raw malt, 10 parts of licorice root, and 20 parts of perilla stems , Poria 20 copies.

Specifically, follow the steps below:

Step 1, take by weight 30 parts of purslane, 30 parts of Albizia Julibrissin bark, 20 parts of raw malt, 10 parts of licorice root, 20 parts of perilla stems, 20 parts of Poria, for subsequent use;

Step 2, pulverize the purslane weighed in step 1 and pass through a 30-mesh sieve to obtain purslane coarse powder, add petroleum ether equivalent to 10 times the weight of the purslane coarse powder to the purslane coarse powder for reflux extraction Degreasing for 1 h, and concentrating under reduced pressure after refluxing to obtain degreasing purslane coarse powder;

Add 80% ethanol solution equivalent to 10 times the weight of the degreasing purslane coarse powder to the defatted purslane coarse powder and soak it for 0.5h. After soaking, reflux extraction for 2 times. Filter, combine the filtrates twice, and concentrate under reduced pressure until there is no ethanol smell to obtain a concentrated solution;

Add distilled water to the concentrated solution, and disperse the concentrated solution into a medicinal solution with a crude drug content of 0.3 g/ml; put the medicinal solution on AB-8 macroporous adsorption resin, first eluate with 2 times the column volume of distilled water, and discard water eluent; then eluted with 4 times the column volume, the mass concentration of 10% ethanol solution, discard the first alcohol eluent; and then use 6 times the column volume, the mass concentration of 80% ethanol solution elution, collecting the second alcohol eluent, and concentrating the second alcohol eluent under reduced pressure, drying and pulverizing to obtain a purslane extract;

Step 3, mix the acacia peel, raw malt, licorice, basil stalk, and Poria that were weighed in step 1, and then add the total weight equivalent to the acacia peel, raw malt, licorice, basil stalk, and Poria. Soak 5 times of water for 30min, decoct for 1h after soaking, filter to obtain the first filtrate and filter residue; add water equivalent to 3 times the weight of the filter residue to the filter residue, decoct for 0.5h, filter to obtain the second filtrate, Combine the first filtrate and the second filtrate, mix well, and concentrate to a relative density of 1.2-1.4 to obtain a mixed extract;

In step 4, the purslane extract in step 2 and the mixed extract in step 3 are evenly mixed to obtain a traditional Chinese medicine compound composition for treating liver injury;

After crushing the above-mentioned traditional Chinese medicine compound composition, add appropriate amount of auxiliary materials, mix uniformly, granulate, sieve, and dry to prepare a granular type traditional Chinese medicine compound composition for treating liver injury.

Example 2

A traditional Chinese medicine compound composition for treating liver injury, the preparation raw material is composed of the following components in parts by weight: 20 parts of purslane, 15 parts of Albizia Julibrissin bark, 15 parts of raw malt, 5 parts of licorice root, and 10 parts of perilla stems , 15 parts of Poria.

The preparation method of Example 2 is the same as that of Example 1, except that the formula in Example 1 is replaced with that of Example 2.

The traditional Chinese medicine compound composition prepared in Example 2 is pulverized and then added with appropriate amount of auxiliary materials, mixed, sieved, granulated, and dried to obtain the traditional Chinese medicine compound composition for treating liver injury in pill form.

Example 3

A traditional Chinese medicine compound composition for treating liver injury, the preparation raw material is composed of the following components in parts by weight: 25 parts of purslane, 20 parts of Albizia Julibrissin bark, 18 parts of raw malt, 8 parts of licorice root, 15 parts of perilla stem , 15 parts of Poria.

The preparation method of Example 3 is the same as that of Example 1, except that the formula in Example 1 is replaced with that of Example 3.

The traditional Chinese medicine compound composition prepared in Example 3 is pulverized and then added with appropriate amount of auxiliary materials, mixed, granulated, sieved, and dried. Injured traditional Chinese medicine compound composition.

Examples 1-3 have all prepared the traditional Chinese medicine compound composition for the treatment of liver injury with good effect. Since the effects of the traditional Chinese medicine compound composition for the treatment of liver injury prepared in Examples 1-3 are basically parallel, only the implementation of The traditional Chinese medicine composition prepared in Example 1 is used to illustrate the effect of the present invention.

The effects of the present invention are further described below using animal tests.

1. Effects on the mouse model of acute alcoholic liver injury

1. Experimental materials

1.1 Experimental drugs

The traditional Chinese medicine compound combination for the treatment of liver injury prepared in embodiment 1;

Bifendiester Dropping Pills, batch number: A020141206, Wanbangde Pharmaceutical Group Co., Ltd.

1.2 Experimental reagents

Ethanol, Tianjin Zhiyuan Chemical Reagent Co., Ltd., batch number 2015122012; alanine aminotransferase (ALT) kit, batch number: 1507053; aspartate aminotransferase (AST) kit, batch number: 1507073; the above drugs are all provided by Shanghai Provided by Fosun Long March Medical Science Co., Ltd.

Coomassie brilliant blue protein kit, batch number: 20160118; superoxide dismutase (SOD) kit, batch number: 20160121; malondialdehyde (MDA) kit, batch number: 20160119; the above drugs are provided by Nanjing Jiancheng Bioengineering Institute .

1.3 Experimental Instruments

AU400 Automatic Biochemical Analyzer, Olympus, Japan; Cgtation3 CYT3 MFUDG Microplate Reader, Berton Instrument Co., Ltd., USA; UV-2000 UV-Vis Spectrophotometer, Unico (Shanghai) Instrument Co., Ltd.; TGL-16G Desktop centrifuge, Shanghai Anting Scientific Instrument Factory; electronic balance, METTLER TOLEDO Instruments (Shanghai) Co., Ltd.; DMI4000B intelligent inverted fluorescence microscope, Leica Instrument Co., Ltd., Germany.

1.4 Experimental animals

Kunming mice, SPF grade, half male and half female, weighing 18-22g, provided by Jinan Pengyue Laboratory Animal Breeding Co., Ltd., production license number: SCXK (Lu) 20140007, experimental animal use license number: SYXK (Yu) 2015-0005.

2. Experimental method

2.1 Preparation and administration of acute alcoholic liver injury model

60 KM mice were randomly and evenly divided into 6 groups, namely normal control group, model group, low, medium and high dose groups of traditional Chinese medicine compound composition (12.58, 25.16, 50.32g/kg) and bifendate group ( 5.625 mg/kg). 10 in each group, half male and half female. The traditional Chinese medicine compound composition administration group and the bifendate group were respectively given different doses of the traditional Chinese medicine compound composition and bifendate by gavage at a volume of 20ml/kg, and the normal control group and the model group were given the same volume of distilled water every day. Administered once a day, continuous administration for 7d. One hour after the last administration, except for the normal control group, the other groups were given 50% ethanol 14ml/kg by gavage to establish the acute alcoholic liver injury model.

2.2 Measurement indicators

After ethanol gavage, the rats were fasted and watered. After 12 hours, the eyeballs were removed and blood was collected. The serum was separated, and the levels of serum liver function indexes ALT and AST were determined by automatic biochemical analyzer. The right lobe of liver was collected by adding normal saline to make 10% liver homogenate, and the supernatant was collected by centrifugation. The activity of SOD in liver tissue homogenate was detected by microplate method, and the content of MDA was detected by colorimetric method. After the serum was collected, the liver was excised, and the left lobe of the liver was taken and fixed in 10% formaldehyde solution for histopathological examination. The liver tissue was fixed in 10% formaldehyde solution, embedded in paraffin and sliced, stained with HE, and histopathological changes were observed under light microscope.

Among them, the grading of alcoholic liver injury during the histopathological examination of the liver is as follows:

"-" Hepatocyte structure is complete, the boundary is clear, the hepatic cord is neatly arranged, and there is no edema, fatty degeneration and necrosis of the hepatocyte;

"+" hepatocytes showed mild edema, slightly increased cell volume, and a small amount of inflammatory cell infiltration in hepatocytes;

"++" hepatocytes showed moderate edema, disordered arrangement of hepatocyte cords, significant increase in the volume of lipid droplets in some cytoplasms, and infiltration of multiple inflammatory cells;

"+++" Hepatocytes showed severe edema, the volume of lipid droplets in some cytoplasms increased significantly, the structure of hepatic lobules was blurred, the infiltration of inflammatory cells was obvious, and hepatocyte necrosis occurred.

2.3 Statistical methods

”表示，两组间比较采用t检验，多组间比较采用单因素方差法，组间两两比较方差齐性时用LSD方法分析，方差不齐时用Dunnett’s方法分析。病理半定量分析采用 Radit法。SPSS 17.0 statistical software was used, and the measurement data were calculated as "mean ± standard deviation".

” indicates that the t test was used for the comparison between two groups, the one-way ANOVA method was used for the comparison between multiple groups, the LSD method was used for the comparison between the two groups when the variance was homogeneous, and the Dunnett’s method was used for the analysis when the variance was unequal. Pathological semi-quantitative analysis used Radit Law.

3. Experimental results

The effects of traditional Chinese medicine compound compositions on the liver function of mice with acute alcoholic liver injury are shown in Table 1-3.

(n＝10)Table 1 Effects on liver function in mice with acute alcoholic liver injury

(n=10)

Note: *P<0.05, **P<0.01 compared with the model group.

As can be seen from Table 1, compared with the normal control group, the serum ALT and AST values of the mice in the model group were significantly increased (P<0.01), indicating that the model of alcoholic liver injury in mice was successful. Compared with the model group, the ALT and AST values in the low-dose, medium-dose, and high-dose groups and the bifendate group were significantly decreased (P<0.01 or P<0.05).

(n＝10)Table 2 Effects on SOD and MDA in liver tissue of mice with acute liver injury

(n=10)

Note: *P<0.05, **P<0.01 compared with the model group.

As can be seen from Table 2, compared with the normal control group, the levels of SOD and GSH in the liver tissue of the mice in the model group were significantly decreased (P<0.01), and the content of MDA was significantly increased (P<0.01). Compared with the model group, the levels of SOD and GSH in the liver tissue of the mice in each dose group of the compound Chinese medicine composition were significantly increased, while the content of MDA was significantly decreased (P<0.01). certain protection.

Table 3 Histopathological examination results of mouse liver (n=10)

It can be seen from Table 3 that the hepatic lobules of the mice in the normal control group were normal in structure, with radial arrangement of hepatocyte cords, no edema, degeneration and necrosis of hepatocytes, and no inflammatory cell infiltration in the portal area. In the model group, the arrangement of the hepatic cords was disordered, the cytoplasm was loose, the edema of the hepatocytes increased, and there was obvious fatty degeneration. Compared with the model group, the degree of hepatocyte lesions in mice in each dose group of the compound Chinese medicine composition was significantly reduced, the lipid droplets in the cytoplasm were reduced, the steatosis was significantly reduced, and the edema and necrosis of hepatocytes were not obvious. The results showed that each dose group of the traditional Chinese medicine compound composition could inhibit the formation of hepatocyte necrosis and steatosis to different degrees, among which the high dose group was more obvious.

2. Influence on the model of drug-induced liver injury induced by paracetamol in mice

1 Experimental material

1.1 Experimental drugs

The traditional Chinese medicine compound combination for the treatment of liver injury prepared in embodiment 1;

Bifendiester Dropping Pills, batch number: A020141206, Wanbangde Pharmaceutical Group Co., Ltd.;

Acetaminophen sustained-release tablets, batch number: H20010394, Shanghai Johnson & Johnson Pharmaceutical Co., Ltd.

1.2 Experimental reagents

Alanine aminotransferase (ALT) kit, batch number: D1507053; Aspartate aminotransferase (AST) kit, batch number: D1507073; all provided by Shanghai Fosun Long March Medical Science Co., Ltd. Coomassie brilliant blue protein kit, batch number: 20160323; superoxide dismutase (SOD) kit, batch number: 20160322; malondialdehyde (MDA) kit, batch number: 20160311; all provided by Nanjing Jiancheng Bioengineering Institute.

1.3 Experimental Instruments

AU400 automatic biochemical analyzer, Olympus, Japan; Cytation3 CYT3 MFUDG microplate reader, Berton Instrument Co., Ltd., USA; UV-2000 UV-Vis Spectrophotometer, Unico (Shanghai) Instrument Co., Ltd.; TGL-16G Desktop centrifuge, Shanghai Anting Scientific Instrument Factory; AX3242H Electronic Balance, METTLER TOLEDO Instruments (Shanghai) Co., Ltd.; DMI4000B Intelligent Inverted Fluorescence Microscope, Leica Instrument Co., Ltd. Germany.

1.4 Experimental animals

Kunming mice, SPF grade, half male and half female, weighing 18-22g, provided by Jinan Pengyue Laboratory Animal Breeding Co., Ltd., production license number: SCXK (Lu) 20140007, experimental animal use license number: SYXK (Yu) 2015-0005.

2 Experimental methods

2.1 Modeling and processing

60 mice were taken and randomly divided into 6 groups, 10 mice in each group, half male and half female, respectively normal control group, paracetamol model group, bifendate group (5.625mg/kg), Chinese medicine compound composition low, medium, and High dose group (12.58, 25.16, 50.32g/kg). The traditional Chinese medicine compound composition administration group and the bifendate group were respectively given different doses of the traditional Chinese medicine compound composition and bifendate by gavage at a volume of 20ml/kg, and the normal control group and the model group were given the same volume of distilled water by gavage every day. Administered once a day, continuous administration for 7d. 6 hours after the last administration, except for the normal control group, paracetamol 0.2ml/10g was given by intragastric administration to cause acute liver injury, and the normal control group was given the same volume of distilled water by intragastric administration. The animals were fasted and watered, and the body weight was weighed 12 hours after the last administration, and the animals were treated. The eyeball was removed for blood, the serum was separated, and the liver was quickly removed. The right lobe of liver was taken to make 10% liver homogenate, and the supernatant was collected by centrifugation. The left lobe of liver was taken and fixed in 10% formaldehyde solution for histopathological examination.

Among them, the grading of alcoholic liver injury during the histopathological examination of the liver is as follows:

"-" Hepatocyte structure is normal, hepatocyte cord and hepatocytes are normal;

"+" The structure of hepatocytes is relatively normal, the arrangement of hepatocytes is more regular, the hepatocytes are mildly degenerated, and a small amount of inflammatory cell infiltration can be seen;

"++" Hepatocyte structural deformation, disordered arrangement of hepatocytes, moderate degeneration and necrosis of hepatocytes, and obvious infiltration of multiple inflammatory cells;

"+++" The structure of hepatocytes was significantly changed, the arrangement of hepatocytes was disordered, and large areas of degeneration and necrosis were seen, and the infiltration of inflammatory cells was obvious.

2.2 Measurement indicators

Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were determined by automatic biochemical analyzer. The activity of superoxide dismutase (SOD) in liver tissue homogenate was detected by microplate method, and the content of malondialdehyde (MDA) was detected by colorimetric method. HE staining, and histopathological changes were observed under light microscope.

2.3 Statistical methods

”表示，两组间比较采用t检验，多组间比较采用单因素方差法，组间两两比较方差齐性时用LSD方法分析，方差不齐时用Dunnett’s方法分析。病理半定量分析采用 Radit法。SPSS 17.0 statistical software was used, and the measurement data were calculated as "mean ± standard deviation".

” indicates that the t test was used for the comparison between two groups, the one-way ANOVA method was used for the comparison between multiple groups, the LSD method was used for the comparison between the two groups when the variance was homogeneous, and the Dunnett’s method was used for the analysis when the variance was unequal. Pathological semi-quantitative analysis used Radit Law.

3. Experimental results

Table 1-3 shows the effects of traditional Chinese medicine compound compositions on the liver function of mice with paracetamol-induced liver injury.

(n＝10)Table 4 Effects on serum ALT and AST in mice with paracetamol-induced liver injury

(n=10)

Note: *P<0.05, **P<0.01 compared with the model group.

As can be seen from Table 4, compared with the normal control group, the levels of ALT and AST in the serum of the mice in the model group were significantly increased (P<0.01), indicating that the modeling was successful. Compared with the model group, the levels of ALT and AST in each dose group of bifendate and the traditional Chinese medicine compound composition can be reduced (P<0.01), and the high-dose group of the traditional Chinese medicine compound composition has the most obvious effect.

(n＝10)Table 5 Effects on SOD and MDA in liver tissue of mice with paracetamol-induced liver injury

(n=10)

Note: *P<0.05, **P<0.01 compared with the model group.

It can be seen from Table 5 that compared with the normal control group, the content of MDA in the liver tissue of the mice in the model group was significantly increased (P<0.01), and the activity of SOD was significantly decreased (P<0.01). Compared with the model group, each dose group of bifendate and traditional Chinese medicine compound composition could reduce the content of MDA and increase the activity of SOD (P<0.01 or P<0.05).

Table 6 The grades of liver tissue lesions in mice in each group (n=10)

It can be seen from Table 6 that compared with the model group, there were significant statistical differences between each dose group of the compound Chinese medicine composition and the bifendate group (P<0.01 or P<0.05). The pathological examination of liver tissue showed that the structure of the hepatic lobules in the normal control group was normal, the liver cells were arranged regularly, the size of the liver cells was uniform, the nucleus was obvious, the cytoplasm was abundant, and the hepatocyte cords were arranged radially and neatly around the central vein. In the paracetamol model group, the arrangement of hepatocytes was disordered, the cords of hepatocytes disappeared, the edema and volume of hepatocytes increased, and the hepatocytes had different degrees of degeneration and necrosis, showing obvious inflammatory cell infiltration. This indicates that the paracetamol-induced mouse drug-induced liver injury model was successfully replicated. The degeneration and necrosis of hepatocytes in the administration group of the traditional Chinese medicine compound composition were mild, and the hepatic cord had different degrees of recovery. Among them, the effect of the high-dose group of the traditional Chinese medicine compound composition was the most obvious.

The above-mentioned ethanol-induced acute alcoholic liver injury model in mice and paracetamol-induced mouse liver injury model have confirmed that the traditional Chinese medicine compound composition of the present invention has a definite hepatoprotective effect, so it can be used for preparing medicines for treating liver injury.

On the basis of ensuring safety in animal experiments, the Chinese medicine compound composition for treating liver injury prepared in Example 1 of the present invention was clinically applied, and good technical effects were obtained through clinical application of 46 patients with liver injury. The patient took the above traditional Chinese medicine compound composition twice a day for 30 days, the clinical symptoms disappeared or basically disappeared or the symptoms improved, the liver function was normal or significantly recovered, or the values of alanine aminotransferase and aspartate aminotransferase decreased by more than 50%, and the patients reached 44. people, the effective rate reaches 95.7%. It can be seen from the clinical test that the traditional Chinese medicine compound composition of the present invention has the effect of treating liver injury, is effectively used for preparing medicines for treating liver injury, realizes the application of the traditional Chinese medicine compound composition in the treatment of liver injury, and has significant economic and social benefits

It should be noted that when the claims of the present invention relate to the numerical range, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. 3 is the same, in order to avoid redundant description, the present invention describes the preferred embodiments, but those skilled in the art can make additional changes and modifications to these embodiments once the basic inventive concept is known. Therefore, the appended claims are intended to be construed to include the preferred embodiment and all changes and modifications that fall within the scope of the present invention.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the invention. Thus, provided that these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims (6)

1. The traditional Chinese medicine compound composition for treating liver injury is characterized by comprising the following raw materials in parts by weight: 20-30 parts of purslane, 15-30 parts of cortex albiziae, 15-20 parts of raw malt, 5-10 parts of honey-fried licorice root, 10-20 parts of perilla stem and 15-20 parts of poria cocos.

2. The traditional Chinese medicine compound composition for treating liver injury according to claim 1, which is prepared from the following raw materials in parts by weight: 30 parts of purslane, 30 parts of cortex albiziae, 20 parts of raw malt, 10 parts of honey-fried licorice root, 20 parts of perilla stem and 20 parts of poria cocos.

3. The preparation method of the traditional Chinese medicine compound composition for treating liver injury according to claim 1, which is implemented by the following steps:

step 1, weighing 20-30 parts of purslane, 15-30 parts of cortex albiziae, 15-20 parts of raw malt, 5-10 parts of honey-fried licorice root, 10-20 parts of perilla stem and 15-20 parts of poria cocos for later use;

step 2, crushing the purslane weighed in the step 1, sieving the crushed purslane with a 30-mesh sieve to obtain purslane coarse powder, adding petroleum ether with the weight 10 times that of the purslane coarse powder into the purslane coarse powder, performing reflux extraction for 1 hour to perform degreasing, and performing reduced pressure concentration after the reflux is finished to obtain degreased purslane coarse powder;

adding an ethanol solution which is 10 times of the weight of the defatted purslane coarse powder and has a mass concentration of 80% into the defatted purslane coarse powder, soaking for 0.5h, performing reflux extraction for 2 times after soaking, extracting for 1h each time, filtering after extraction, combining the two filtrates, and performing reduced pressure concentration until no ethanol smell exists to obtain a concentrated solution;

adding distilled water into the concentrated solution, and dispersing the concentrated solution into a liquid medicine containing crude drugs of 0.3 g/ml; loading the liquid medicine on AB-8 type macroporous adsorbent resin, eluting with 2 times column volume of distilled water, and removing water eluate; then eluting with ethanol solution with 4 times column volume and mass concentration of 10%, and discarding the first ethanol eluate; eluting with 80% ethanol solution with 6 times column volume, collecting second alcohol eluate, concentrating under reduced pressure, drying, and pulverizing to obtain herba Portulacae extract;

step 3, mixing the cortex albiziae, the raw malt, the honey-fried licorice root, the perilla stem and the poria cocos weighed in the step 1, adding water which is 5 times of the total weight of the cortex albiziae, the raw malt, the honey-fried licorice root, the perilla stem and the poria cocos into the mixture, soaking the mixture for 30min, decocting the mixture for 1h after soaking, and filtering the decoction to obtain first filtrate and filter residue; adding water with weight 3 times of the residue into the residue, decocting for 0.5 hr, filtering to obtain second filtrate, mixing the first filtrate and the second filtrate, mixing, and concentrating to relative density of 1.2-1.4 to obtain mixed extract;

and 4, uniformly mixing the purslane extract obtained in the step 2 and the mixed extract obtained in the step 3 to obtain the traditional Chinese medicine compound composition for treating liver injury.

4. The use of the compound Chinese medicinal composition for treating liver injury according to any one of claims 1-2 in the preparation of a pharmaceutical preparation for treating liver injury.

5. The application of the traditional Chinese medicine compound composition for treating liver injury in preparing a pharmaceutical preparation for treating liver injury according to claim 4, wherein the traditional Chinese medicine compound composition for treating liver injury is prepared into a pharmaceutically acceptable pharmaceutical preparation according to a conventional method.

6. The application of the traditional Chinese medicine compound composition for treating liver injury in preparing a medicinal preparation for treating liver injury according to claim 5, wherein the medicinal dosage form of the medicinal preparation is oral liquid, capsules, granules, pills, powder, microcapsules or dripping pills.