阅读说明：本技术 一种用于治疗犊牛胃肠道疾病的藏药复方及其制备方法和应用 (Tibetan medicine compound for treating calf gastrointestinal diseases and preparation method and application thereof ) 是由 张凯 张康 仇正英 王磊 张景艳 王贵波 郭志廷 王学智 李建喜 于 2021-01-13 设计创作，主要内容包括：本发明涉及一种用于治疗犊牛腹泻的藏药复方及其制备方法,涉及医药技术领域。所述的藏药复方由下述重量配比的原料制成：三颗针1～3份、沙棘1～4份、诃子5份、甘青青兰2份、矮紫堇2份。本发明的藏药复方具有抑菌止泻镇痛功效,同时还具备降低肠推进率的作用,抑制肠上皮细胞脱落,降低粘膜水肿、减少炎症细胞浸润,促进腹泻快速愈合,不易产生细菌耐药性,不影响动物生产性能。(The invention relates to a Tibetan medicine compound for treating calf diarrhea and a preparation method thereof, and relates to the technical field of medicines. The Tibetan medicine compound is prepared from the following raw materials in parts by weight: 1-3 parts of radix berberidis, 1-4 parts of sea buckthorn, 5 parts of myrobalan, 2 parts of dracocephalum tanguticum and 2 parts of corydalis tuber. The Tibetan medicine compound disclosed by the invention has the effects of inhibiting bacteria, relieving diarrhea and easing pain, and also has the effect of reducing the intestinal propulsion rate, inhibiting the shedding of intestinal epithelial cells, reducing mucosal edema, reducing inflammatory cell infiltration, promoting the quick healing of diarrhea, and is not easy to generate bacterial resistance and not influence the production performance of animals.)

1. A Tibetan medicine compound for treating calf gastrointestinal diseases is characterized by being prepared from the following raw materials in parts by weight: 1-3 parts of radix berberidis, 1-4 parts of sea buckthorn, 5 parts of myrobalan, 2 parts of dracocephalum tanguticum and 2 parts of corydalis tuber.

2. The Tibetan medicine compound of claim 1, which is prepared from the following raw materials in parts by weight: 2-3 parts of radix berberidis, 1-4 parts of sea buckthorn, 5 parts of myrobalan, 2 parts of dracocephalum tanguticum and 2 parts of corydalis tuber.

3. The Tibetan medicine compound of claim 2, which is prepared from the following raw materials in parts by weight: 2 parts of root of Chinese barberry, 4 parts of sea buckthorn, 5 parts of myrobalan, 2 parts of dracocephalum tanguticum and 2 parts of corydalis tuber.

4. The Tibetan medicine compound of claim 2, which is prepared from the following raw materials in parts by weight: 2 parts of radix berberidis, 3 parts of sea buckthorn, 5 parts of myrobalan, 2 parts of dracocephalum tanguticum and 2 parts of corydalis tuber.

5. The Tibetan medicine compound of any one of claims 1 to 4, which is prepared into decoction, tablets, granules, pills or oral liquid by adding pharmaceutically acceptable auxiliary materials.

6. The Tibetan medicine compound of claim 5, which is prepared into oral liquid by adding pharmaceutically acceptable auxiliary materials.

7. A preparation method of the Tibetan medicine compound oral liquid as claimed in any one of claims 1 to 4, which is characterized by comprising the following steps:

(1) weighing radix Berberidis, fructus Hippophae, fructus Chebulae, herba Dracocephali and herba corydalis edulis according to a certain proportion, and cleaning with cold water to remove floating dust;

(2) soaking the medicinal materials obtained in the step (1) in cold water;

(3) adding the soaked medicinal materials in the step (2) into a closed decoction machine, adding water with the volume of 5-10 times of that of the medicinal materials, and decocting the medicinal materials for 2 hours at 115 ℃;

(4) collecting the liquid medicine decocted in the step (3), and filtering four layers of gauze to obtain filtrate;

(5) adding 5-10 times of water, repeating the step (3), decocting with water at 115 deg.C for 2 hr;

(6) collecting the liquid medicine decocted in the step (5), and filtering the liquid medicine with four layers of gauze to obtain filtrate;

(7) mixing the two filtrates obtained in the steps (4) and (6), adding into a rotary evaporator, concentrating the medicinal liquid at 0.05Mpa and 60 deg.C until 1 ml of medicinal liquid is equal to 1g of the original medicinal material;

(8) and (5) adding 0.2% of potassium sorbate into the concentrated liquid medicine in the step (7), canning and sealing.

8. Use of the Tibetan medicine compound as claimed in any one of claims 1 to 4 in the preparation of a medicament for treating gastrointestinal diseases of calves.

9. Use of the Tibetan medicine compound of any one of claims 1-4 in the preparation of a medicament for treating colitis in calves.

10. Use of the Tibetan medicine compound of any one of claims 1-4 in the preparation of a medicament for treating calf diarrhea.

Technical Field

The invention relates to the technical field of medicinal preparations, in particular to a Tibetan medicine compound for treating gastrointestinal diseases of calves and a preparation method and application thereof.

Background

The gastrointestinal tract diseases comprise diseases of esophagus, stomach, small intestine, colon, rectum and the like, and common main symptoms comprise rhythmicity, periodic upper abdominal pain, diarrhea, hunger abdominal pain, acid regurgitation, fever, black stool and bloody stool, gastrointestinal bleeding, intestinal obstruction and the like. Gastrointestinal disorders are one of the most common diseases in humans, the most common of which include swallowing disorders, gastric ulcers, peptic ulcers, gastroparesis, delayed gastric emptying, Irritable Bowel Syndrome (IBS), and Inflammatory Bowel Disease (IBD). Diarrhea is a symptom of intestinal diseases, is mostly caused by disease factors, and is common in intestinal diseases, other common diseases include acute food poisoning, systemic infection and the like, and bacterial infection is one of common causes. Calf diarrhea seriously affects the healthy development of the cattle breeding industry. Calf diarrhea is caused by infection of pathogenic microorganisms such as bacteria, viruses and parasites, or various factors such as nutritional factors, environment and management. In recent years, along with the scale expansion of the dairy cow breeding industry, the incidence rate of calf diarrhea is in an increasing trend, and the calf diarrhea seriously affects the early growth and development of calves and the exertion of later-stage production performance. The calf diarrhea not only directly affects the growth and development of calves, but also affects the renewal of cattle herds and the benign development of the dairy cow breeding industry due to high mortality. Meanwhile, some protozoa causing calf diarrhea are important zoonosis microorganisms, and can seriously threaten the public health and safety of regions. Therefore, the calf diarrhea can be effectively controlled.

Calf diarrhea generally occurs in calves within 1 month. It is characterized clinically as diarrhea and dyspepsia and is a gastrointestinal digestive disease. Calf diarrhea can occur all the year round, especially the calf is more prone to occur around the air temperature, when the calf is in the lactation period, the calf is more prone to attack diseases, the incidence rate of the calf diarrhea accounts for about 80% of the incidence rate of the whole calf, and even if the calf suffering from the calf diarrhea is easy to infect respiratory diseases in the growth process compared with other calves. Therefore, the disease has great influence on the growth and development and survival of calves.

Calf diarrhea has complex causes, is difficult to take medicine according to symptoms, has high death rate, and is called as a killer of a newborn calf. Once the disease is developed, antibiotics such as gentamicin, norfloxacin, oxytetracycline, norfloxacin and the like are mostly used for treating the disease, but the disease is not necessarily caused by bacteria, so that the pertinence is not strong, the problems of bacterial drug resistance, excessive corresponding animal product drug residue and the like are easy to occur, and the problems of serious public health safety and the like are caused. The skilled in the art has made a lot of studies on gastrointestinal diseases of calves, especially calf diarrhea, for example, patent CN201410028836.2 discloses a Tibetan medicine composition for treating yak calf diarrhea, which comprises the following raw material components: 375-420 parts of cichorium endivia, 320-350 parts of phellodendron, 145-160 parts of pomegranate rind and 100 parts of kaempferia galanga. The pharmaceutical composition has the characteristics of obvious clinical curative effect, low side effect, no residue, low cost, convenient use and the like, and has good treatment effect on diarrhea of yak calves. Meanwhile, due to the adoption of the form of the traditional Chinese medicine composition, in the treatment of the disease, the use amount of antibiotics and chemosynthesis medicines in the treatment of diarrhea of yak calves can be effectively reduced, the threat of medicine residues on food safety and public health is eliminated, and the social requirements of providing safe and pollution-free animal-derived food and human health are met. However, the effects of the drugs such as bacteriostasis, analgesia, anti-inflammation and the like are not studied in detail. Aiming at the technical problems, the invention provides a Tibetan medicine compound for treating gastrointestinal tract diseases of calves. The Tibetan medicine compound of the invention can quickly stop diarrhea, reduce the use of antibiotics and reduce the generation of bacterial drug resistance.

Disclosure of Invention

The invention aims to provide a Tibetan medicine compound for treating calf diarrhea and a preparation method thereof through a large number of experimental studies. The specific technical scheme is as follows:

a Tibetan medicine compound for treating calf gastrointestinal diseases is prepared from the following raw materials in parts by weight: 1-3 parts of radix berberidis, 1-4 parts of sea buckthorn, 5 parts of myrobalan, 2 parts of dracocephalum tanguticum and 2 parts of corydalis tuber.

Preferably, the Tibetan medicine compound is prepared from the following raw materials in parts by weight: 2-3 parts of radix berberidis, 1-4 parts of sea buckthorn, 5 parts of myrobalan, 2 parts of dracocephalum tanguticum and 2 parts of corydalis tuber.

Preferably, the Tibetan medicine compound is prepared from the following raw materials in parts by weight: 2 parts of root of Chinese barberry, 4 parts of sea buckthorn, 5 parts of myrobalan, 2 parts of dracocephalum tanguticum and 2 parts of corydalis tuber.

Preferably, the Tibetan medicine compound is prepared from the following raw materials in parts by weight: 2 parts of radix berberidis, 3 parts of sea buckthorn, 5 parts of myrobalan, 2 parts of dracocephalum tanguticum and 2 parts of corydalis tuber.

Preferably, the Tibetan medicine compound is added with pharmaceutically acceptable auxiliary materials to prepare decoction, tablets, granules, pills or oral liquid.

Preferably, the Tibetan medicine compound is prepared into oral liquid.

A preparation method of a Tibetan medicine compound oral liquid comprises the following steps:

(1) weighing radix Berberidis, fructus Hippophae, fructus Chebulae, herba Dracocephali and herba corydalis edulis according to a certain proportion, and cleaning with cold water to remove floating dust;

(2) soaking the medicinal materials obtained in the step (1) in cold water;

(3) adding the soaked medicinal materials in the step (2) into a closed decoction machine, adding water with the volume of 5-10 times of that of the medicinal materials, and decocting the medicinal materials for 2 hours at 115 ℃;

(4) collecting the liquid medicine decocted in the step (3), and filtering four layers of gauze to obtain filtrate;

(5) adding 5-10 times of water, repeating the step (3), decocting with water at 115 deg.C for 2 hr;

(6) collecting the liquid medicine decocted in the step (5), and filtering the liquid medicine with four layers of gauze to obtain filtrate;

(7) mixing the two filtrates obtained in the steps (4) and (6), adding into a rotary evaporator, concentrating the medicinal liquid at 0.05Mpa and 60 deg.C until 1 ml of medicinal liquid is equal to 1g of the original medicinal material;

(8) and (5) adding 0.2% of potassium sorbate into the concentrated liquid medicine in the step (7), canning and sealing.

The Tibetan medicine compound is applied to the preparation of the medicine for treating calf gastrointestinal diseases.

The Tibetan medicine compound is applied to the preparation of the medicine for treating calf colitis.

The Tibetan medicine compound is applied to the preparation of the medicine for treating calf diarrhea.

Compared with the prior art, the Tibetan medicine compound for treating calf diarrhea provided by the invention has the following advantages and remarkable progress:

(1) the preparation is very convenient to use, can be decocted with water, is convenient for animals to take by pouring, stirring and drinking water, is easy to absorb by intestinal mucosa, has the effect of inhibiting or killing escherichia coli pathogenic bacteria causing infection, inhibits intestinal epithelial cell from dropping, reduces mucosal edema, reduces inflammatory cell infiltration, promotes diarrhea to heal quickly, is not easy to generate bacterial drug resistance, and does not influence the production performance of animals.

(2) Has hemostatic and analgesic effects, and also has effects of activating cells, repairing damaged cells, and promoting tissue and cell repair and regeneration.

(3) The product is green and safe without irritant, anesthetic and hormone substances, and the five substances can play a synergistic role in stopping diarrhea, astringing intestines and diminishing inflammation, and can play a highly effective treatment role in calf diarrhea.

Drawings

FIG. 1 weight effect of Tibetan medicine compound on DSS-induced colitis in mice

FIG. 2 influence of Tibetan medicine compound on the index of activity of DSS-induced colitis disease in mice

FIG. 3 Change in MPO content in Colon tissues of mice

FIG. 4 changes in MDA content in the Colon group of mice

FIG. 5 SOD content variation in colon tissue of mice

FIG. 6 changes in GSH content in Colon tissue of mice

Detailed Description

The protection scope of the present invention is described in detail with reference to the following specific examples, but it should be noted that the protection scope of the present invention is not limited to the following examples, and the application of the Tibetan medicine compound in preparing the drug for treating calf diarrhea is protected, and the protection scope includes different dosage forms, dosages, combination drugs, etc. All technical solutions which can be derived from a technical solution by a person skilled in the art through logical analysis, inference and experiment according to the technical solutions of the present invention are within the scope of the claimed invention.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter herein. In this application, it must be noted that, unless the context clearly dictates otherwise, as used in this specification and the claims, the singular forms "a," "an," and "the" include plural referents. It should also be noted that the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the term "comprising" as well as other forms, such as "includes," "including," and "containing," are used without limitation.

The Oxford Cup (Oxford Cup) method described in the following examples is a method for measuring the potency of antibiotics, and can be generally classified into a two-dose method and a three-dose method. The specific operation method comprises the following steps: the sterilized agar medium was heated to completely melt and poured into petri dishes, 15ml each (lower layer), and allowed to solidify. Further, the melted PDA medium was cooled to about 50 ℃ and mixed with the test bacteria, and 5ml of the culture medium mixed with the test bacteria was added to the solidified medium to be solidified (upper layer). Directly and vertically placing an Oxford cup (a round small tube with the inner diameter of 6mm, the outer diameter of 8mm and the height of 10mm, the two ends of the tube are smooth, or a glass tube or a porcelain tube) on the surface of the culture medium in a sterile operation, slightly pressurizing to ensure that the Oxford cup is in contact with the culture medium without a gap, adding a sample to be detected (fermentation liquid) into the Oxford cup, wherein the Oxford cup can be generally filled to 240 microliters and does not overflow. After the mixture is filled, the mixture is cultured for 16 to 18 hours at 37 ℃, and the result is observed, and the bacteriostatic circle can be measured directly by using a ruler. During the culture, on one hand, the test bacteria start to grow, and on the other hand, the antibiotics are diffused in a spherical shape, and the closer to the cup, the higher the antibiotic concentration is, and the farther from the cup, the lower the antibiotic concentration is. As the concentration of the antibiotic is reduced, a minimum inhibitory concentration zone exists, and bacteria can not grow but are in a transparent circle in the zone range, so that the zone is called as an 'inhibitory zone'. The higher the antibiotic concentration, the larger the zone of inhibition.

The gastrointestinal diseases include all gastrointestinal diseases caused by damage to the mucosa of the stomach and duodenum. Gastrointestinal disorders may be caused by a variety of factors such as, but not limited to, alcohol, smoking, stress, drugs, and combinations thereof. The drugs causing gastrointestinal diseases are represented by, but not limited to, non-steroidal anti-inflammatory drugs, steroids, and the like. Representative of the nonsteroidal anti-inflammatory drugs are indomethacin, diclofenac, aspirin, acetaminophen, ibuprofen, meloxicam, naproxen, and the like.

Examples 1 to 5:

the invention provides a preparation method of a Tibetan medicine compound for treating calf diarrhea, which comprises the following steps:

the Tibetan medicine compound formula is shown in table 1, and the data in table 1 are mass ratio.

TABLE 1 Tibetan medicine compound with different proportions

The preparation method of the Tibetan medicine compound oral liquid comprises the following steps:

(1) weighing radix Berberidis, fructus Hippophae, fructus Chebulae, herba Dracocephali and herba corydalis edulis according to a certain proportion, and cleaning with cold water to remove floating dust;

(2) soaking the medicinal materials obtained in the step (1) in cold water for 24 hours;

(3) adding the soaked medicinal materials in the step (2) into a closed decoction machine, adding water with the volume of 5-10 times of that of the medicinal materials, and decocting the medicinal materials for 2 hours at 115 ℃;

(4) collecting the liquid medicine decocted in the step (3), and filtering four layers of gauze to obtain filtrate;

(5) adding 5-10 times of water into the residue, repeating step (3), decocting with water at 115 deg.C for 2 hr;

(6) collecting the liquid medicine decocted in the step (5), and filtering the liquid medicine with four layers of gauze to obtain filtrate;

(7) mixing the two filtrates obtained in the steps (4) and (6), adding into a rotary evaporator, concentrating the medicinal liquid at 0.05Mpa and 60 deg.C until 1 ml of medicinal liquid is equal to 1g of the original medicinal material;

(8) adding 0.2% potassium sorbate, canning and sealing.

Comparative example:

the prescription composition is as follows: 5 parts of myrobalan, 2 parts of dracocephalum cochinchinensis, 2 parts of corydalis tuber and the balance of water.

The preparation process comprises the following steps: weighing myrobalan, dracocephalum tanguticum and corydalis sinica in proportion, washing with cold water to remove floating dust, soaking in cold water for 24 hours, adding 5-10 times of water into a sealed decocting machine, decocting at 115 ℃ for 2 hours, filtering the filtrate by four layers of gauze, adding 5-10 times of water into dregs, decocting at 115 ℃ for 2 hours, filtering the filtrate by four layers of gauze to obtain filtrate, combining the two times of liquid medicine, adding a rotary evaporator, at 0.05Mpa and 60 ℃, concentrating the liquid medicine, concentrating until 1 ml of liquid medicine is equal to 1g of the original medicinal material, adding 0.2% of potassium sorbate, canning and sealing.

Example 6 study on antidiarrheal, analgesic and bacteriostatic effects of Tibetan medicine compound

1. Research on bacteriostatic effect

Taking 200 g of radix berberidis, 300 g of sea buckthorn, 500 g of myrobalan, 200 g of dracocephalum tanguticum and 200 g of corydalis ochotensis in example 1, 300 g of radix berberidis, 30 g of sea buckthorn, 500 g of myrobalan, 200 g of dracocephalum tanguticum and 200 g of corydalis ocharioides in example 2, 200 g of radix berberidis, 400 g of sea buckthorn, 500 g of myrobalan, 200 g of dracocephalum tanguticum and 200 g of corydalis ocharioides in example 3, 100 g of radix berberidis, 300 g of sea buckthorn, 500 g of myrobalan, 200 g of dracocephalum tanguticum and 200 g of corydalis ocharioides in example 4, carrying out bacteriostasis test on high drug-resistant escherichia coli separated from diarrhea calves by an Oxford cup method, cooling the high-pressure sterilized agar culture medium to 50 ℃ and pouring the high-resistant escherichia coli into a 9-MH-diameter one-time culture medium, irradiating the culture medium under an ultraviolet lamp, and then solidifying the culture medium, sealing the sealing film in a refrigerator at 4 deg.C for use. Inoculating the isolated strain of Escherichia coli into a common nutrient broth, and incubating overnight at 37 deg.C for 12-16 h; the nutrient broth culture was adjusted to 0.5 mciro turbidity using a bacterial turbidimeter. And uniformly coating the diluted bacteria liquid on an MH culture medium by using a sterilized cotton swab, so as to ensure that the bacteria liquid is uniformly coated. 4 slightly heated sterilized Oxford cups (inner diameter is 6 +/-0.1 mm, outer diameter is 8 +/-0.1 mm, and height is 10 +/-0.1 mm) are placed on the surface of the MH agar culture medium coated with the bacteria liquid to be in seamless contact with the culture medium, the prepared Tibetan medicine compound oral liquid is diluted into 8 gradient concentrations of 2000mg/mL, 1000mg/mL, 500mg/mL, 250mg/mL, 125mg/mL, 64mg/mL, 32mg/mL and 16mg/mL, the concentration is numbered from high to low in sequence as 1-8, and an Oxford cup method is used for carrying out an escherichia coli bacteriostasis test. Numbering 1-8 from high to low, respectively, adding 300 μ L of the solution into each Oxford cup, and culturing at 37 deg.C for 18-24 h. The vernier caliper measures the size of the zone of inhibition, and the average value is calculated by repeating the measurement for 3 times. The bacterial inhibition diameter is less than 10mm, and the bacterial inhibition is insensitive; the diameter of the bacteriostatic circle is more than or equal to 10mm and less than 15mm, the mild sensitivity is realized, and the diameter of the bacteriostatic circle is more than or equal to 15mm and less than 20mm, the moderate sensitivity is realized; the bacteriostatic diameter is more than 20mm, and the high sensitivity is realized. The experimental results are shown in table 2, the Tibetan medicine compound has a remarkable bacteriostatic effect on escherichia coli, under the conditions that the concentration is 1000mg/mL, 500mg/mL and 250mg/mL, the sensitivity of the escherichia coli to the compound is the same as that of the compound in example 1 and example 3, namely moderate sensitivity, moderate sensitivity and mild sensitivity, and the sensitivity of the escherichia coli to the compound in example 5 is moderate sensitivity, mild sensitivity and mild sensitivity. Examples 2 and 4 had too high inhibitory concentration against e.coli and were therefore eliminated from animal experiments. The Tibetan medicine compound can inhibit escherichia coli to different degrees in example 1, example 3 and example 5.

TABLE 2 inhibition zone diameters for E.coli at different concentrations for different formulations

2. Study of analgesic Effect

The experiment adopts a Tibetan veterinary drug compound pharmacodynamic test, namely an acetic acid induced mouse writhing test to research the analgesic effect of a Tibetan drug compound on a mouse, and the Tibetan drug is prepared according to 200 g of root of Chinese barberry, 300 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucophyllum and 200 g of corydalis tuber in example 1, 200 g of root of Chinese barberry, 400 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucophyllum and 200 g of corydalis tuber in example 3, 300 g of root of Chinese barberry, 100 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucophyllum and 200 g of corydalis tuber in example 5. BALB/c mice were 50, weighing (20. + -.2) g, male and female halves, randomly divided into 5 groups of 10 mice each. The first group was a model group, the second group was an indomethacin positive control group (3mg/kg · bw), the third group was example 1(2mL/kg · bw), the fourth group was example 3(2mL/kg · bw), and the fifth group was example 5(2mL/kg · bw). The mice in the model group are subjected to intragastric administration of 0.4m L/mouse of physiological saline; the positive control group mice are administered with 3mg/kg indometacin by gastric lavage; after the mice in each group are subjected to intragastric administration for 1 hour, 0.6% glacial acetic acid solution 0.2m L is injected into the abdominal cavity of each mouse, the incubation period of writhing of the mice after the glacial acetic acid injection and the writhing frequency within 30min are observed and recorded, and the writhing reaction inhibition rate is calculated.

The inhibition rate is (number of model group writhing times-test group writhing times)/number of model group writhing times multiplied by 100%

As can be seen from table 3, the test groups of examples 1, 3 and 5 all significantly inhibited the number of writhing caused by intraperitoneal injection of glacial acetic acid, and the test groups of examples 1 and 3 had the same inhibition rate as the positive control group, indicating that examples 1 and 3 had significant peripheral analgesic effect.

TABLE 3 comparison of writhing frequency and inhibition ratio of different groups of mice

The pain threshold values of the mice at different time points are detected, the specific results are shown in table 4, and the pain threshold values of the mice in the positive control group, the experimental groups of examples 1, 3 and 5 at the time points of 30min, 60 min and 90min are compared with the pain threshold values of the mice in the model group, so that the difference is not significant (P is more than 0.05). At 120min, compared with the pain threshold of the positive control group mice, the test group mice of examples 1, 3 and 5 and the pain threshold of the model group mice, the difference is significant (P < 0.05). It is demonstrated that examples 1, 3 and 5 have a certain central analgesic effect.

TABLE 4 comparison of pain thresholds at different time points

3. Study of anti-inflammatory Effect

The Tibetan medicine oral liquid is prepared from 200 g of root of Chinese barberry, 300 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 1, 200 g of root of Chinese barberry, 400 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 3, and 300 g of root of Chinese barberry, 100 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 5. BALB/c mice were 50, weighing (20. + -.2) g, male and female halves, randomly divided into 5 groups of 10 mice each. The first group was a model group, the second group was an indomethacin positive control group (3mg/kg · bw), the third group was example 1(2mL/kg · bw), the fourth group was example 3(2mL/kg · bw), and the fifth group was example 5(2mL/kg · bw). Model group mice were given saline, 0.4m L/mouse; the administration of the drug is carried out on the mice for 5 days in advance, the gastric lavage is carried out for 1 time every day, the drug is administered for 1 hour after the last administration, 0.03m L dimethylbenzene is evenly smeared on the front and back surfaces of the right ear of the mouse, and the left ear is used as a control. After 1h, the mice were sacrificed by removing their necks, two round ear pieces were punched along the same portions of the two ears at the base line of the auricle with a punch having a diameter of 8mm, and the swelling inhibition rate was calculated by weighing.

Swelling degree-right ear weight-left ear weight; swelling rate ═ swelling degree/left ear weight × 100%

The swelling inhibition rate is (swelling degree of model group-swelling degree of administration group)/swelling degree of model group x 100%

The results of the xylene-induced ear swelling test of mice (as shown in table 5) show that examples 1, 3 and 5 have certain inhibitory effect on the xylene-induced ear swelling of mice. The swelling rate of the mice in example 1 is 54.90%, the inhibition rate is 30.66%, and the difference is very obvious compared with the swelling rate of the mice in a model group (P < 0.01); example 3 the mice had 56.33% swelling rate and 30.0% inhibition rate, and the difference was significant compared with the model group (P < 0.05).

TABLE 5 comparison of ear swelling Rate in different groups of mice

4. Intestinal propulsion experiment

The Tibetan medicine oral liquid is prepared from 200 g of root of Chinese barberry, 300 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 1, 200 g of root of Chinese barberry, 400 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 3, and 300 g of root of Chinese barberry, 100 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 5. BALB/c mice were 50, weighing (20. + -.2) g, male and female halves, randomly divided into 5 groups of 10 mice each. The first group is a model group, the second group is a compound diphenoxylate positive control group (3mg/kg · bw), the third group is example 1(2mL/kg · bw), the fourth group is example 3(2mL/kg · bw), and the fifth group is example 5(2mL/kg · bw). The mice in the model group are gavaged with physiological saline, 0.4m L/mouse; the positive control group, mice in examples 1, 3 and 5 were gavaged once a day for 5 days. Fasting before the last administration for 12h, and after the last administration for 1h, performing intragastric administration on the 5.0% carbon powder suspension to mice according to the ratio of 0.15m L/mouse, performing intragastric administration for 30min, performing neck dislocation, opening abdominal cavity to separate mesentery, cutting the intestinal canal from the upper end to pylorus and from the lower end to ileocecum, and placing on a tray. The intestinal tube was gently straightened and the length of the intestinal tube was measured as the "total length of small intestine". The distance from the pylorus to the leading edge of the ink serves as the "distance the ink advances in the intestinal tract". Percent ink propulsion was calculated by formula and care was taken to see if the small bowel volume increased in each group.

Percent (%) carbon powder propulsion (distance from the anterior segment of the carbon powder to the pylorus/total length of small intestine X100%)

The results of the carbon ink propulsion test (as shown in table 6) show that the intestinal propulsion rate of the mice in example 5 is 62.82%, and compared with the intestinal propulsion rate of the mice in the model group, the intestinal propulsion rate is remarkably reduced (P < 0.05); the intestinal propulsion rate of the mice of examples 1 and 3 and the positive control group can be reduced very significantly (P <0.01) compared with the model group. The results show that the examples 1 and 3 can obviously reduce carbon ink propelling, have a certain antidiarrheal function and have the effect similar to that of the compound diphenoxylate.

TABLE 6 comparison of intestinal propulsive Effect in different groups of mice

5. Senna leaf diarrhea test

The Tibetan medicine oral liquid is prepared from 200 g of root of Chinese barberry, 300 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 1, 200 g of root of Chinese barberry, 400 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 3, and 300 g of root of Chinese barberry, 100 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 5. BALB/c mice were 50, weighing (20. + -.2) g, male and female halves, randomly divided into 5 groups of 10 mice each. The first group is a model group, the second group is a compound diphenoxylate positive control group (3mg/kg · bw), the third group is example 1(2mL/kg · bw), the fourth group is example 3(2mL/kg · bw), and the fifth group is example 5(2mL/kg · bw). The mice in the model group are gavaged with physiological saline, 0.4m L/mouse; the positive control group, mice in examples 1, 3 and 5 were gavaged once a day for 5 days. Before the last administration, fasting is not prohibited for 12 hours, and after the last administration for 0.5 hour, the intragastric concentration is 1g/m L senna leaf and 0.4m L/patient. And observing each mouse in a single cage, paving a water-absorbing filter paper under each cage, changing the filter paper every 2 hours, and calculating the diarrhea index of the mouse at 2, 4 and 6 hours respectively.

The diarrhea index is the rate of loose stool x the level of loose stool x 100%

The results show that the diarrhea phenomenon of the model group mice is obviously enhanced after the administration of the senna leaves, the diarrhea caused by the senna leaves can be greatly reduced within 2-4h in example 1, the diarrhea index of the compound diphenoxylate group mice is 1.83, the difference is obvious compared with the model group (P <0.05), the diarrhea index of the example 1 mice is 1.75, and the difference is very obvious compared with the model group (P < 0.01).

TABLE 7 diarrhea index comparison of different groups of mice

6. Treatment effect of Tibetan medicine compound on ulcerative colitis

The Tibetan medicine oral liquid is prepared from 200 g of root of Chinese barberry, 300 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 1, 200 g of root of Chinese barberry, 400 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 3, and 300 g of root of Chinese barberry, 100 g of sea buckthorn, 500 g of myrobalan, 200 g of glaucocalyx canadensis and 200 g of corydalis tuber in example 5. SPF grade BALB/c mice were taken as 50 mice, randomly divided into 5 groups of 10 mice each, and were acclimatized for one week for testing. Respectively as normal control group, model group, sulfasalazine positive control group, compound 1, 2, and 3 groups. The mice in the drug group were administered 3 days in advance, and the sulfasalazine positive control group (2mg/kg · bw), example 1(2mL/kg · bw), example 3(2mL/kg · bw), example 5(2mL/kg · bw); the model group and control group mice were given the same volume of distilled water daily. Except for the normal control group, each group of mice freely drunk DSS water with a concentration of 3.5% for 6 days continuously. And observing the characteristics of the excrement of the mouse every day, detecting the occult blood of the excrement, and weighing the mouse. At the end of the experiment, the colon was isolated and frozen at-80 ℃.

The treatment result of the Tibetan medicine compound on the mouse ulcerative colitis model shows that the weight of the model group mice induced by the DSS begins to lose and the spirit is low in the 2 nd day of the test; the phenomena of soft stool and diarrhea begin to appear at the 3 rd time of the test, and the stool occult blood is detected as blue; the 4d mouse has reduced activity, is coiled in a mouse cage, is sleepy, begins to have bloody stool, has obvious weight reduction and obviously increases the disease activity index (P < 0.01); at 3d, the mice all have bloody stools, the body weight of the mice is extremely different from that of the normal control group mice (P <0.01), and the disease activity index is extremely increased (P < 0.01). Example 3 the mice in the intervention group had good mental activities, the individual mice had the phenomena of soft stool and diarrhea, no bloody stool and slightly increased body weight, and the DAI score of the 4d mice from the experiment was significantly lower than that of the model group (P < 0.01). Example 1 mice in the intervention group had slightly reduced body weight, mice in the 4d experiment had greatly different body weight from those in the normal control group (P <0.01), poor mental activity and diarrhea, DAI scores in the 5d experiment were significantly different from those in the model group (P <0.01) and differences in the 6d experiment (P < 0.05). In contrast, when the mice are intervened by the method of example 5, the weight loss of the mice is obvious, and the weight loss of the mice is slightly improved compared with that of a model group, but the difference is not significant (P is more than 0.05). It can be seen that example 3 has the best therapeutic effect on colitis in mice (see fig. 1 and 2).

The detection result of Myeloperoxidase (MPO) in colon tissues shows that the MPO activity in the colon tissues of the model mice is obviously improved (P <0.01) compared with that of a normal control group (see figure 3); the MPO activity in the colon tissues of mice in the administration group is obviously reduced compared with that in the model group, and the MPO activity reduction of the examples 1 and 3 is extremely obvious (P <0.01) compared with that in the model group; example 5 was significantly different compared to the model group (P < 0.05); it is demonstrated that examples 1, 3, 5 have the effect of reducing MPO activity in colitis tissue.

The measurement results of the content of Malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and the activity of glutathione peroxidase (GSH-Px) show that the content of MDA in colon tissues of the mice in the model group is remarkably increased (P is less than 0.01) compared with that in a normal control group. Examples 1, 5 significantly reduced the amount of MDA in colon tissue (P <0.01) compared to the model group (see fig. 4); it is demonstrated that examples 1, 5 have the effect of reducing the MDA content in colitis tissues to normal levels. In the DSS-induced mouse colitis model, the activity of SOD, GSH in colon tissue was reduced. Compared with the model group, the activity of SOD and GSH in colon tissues of mice in example 3 is increased and has a very significant difference (P <0.01), and the activity of SOD and GSH in colitis tissues of positive control groups and example 5 groups is significantly different (P <0.05) (see figures 5 and 6), which indicates that examples 1 and 3 have the function of increasing the activity of SOD and GSH in colitis tissues.

A large number of experiments prove that the Tibetan medicine compound for treating calf diarrhea has the following advantages:

(1) has effects in relieving diarrhea and astringing intestine. The Tibetan medicine compound of the invention mainly comprises root of Chinese barberry, sea backthern, myrobalan, dracocephalum tanguticum and corydalis tuber. Radix Berberidis has the effects of clearing heat, eliminating dampness, purging pathogenic fire and removing toxic substances; the sea buckthorn has the effects of promoting digestion, removing stagnation, promoting blood circulation and removing blood stasis; the fructus Chebulae has effects of relieving diarrhea with astringents, astringing lung and relieving cough; the dracocephalum tanguticum has the effects of harmonizing stomach, soothing liver, clearing heat and promoting diuresis; the corydalis tuber has the effects of clearing heat and diminishing inflammation. The Tibetan medicine compound preparation can be used for remarkably reducing the intestinal propulsion rate of mice, remarkably reducing the diarrhea index and not influencing the animal production performance.

(2) Has antibacterial and antiinflammatory effects. The Tibetan medicine compound decoction has strong bacteriostatic and bactericidal effects on high-drug-resistance Escherichia coli isolates of diarrhea calves, and is not easy to generate bacterial drug resistance.

(3) Promoting diarrhea rapid healing, inhibiting intestinal epithelial cell shedding, reducing mucosal edema, reducing inflammatory cell infiltration, reducing organism peroxidase content, increasing superoxide dismutase content, relieving intestinal mucosa injury, and protecting intestinal mucosa.

(4) Has obvious peripheral analgesic and central analgesic effects.