CN115634245A - Veterinary drug composition, decoction thereof, preparation method and application - Google Patents

Abstract

The invention discloses a veterinary drug composition, a decoction thereof, a preparation method and an application thereof, wherein the composition is prepared from the following raw materials, by weight, 20-40 parts of loosestrife, 15-20 parts of chrysanthemum morifolium and 25-35 parts of thalictrum ramosissimum. The decoction formed by the veterinary drug composition is directed at a target organ of liver, has repairing and treating effects on various forms of acute liver injury, and provides reference data and data support for the administration of liver injury diseases in livestock and poultry breeding.

Description

A kind of veterinary medicine composition and its decoction, preparation method and application

technical field

The invention relates to the field of veterinary medicine, in particular to a veterinary medicine composition and its preparation method and application.

Background technique

Pearl vegetable is the root or whole plant of the Primula family Pearl vegetable, which is bitter, pungent, flat in nature, and enters the liver and spleen meridians. It has the effects of clearing heat and dampness, promoting blood circulation and dispelling blood stasis, detoxifying and eliminating carbuncle, mainly treating edema, hot stranguria, jaundice, dysentery, rheumatism and heat arthralgia, leukorrhea, amenorrhea, bruises, fractures, traumatic bleeding, mastitis, boils, snakes bite.

Chrysanthemum is a perennial herb. According to medical analysis, Chrysanthemum contains ingredients such as Chrysanthemum Chrysanthemum, Chrysanthemum, amino acids and trace vitamins. It is slightly cold in nature and tastes sweet and bitter. It has the functions of dispelling wind and heat, clearing liver and improving eyesight, detoxifying and anti-inflammatory, and can treat hypertension, migraine, acute conjunctivitis and other diseases.

Tangsongcao is the root and rhizome of Tangsongcao in the family Ranunculaceae. It is often used as a Tibetan medicine, and it enters the stomach, large intestine, and liver. Diarrhea, jaundice, leucorrhea, wind-fire toothache, conjunctival congestion and swelling, sores and swelling.

The composition formed by the above three medicines and its therapeutic effects are not disclosed in the prior art.

Contents of the invention

The object of the present invention is to provide a veterinary drug composition and its preparation method and application.

The present invention provides following technical scheme:

A veterinary drug composition, which comprises the following components in parts by mass: 20-40 parts of pearl vegetable, 15-20 parts of chrysanthemum, and 25-35 parts of pine grass.

A decoction of the veterinary composition is also provided.

Further, the decoction of the veterinary drug composition contains the following active ingredients: rutin, luteolin, isochlorogenic acid A and berberine hydrochloride.

The present invention also provides a preparation method of the decoction of the veterinary drug composition, comprising:

S1: adding sterilized deionized water to the components for soaking, and then decocting one or more times to obtain a decoction;

S2: Concentrating the decoction until the concentration of solids is 1-5 g/ml to obtain the decoction.

Further, the solid-to-liquid ratio of the components to the sterilized deionized water is 1:10-1:20.

Further, the decocting temperature is 55-75°C, and the decocting time is 30-50 minutes each time;

The present invention also provides the application of the veterinary drug composition or its decoction in the preparation of veterinary medicine for preventing and treating acute liver injury.

The present invention also provides the application of the veterinary drug composition or its decoction in the preparation of veterinary medicine for preventing and treating chemical acute liver injury caused by carbon tetrachloride.

The present invention also provides the application of the veterinary drug composition or its decoction in the preparation of veterinary drugs for preventing and treating drug-induced acute liver injury caused by acetaminophen.

The invention also provides the application of the veterinary drug composition or its decoction in the preparation of veterinary drugs for preventing and treating alcohol-induced acute liver injury caused by alcohol.

The present invention has the following beneficial effects:

The target organ of the decoction formed by the composition of the present invention is the liver, which has repairing and therapeutic effects on various forms of acute liver injury. The liver damage caused by overdose has a good protective effect, can effectively reduce the serum levels of alanine aminotransferase and aspartate aminotransferase, and can be further used as a drug for preventing and treating liver damage caused by excessive use of drugs in livestock and poultry breeding.

The composition of the invention has good anti-oxidation, antibacterial and anti-inflammation properties, and is non-toxic and harmless to livestock and poultry and the environment.

Description of drawings

FIG. 1 is an HPLC chart of the mixed standard provided in Example 1.

FIG. 2 is an HPLC chart of the decoction of the composition provided in Example 1. FIG.

Fig. 3 is the section of organs of the toxicity test provided in Example 1 (HE staining, ×400).

FIG. 4 is a diagram of the changes in appearance of acute liver injury caused by CCl ₄ provided in Example 1. FIG.

Fig. 5 is the histopathological changes of liver with acute liver injury caused by CCl ₄ provided in Example 1 (HE staining, ×400).

FIG. 6 shows the detection results of serum biochemical indicators in mice with CCl ₄ -induced acute liver injury provided in Example 1.

FIG. 7 shows the liver antioxidant capacity of mice with acute liver injury caused by CCl ₄ provided in Example 1.

FIG. 8 shows the detection results of liver inflammatory factors in mice with acute liver injury caused by CCl ₄ provided in Example 1.

FIG. 9 shows the detection results of miR-122 expression in the liver of mice in the CCl ₄ -induced acute liver injury model group provided in Example 1.

FIG. 10 shows the visual changes of the liver in acute liver injury caused by APAP provided in Example 1.

Fig. 11 shows the histopathological changes of acute liver injury induced by APAP provided in Example 1 (HE staining, ×400).

FIG. 12 shows the detection of serum biochemical indicators in mice with APAP-induced acute liver injury provided in Example 1. FIG.

FIG. 13 is the detection results of liver antioxidant capacity indicators in mice with APAP-induced acute liver injury provided in Example 1. FIG.

FIG. 14 is the detection results of liver inflammatory factors in mice with acute liver injury caused by APAP provided in Example 1. FIG.

FIG. 15 shows the expression level of miR-122 in the liver of mice with APAP-induced acute liver injury provided in Example 1.

FIG. 16 shows changes in liver autopsy for acute liver injury caused by Alcohol provided in Example 1.

Fig. 17 shows the histopathological changes of liver with acute liver injury caused by Alcohol provided in Example 1 (HE staining, ×400).

FIG. 18 shows the detection results of serum biochemical indicators in mice with Alcohol-induced acute liver injury provided in Example 1. FIG.

FIG. 19 shows the anti-oxidative capacity of the liver in acute liver injury caused by Alcohol provided in Example 1.

FIG. 20 shows the detection results of liver inflammatory factors in mice with Alcohol-induced acute liver injury provided in Example 1.

FIG. 21 shows the detection results of miR-122 expression in the liver of mice with Alcohol-induced acute liver injury provided in Example 1.

Fig. 22 is the effect of the composition decoction provided by the present invention in Example 2 on alanine aminotransferase and aspartate aminotransferase of young yaks with liver injury caused by praziquantel.

Detailed ways

The present invention will be described in detail below in conjunction with the embodiments and drawings, but it should be understood that the embodiments and drawings are only used for exemplary description of the present invention, and do not constitute any limitation on the protection scope of the present invention. All reasonable transformations and combinations within the scope of the gist of the present invention fall within the protection scope of the present invention.

The test methods used in the following examples include:

1. Methods for evaluating the antioxidant effect in vitro:

(1) Total antioxidant capacity of Fe

Take 10 μL of 1 g/mL composition decoction, add 20 μL of pH 6.6 phosphate buffer solution and 20 μL of 1% potassium ferricyanide solution, mix and place at 50°C for 20 minutes, add 250 μL of 10% trichloroacetic acid solution and mix, and take the mixture 25 μL, add 25 μL of distilled water and 25 μL of 0.1% ferric chloride solution, mix well, let it stand for 10 minutes, measure the optical density OD at 700 nm, and use the solvent instead of the composition decoction as a blank control.

The total antioxidant capacity of Fe is represented by ΔOD, ΔOD=OD sample-OD blank.

(2) Hydroxyl radical scavenging rate

Add 0.5mL of 2mmol/L ferrous sulfate solution and 0.5mL of 6mmol/L hydrogen peroxide to the test tube and shake well, then add 1.5mL of 6mmol/L salicylic acid, take it out in 37℃ water bath for 15min, measure its optical density OD, add 0.1 mL of composition decoction to the blank group and shake well, continue heating in the water bath for 15 minutes, take out 200 μL of the OD sample to measure its optical density, and obtain the hydroxyl radical scavenging rate according to the following formula:

Clearance rate (%)=(OD blank-OD sample)/OD blank×100%

(3) DPPH clearance rate

Prepare DPPH as a 0.5 mg/mL solution, store in the dark at 4°C, and finish the test within 3.5 hours.

Add 100 μL/well of 1 g/mL composition decoction into the microwells of the microplate, and then add 100 μL of DPPH solution therein, and set 3 replicate wells. In the blank control group and the background group, 100 μL of pure water and absolute ethanol were used to replace the DPPH solution, and three replicate wells were also set. Stand at 37°C for 30 minutes in the dark, and measure the OD value at 517nm with a microplate reader.

The DPPH clearance rate was obtained by the following calculation formula:

DPPH clearance rate (%) = [OD blank - (OD sample - OD background)] / OD blank × 100%

2. Anti-inflammatory test method:

The in vitro anti-inflammatory activity of composition decoction is characterized by the inhibition rate of 5-lipoxygenase activity measured by spectrophotometry, wherein:

Preparation of the substrate: Disperse Tween-20 in 0.02mol/L, pH 9.0 borate buffer solution, add an appropriate amount of linoleic acid during the shaking process, and then dropwise add 1 mol/L sodium hydroxide solution to make It became clear and the pH remained at 9.0.

Take 1 μL of the composition decoction with a concentration of 1g/mL and add it to a 1.5mL EP tube containing 20 μL of lipoxygenase solution, heat in a water bath at 30°C for 30 minutes, add 150mL of the substrate, put it in a water bath at 30°C for 3 minutes, add 500 μL of absolute ethanol was added to terminate the reaction and mixed with 500 μL of distilled water to be tested, which was recorded as OD1; in the positive control group, 1 μL of distilled water was used instead of the composition decoction, which was recorded as OD2; in the negative control group, 20 μL of distilled water was used instead of lipoxygenase, which was recorded as OD3; In the blank control group, 1 liter of distilled water was used to replace the composition decoction, and 500 μL of absolute ethanol was added to terminate the reaction before adding the substrate, which was recorded as OD4; the test was repeated 3 times, and 200 μL of the composition decoction was taken to detect the absorbance of each reaction solution at 234 nm. OD value,

The anti-inflammatory rate was obtained by the following formula:

Anti-inflammatory rate (%)=[1-(OD1-OD3)/(OD2-OD4)]×100%

3. Bacteriostasis test method:

Use the Oxford cup antibacterial method: after diluting Escherichia coli ATCC25922 and Staphylococcus aureus ATCC43300 bacteria solution in a certain proportion, spread the spreader evenly on the MH agar medium, and then place the Oxford cup on the MH agar medium , take 200 μL of the composition decoction and add it into an Oxford cup, and measure the size of the inhibition zone after constant temperature incubation at 37°C.

Four, composition decoction ingredient analysis:

Taking rutin, luteolin, isochlorogenic acid A and berberine hydrochloride as reference components, an HPLC-based qualitative and quantitative analysis method for the main components of the composition decoction and its content was established to analyze the pharmacological effects of the composition decoction material basis. The specific process includes:

Accurately weigh an appropriate amount of rutin, luteolin, isochlorogenic acid A, and berberine hydrochloride standard substances, prepare mother liquors with chromatographic grade methanol respectively, and dilute to a suitable concentration before use and filter through a 0.22 μm microporous membrane. The chromatographic conditions are as follows: AgilentSB-C18 chromatographic column (4.6mm×250mm, 1.5μm), mobile phase A is acetonitrile, B is 0.2% phosphoric acid aqueous solution, gradient elution: 0～11min, A phase 16%～18%; 11- 25min, A phase 18%～25%, 25～30min; A phase 25%～30%; 30～40min, A phase 30%～35%; 40～50min, A phase 35%～40%; 50～55min, Phase A 40%-16%; flow rate 1mL/min, detection wavelength 348nm, injection volume 10μL, column temperature 30°C.

5. Acute toxicity and cumulative toxicity test:

42 18-22g female BALB/c mice were randomly divided into 7 groups, respectively blank control group (Control), A, B, C, D, E, F groups, blank control group was given normal saline, A, Groups B, C, D, E, and F were respectively based on crude drug concentrations of 30g/kg·bw, 20g/kg·bw, 13.3g/kg·bw, 8.9g/kg·bw, 5.9g/kg·bw, 3.9g/kg/ kg·bw dose orally, observed for 24 hours. If there is no death or other abnormal phenomena, continue to carry out the 21-day cumulative toxicity test, and conduct an autopsy on the 22nd day to observe the general status, organ index, organ pathology and histopathology of the heart, liver, spleen, lung, kidney and other solid organs Variety.

6. Evaluation test for the protective effect on acute liver injury:

From 113 18-22g male BALB/c mice, 8 were randomly selected as the blank control group, and the remaining 105 were divided into CCl4 (carbon tetrachloride) group, APAP (acetaminophen) group, and Alcohol (alcohol) group , 35 rats in each group, and each group was randomly divided into 5 groups, namely model group (Model), silybin group (Silybin), high-dose group (H), middle-dose group (M), low-dose group group (L). The blank control group was given continuous intragastric administration of normal saline for 14 days; the model group was not given intragastric administration of drugs; the silibinin group was given intragastric administration of 50 mg/kg for 14 consecutive days; Concentrations of 10g/kg·bw, 5g/kg·bw, 2.5g/kg·bw were administered continuously for 14 days. On day 15, except for the blank control group, the liver injury model was replicated in other groups according to the following model replication method.

Replication of the chemical acute liver injury model (carbon tetrachloride, CCl4): intraperitoneally inject 1% carbon tetrachloride solution at 10 g/kg bw, kill all test mice on the 16th day and perform autopsy;

Drug-induced acute liver injury model replication (acetaminophen, APAP): 200mg/kg bw intraperitoneal injection of acetaminophen solution, all experimental mice were killed on the 16th day and autopsyed;

Alcohol-induced acute liver injury model replication (alcohol, Alcohol): 10ml/kg·d continuous gavage of white wine with an alcohol content of 53 degrees for 5 days, all experimental mice were killed on the 20th day and autopsy was performed.

Among them, the evaluation indicators of acute liver injury are as follows

(1) Organ index

Rinse the mouse heart, liver, spleen, lung, and kidney with sterilized normal saline, dry them with filter paper, weigh them and calculate the organ index: organ index = organ/body weight × 100%

(2) Histopathological observation of the liver

Take the middle part of the left lobe of the liver, fix it in 4% paraformaldehyde for 48 hours, then carry out paraffin embedding, sectioning, dewaxing, hematoxylin-eosin (HE) staining, and finally observe its pathological morphology under the microscope Variety.

(3) Detection of serum biochemical indicators

Serum TG and T-CHO levels were determined according to the instructions of commercial serum biochemical kits.

(4) Evaluation of liver antioxidant capacity and analysis of related gene expression levels

After the liver homogenate was prepared, T-AOC, GSH-Px, SOD, and MDA were tested according to the instructions of the commercial kit.

Weighed 50 mg of liver tissue homogenate, added 1 mL of Trizol lysate to extract RNA, detected the RNA concentration with an ultra-micro nucleic acid protein analyzer, reverse-transcribed RNA into cDNA on a gradient PCR instrument, and then measured the inflammatory factor IL- The relative expression of 1β, IL-6, TNF-α and liver microRNA miR-122. GAPDH and U6 were selected as internal reference genes, and the primer sequences are shown in Table 1.

Table 1 Primer sequences for qPCR detection

7. Tests on the effect of praziquantel on liver damage in young yaks:

21 healthy young yaks (blank control group, praziquantel group and composition decoction group) were selected to conduct a clinical evaluation of the protective effect of the composition decoction on young yak liver damage caused by excessive use of praziquantel. Except for the blank group, the other 14 young yaks were orally administered praziquantel at a dose of 50 mg/kg·bw for 14 consecutive days, while the composition decoction group was orally administered with a dose of 10 g/kg·bw. On the 15th day, blood was collected from the jugular vein to determine the contents of alanine aminotransferase and aspartate aminotransferase in serum. During the experiment, the appearance and physiological responses of young yaks in each group were closely observed.

The statistics and analysis of the test results are as follows:

表示。组间统计分析采用LSD检验，P<0.05为差异有统计学意义，记为*；P<0.01，记为**；P<0.001，记为***。SPSS software was used for statistical processing of data, and the test results were

express. Statistical analysis among groups was performed by LSD test. P<0.05 indicated a statistically significant difference, marked as *; P<0.01, marked as **; P<0.001, marked as ***.

Example 1

Prepare groups of decoctions as follows:

S1: Mix the components of the composition with the water, which is sterilized deionized water, at a solid-to-liquid ratio of 1:10, soak for 30 minutes, and decoct twice, each time for 45 minutes;

S2: Collect the filtrate after filtration, and concentrate the filtrate to 1 g/mL (part of the concentrated solution is sterilized by filtering through a 0.22 μm syringe disposable filter) to obtain a decoction.

Wherein, the compositions in different groups of decoctions meet the following ingredients and proportioning (table 2):

Table 2 Orthogonal design level factor table

The in vitro antioxidant, anti-inflammatory and bacteriostatic properties of different groups of pharmaceutical compositions were tested. Among them, in the antioxidant test, the total antioxidant capacity of Fe, hydroxyl radical scavenging rate (%) and DPPH scavenging rate (%) of pearl vegetable were 3.67±0.02, 61.07±7.40 and 90.98±1.45, respectively, and the Fe of Hangju The total antioxidant power, hydroxyl radical scavenging rate (%) and DPPH scavenging rate (%) were 1.17±0.02, 66.84±2.46 and 70.77±20.84, respectively. (%) and DPPH clearance rate (%) were 2.41 ± 0.32, 48.77 ± 0.76 and 69.85 ± 5.29 respectively; in the anti-inflammatory test, the anti-inflammatory rate (%) of pearl vegetable, hangchrysanthemum, and syringa spp. 0.46, 78.54±8.94 and 64.71±5.84; in the antibacterial test, the antibacterial zone sizes of pearls against ATCC 25922 and ATCC 43300 were 14.1±0.2mm and 13.7±0.4mm respectively, and the antibacterial effects of Hangju on ATCC 25922 and ATCC 43300 The sizes of the circles were 12.3±0.3mm and 11.6±0.5 mm, respectively, and the antibacterial zone sizes of P. spp. against ATCC 25922 and ATCC 43300 were 13.5±1.1 and 12.3±0.3mm, respectively.

It can be seen from Table 3 that the influence of the three factors in the antioxidant test is ranked as B>A>C in descending order, and it is deduced that the combination of anti-oxidation-dominated decoction is A1B1C2, which is recorded as ZJD1.

Table 3 The range analysis and comprehensive score dominated by antioxidant

A B C Overall rating 1 1 1 1 twenty four 2 1 2 2 19 3 1 3 3 17 4 2 1 2 18 5 2 2 1 14 6 2 3 3 twenty three 7 3 1 2 twenty two 8 3 2 1 19 9 3 3 3 17 K1 60 64 57 K2 55 52 59 K3 58 57 57 k1 20 21.33 19 k2 18.33 17.33 19.67 k3 19.33 19 19 R 1.67 4 0.67

It can be seen from Table 4 that the influence of the three factors in the anti-inflammatory test is ranked from large to small as A>B>C, and it can be deduced that the composition decoction combination dominated by inflammation is A ₁ B ₁ C ₂ , which is recorded as ZJD ₁ (It is the same as the drug combination confirmed by anti-oxidation-based comparison, which are all recorded as ZJD ₁ ).

Table 4 The range analysis and comprehensive score dominated by anti-inflammation

It can be seen from Table 5 that the influence of the three factors in the antibacterial test is ranked from large to small as A>C>B, and it is deduced that the antibacterial-dominated composition decoction combination is A ₃ B ₃ C ₁ , which is recorded as ZJD ₂ .

Table 5 The antibacterial effect is the leading range analysis and comprehensive score

A B C Bacteriostatic Composite Score 1 1 1 1 9 2 1 2 2 6 3 1 3 3 6 4 2 1 2 4 5 2 2 1 4 6 2 3 3 4 7 3 1 2 5 8 3 2 1 8 9 3 3 3 9 K1 21.0 18.0 21.0 K2 14.0 18.0 15.0 K3 22.0 19.0 19.0 k1 7.0 6.0 7.0 k2 4.7 6.0 5.0 k3 7.3 6.3 6.3 R 2.7 0.3 2.0

Finally, with anti-oxidation, anti-inflammatory, and antibacterial comprehensive activities as the main factors, the composition decoction combinations with the highest scores for anti-oxidation, anti-inflammatory, and antibacterial comprehensive activities were compared, as shown in Table 6, and finally ZJD3 was determined to be the most effective composition decoction. Good composition.

Table 6 Range analysis and comprehensive score dominated by antioxidant, anti-inflammatory and antibacterial comprehensive activities

The composition decoction prescription ZJD3 (A1B1C3) was repeated three times to verify the repeatability, as shown in Table 7.

Table 7 Composition decoction in vitro activity repeatability evaluation test

Further, based on HPLC (High Performance Liquid Chromatography), the main components of the composition decoction and their content are analyzed, wherein the mixed standard linear regression equation is as shown in Table 8, and the liquid chromatogram of the standard and composition decoction They are shown in Figure 1 and Figure 2 respectively.

According to the linear regression analysis, every 1 g of the composition decoction contains 0.27 mg of rutin, 2.72 mg of luteolin, 0.26 mg of isochlorogenic acid A, and 0.28 mg of berberine hydrochloride.

Table 8 standard curve

Further, the meridian distribution of the composition decoction was analyzed by the pharmacological experiment method of traditional Chinese medicine

The meridian analysis results showed that the rate of the composition decoction entering the liver meridian reached 100%, indicating that the composition decoction had preventive and therapeutic effects on liver diseases, as shown in Table 9.

Table 9 Composition Decoction Meridian Analysis

Further, the acute toxicity and cumulative toxicity of the composition decoction were tested and evaluated. Among them, the heart, liver, spleen, lung, kidney and other organ indexes of the mice administered with the composition decoction group were not significantly different from those of the blank control group. difference, the obtained results are shown in Table 10 and Figure 3, showing that the composition decoction is safe and nontoxic.

Table 10 Toxicity test organ index

Further, the effect of the composition decoction on acute liver injury caused by CCl ₄ is to study the preliminary evaluation of the protective effect of the composition decoction on acute liver injury.

(1) Organ index:

Compared with the blank control group, after intraperitoneal injection of CCl ₄ for model replication, the liver index decreased, and improved after treatment with the composition decoction. The liver index of the silibinin group was close to that of the blank control group, and there was a significant difference from the model group (See Table 11).

Table 11 CCl ₄ induced acute liver injury mouse organ index

As can be seen in Table 11, a: there is a significant difference compared with the blank control group; b: there is a significant difference compared with the model group.

(2) Histopathological changes of the liver

Autopsy of the CCl ₄ acute liver injury model group showed red hemorrhagic spots and darker color in the liver; different degrees of white necrosis appeared in the silibinin group, middle dose group and low dose group, and the surface of the high dose group was smooth without obvious Changes in appearance, see Figure 4.

In the blank control group, the hepatic cord structure was clear and the hepatocytes were arranged neatly. In the model group, the hepatic cord was disordered, the hepatic sinusoid was narrowed, and a large area of liver fibrosis appeared around it. Compared with the model group, the swelling, deformation and necrosis of liver cells in each dose group were significantly reduced, as shown in Figure 5.

(3) Detection results of serum biochemical markers in mice with acute liver injury caused by CCl4

The detection results of serum biochemical indicators showed that after 14 days of continuous gavage of the composition decoction, the AST and ALT of the high-dose composition decoction group tended to be consistent with those of the blank control group. Each dose group of the composition decoction has different degrees of inhibitory effects on the expression of TG, and the silibinin group and the high dose group of the composition decoction have a maintenance effect on T-CHO.

(4) Antioxidant effect

Compared with the blank control group, the SOD, GSH-Px, and T-AOC in the liver tissue of the mice in the model group were significantly reduced, while the content of MDA was significantly increased. Compared with the model group, SOD, CAT, GSH-Px and MDA in the liver tissue of the silybin group and the composition decoction group were all improved to varying degrees, as shown in Figure 7.

(5) Detection results of liver inflammatory factors in mice with acute liver injury caused by CCl4

The qPCR test results showed that, compared with the model group, each dose group of the composition decoction had a certain inhibitory effect on the expression of IL-1β, IL-6, and TNF-α, and it was statistically significant, as shown in Figure 8.

(6) Detection results of liver miR-122 in mice with acute liver injury caused by CCl4

Compared with the blank control group, the expression of miR-122 in the model group was significantly increased. It can be concluded from the results that the abnormal expression of miR-122 can be improved in the H and M dose groups after oral administration of the composition decoction, as shown in Figure 9.

Further, the composition decoction is tested for the impact on BALB/c mice with acute liver injury caused by APAP:

(1) Organ index

Compared with the model group, after intragastric administration of the composition decoction, the liver indexes of the H, M, and L dose groups showed significant differences, indicating that different dose groups of the composition decoction had protective effects on acute liver injury caused by APAP. (Table 12).

Table 12 APAP-induced acute liver injury mouse organ index

As can be seen in Table 12, a: there is a significant difference compared with the blank control group; b: there is a significant difference compared with the model group

(2) Liver pathological changes in eye appearance

The livers of the mice in the silibinin group were congested to varying degrees. No obvious pathological changes appeared on the surface of the livers of the mice at the doses of composition decoctions H and M, which were normally dark red and soft in texture, as shown in FIG. 10 .

HE staining showed that the hepatic necrosis area was significantly reduced after intragastric administration in each dose group of the composition decoction, and the degeneration and necrosis of liver tissue was alleviated, indicating that the intervention of the composition decoction can improve the acute liver injury caused by APAP, as shown in Figure 11.

(3) Detection of serum biochemical markers in mice with acute liver injury induced by APAP

Each dose group of the composition decoction reduced the influence of AST, ALT, TG, T-CHO APAP model replication to varying degrees, and basically recovered to the level of the blank control group, as shown in Figure 12.

(4) Liver antioxidant capacity of rats with acute liver injury induced by APAP

Compared with the model group, after intragastric administration of the composition decoction, the abnormalities of MDA, SOD, GSH-Px, and T-AOC caused by APAP model replication were alleviated to varying degrees, as shown in Figure 13.

(5) Detection of liver inflammatory factors in mice with acute liver injury caused by APAP

Compared with the model group, the different dosage groups of the composition decoction had inhibitory effects on the expression of IL-1β, IL-6, and TNF-α in APAP-induced acute liver injury, with statistical significance, as shown in FIG. 14 .

(6) The effect of APAP-induced acute liver injury on the expression of miR-122 in the liver of mice

Compared with the blank control group, the expression of miR-122 in the model group was significantly increased, and the expression of miR-122 in the composition decoction H and M dose groups tended to the blank control group, as shown in Figure 15.

Further, the impact test of the composition decoction on Alcohol-induced acute liver injury BALB/c mice:

(1) Organ index

The liver index of the silibinin group, the composition decoction H, and the middle dose group showed significant differences from the model group, as shown in Table 13.

Table 13 Alcohol-induced acute liver injury mouse organ index

As can be seen in Table 13, a: there is a significant difference compared with the blank control group; b: there is a significant difference compared with the model group.

(2) Observation of liver pathological changes

Compared with the blank control group, the high-dose composition decoction group had no obvious appearance changes in the liver, the middle-dose group had some inconspicuous white necrosis, and the low-dose group had congestion around the liver, as shown in Figure 16.

After HE staining in the blank control group, the liver cells were in a healthy state, and there was no degeneration, necrosis and other pathological changes in the liver. After intragastric administration of the composition decoction, the degree of liver cell injury in each dose group was alleviated, and the relief was more obvious in group H, as shown in Figure 17.

(3) Detection results of serum biochemical markers in mice with acute liver injury caused by Alcohol

Compared with the model group, both the silibinin group and the high-dose composition decoction group had significant and statistically significant improvement effects on abnormal serum indicators AST, ALT, TG, and T-CHO, as shown in Figure 18.

(4) Antioxidative capacity of liver in mice with acute liver injury induced by Alcohol

There were significant differences between Alcohol-induced acute liver injury mice and the blank control group. Each dose group of the composition decoction reversed the effects of MDA, SOD, GSH-Px, and T-AOC after the APAP model was replicated to varying degrees. Abnormal, see Figure 19.

(5) Detection results of liver inflammatory factors in mice with Alcohol acute liver injury

Compared with the model group, different doses of the composition decoction had a certain inhibitory effect on the expression of IL-1β, IL-6, and TNF-α in the mice with acute liver injury caused by Alcohol, as shown in Figure 20 .

(6) Detection results of liver miR-122 in mice with Alcohol acute liver injury

Compared with the blank control group, the expression of miR-122 in the model group was significantly increased, and the abnormal expression of miR-122 was significantly improved in the high-dose group after oral administration of the composition decoction, as shown in Figure 21.

Example 2

The test of the efficacy of the ZJD3 (A1B1C3) composition decoction on the liver damage of young yaks caused by praziquantel:

After continuous administration of the ZJD3 (A1B1C3) composition decoction in Example 1, that is, the composition decoction provided by the present invention for 14 days, the contents of serum alanine aminotransferase and aspartate aminotransferase in young yaks were obtained, as shown in Figure 22 (* in the figure represents that there is a significant difference compared with the blank control group; # represents that there is a significant difference compared with the praziquantel group), wherein figure (a) is the content situation of alanine aminotransferase, and figure (b) is the content of aspartate aminotransferase It can be seen that the contents of serum alanine aminotransferase and aspartate aminotransferase in young yaks after administration were significantly lower than those in the praziquantel group. Compared with the blank control group, there was no significant difference in the content of alanine aminotransferase. In addition, during the experiment, two young yaks in the praziquantel group were found to have different degrees of muscle tremors, while neither the blank group nor the composition decoction group had this phenomenon. The above results indicated that the composition decoction could alleviate adverse reactions such as elevated serum alanine aminotransferase and aspartate aminotransferase and muscle tremors in young yaks caused by praziquantel, and had a protective effect on young yak liver damage caused by praziquantel.

The above examples are only preferred implementations of the present invention, and the scope of protection of the present invention is not limited to the above examples. All technical solutions under the idea of the present invention belong to the protection scope of the present invention. It should be pointed out that for those skilled in the art, improvements and modifications without departing from the principle of the present invention should also be regarded as the protection scope of the present invention.

Claims (10)

1. A veterinary drug composition, characterized in that it comprises the following components in parts by mass: 20-40 parts of pearl vegetable, 15-20 parts of chrysanthemum, and 25-35 parts of pine grass. 2. the decoction of the described veterinary medicine composition of claim 1. 3. The decoction of the veterinary drug composition according to claim 2, characterized in that it contains the following active ingredients: rutin, luteolin, isochlorogenic acid A and berberine hydrochloride. 4. according to the preparation method of the decoction of described veterinary drug composition of claim 2 or 3, it is characterized in that, it comprises: S1: adding sterilized deionized water to the components for soaking, and then decocting one or more times to obtain a decoction; S2: Concentrating the decoction until the concentration of solids is 1-5 g/ml to obtain the decoction. 5. The preparation method according to claim 4, characterized in that, the solid-to-liquid ratio of the components to the sterilized deionized water is 1:10˜1:20. 6 . The preparation method according to claim 4 , characterized in that, the decocting temperature is 55-75° C., and the decocting time is 30-50 minutes each time. 7. The application of the decoction of the veterinary drug composition according to claim 1 or the veterinary drug composition according to claim 2 or 3 in the preparation of veterinary medicines for preventing and treating acute liver injury. 8. according to claim 1 described veterinary drug composition or the application of the decoction of claim 2 or 3 described veterinary drug composition in the medicine of the chemical acute liver injury that prevention and treatment is caused by carbon tetrachloride for preparation. 9. Use of the veterinary drug composition according to claim 1 or the decoction of the veterinary drug composition according to claim 2 or 3 in the preparation of veterinary medicines for preventing and treating drug-induced acute liver injury caused by acetaminophen. 10. Use of the decoction of the veterinary drug composition according to claim 1 or the veterinary drug composition according to claim 2 or 3 in the preparation of veterinary medicines for preventing and treating alcohol-induced acute liver injury caused by alcohol.

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