CN112618600B - Tibetan medicine compound for treating calf gastrointestinal diseases and preparation method thereof - Google Patents
Tibetan medicine compound for treating calf gastrointestinal diseases and preparation method thereof Download PDFInfo
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- CN112618600B CN112618600B CN202110041339.6A CN202110041339A CN112618600B CN 112618600 B CN112618600 B CN 112618600B CN 202110041339 A CN202110041339 A CN 202110041339A CN 112618600 B CN112618600 B CN 112618600B
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Abstract
The invention relates to a Tibetan medicine compound for treating calf gastrointestinal diseases and a preparation method thereof, and relates to the technical field of medicinal preparations. The Tibetan medicine compound is prepared from the following raw materials in parts by weight: 1-3 parts of radix berberidis, 1-3 parts of herba elsholtziae, 1-5 parts of gentiana macrophylla, 2 parts of myrobalan, 1 part of dracocephalum tanguticum and 1 part 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.
Description
Technical Field
The invention relates to the technical field of medicinal preparations, in particular to a Tibetan medicine compound for treating calf gastrointestinal diseases and a preparation method 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). 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, management and the like. 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 influences the growth and development of calves, but also influences the renewal of cattle herds and the benign development of the dairy cattle 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 areas. 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 occurs, antibiotics such as gentamicin, norfloxacin, terramycin, norfloxacin and the like are mostly used for treating the disease, but the disease causes are not necessarily caused by bacteria, so that the pertinence is not strong, the problems of easy occurrence of bacterial drug resistance, excessive corresponding animal product drug residues and the like are caused, and the problems of serious public health safety and the like are caused. The calf diarrhea is caused by a plurality of causes, including infectious factors and non-infectious factors.
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. The treatment effect on the complex calf diarrhea still cannot be achieved.
Aiming at the technical problems, the invention provides a Tibetan medicine compound capable of treating calf gastrointestinal diseases aiming at the existing calf gastrointestinal diseases, and the Tibetan medicine compound can quickly stop diarrhea, reduce the use of antibiotics and reduce the generation of bacterial drug resistance.
Disclosure of Invention
The detailed technical scheme of the invention 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, 1-3 parts of herba elsholtziae, 1-5 parts of gentiana macrophylla, 2 parts of myrobalan, 1 part of dracocephalum tanguticum and 1 part 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 herba elsholtziae, 2 parts of gentiana macrophylla, 2 parts of myrobalan, 1 part of dracocephalum tanguticum and 1 part of corydalis tuber.
Preferably, the Tibetan medicine compound is prepared from the following raw materials in parts by weight: 1 part of root of Chinese barberry, 1 part of Chinese mosla herb, 1 part of gentiana macrophylla, 2 parts of myrobalan, 1 part of dracocephalum tanguticum and 1 part of corydalis petiolus.
Preferably, the Tibetan medicine compound is prepared from the following raw materials in parts by weight: 3 parts of root of Chinese barberry, 1 part of herba elsholtziae, 5 parts of gentiana macrophylla, 2 parts of myrobalan, 1 part of dracocephalum tanguticum and 1 part of corydalis tuber.
Preferably, the Tibetan medicine compound is added with pharmaceutically acceptable auxiliary materials to prepare decoction, tablets, granules, pills or oral liquid.
The preparation method of the Tibetan medicine compound oral liquid comprises the following steps:
(1) weighing radix Berberidis, herba Moslae, radix Gentianae Marcrophyllae, fructus Chebulae, herba Dracocephali and herba corydalis Decumbentis in 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) filtering and collecting the liquid medicine decocted in the step (3) by gauze;
(5) filtering the obtained residue, adding 5-10 times of water, repeating step (3), decocting with water at 115 deg.C for 2 hr;
(6) filtering and collecting the decocted liquid medicine in the step (5) by gauze;
(7) combining the two liquid medicines obtained in the step (4) and the step (6), adding the liquid medicines into a rotary evaporator, concentrating the liquid medicines at the temperature of 60 ℃ under the pressure of 0.05Mpa until 1 ml of the liquid medicines are equal to 1g of the original medicinal materials;
(8) and (5) adding 0.2% of potassium sorbate into the concentrated liquid medicine in the step (7), canning and sealing.
The invention also provides application of the Tibetan medicine compound in preparing a medicine for treating calf gastrointestinal diseases.
The invention also provides application of the Tibetan medicine compound in preparing a medicine for treating colitis of calves.
The invention also provides application of the Tibetan medicine compound in preparing a medicine for treating diarrhea type colitis of calves.
The invention also provides application of the Tibetan medicine compound in preparing a medicine for treating calf diarrhea.
Compared with the prior art, the Tibetan medicine compound oral liquid for treating calf diarrhea has the following advantages and remarkable progress:
(1) the oral liquid is very convenient to use, is convenient for animals to take by drenching, mixing with feed and drinking water for administration, is easy to absorb by intestinal mucosa, has the functions of inhibiting or killing escherichia coli pathogenic bacteria causing infection, inhibiting intestinal epithelial cell shedding, reducing mucosal edema, reducing inflammatory cell infiltration, promoting 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 irritative, anesthetic and hormone substances, and the five substances can play a synergistic role in diarrhea relief, intestine astringency and inflammation diminishing, and can play a highly effective treatment role in calf diarrhea.
Drawings
FIG. 1 shows the weight effect of Tibetan medicine compound oral liquid on DSS-induced colitis in mice
FIG. 2 is the effect of compound oral liquid of Tibetan medicine 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 one method for measuring antibiotic potency, and can be generally divided 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.
It should be noted that the following examples are all conventional ones unless otherwise specified.
Examples 1 to 5:
the invention provides a preparation method of a Tibetan medicine compound oral liquid for treating diarrhea-type colitis of calves, which comprises the following steps:
the composition of the Tibetan medicine compound oral liquid is shown in the table, and the data in the table 1 are mass percent.
TABLE 1 Tibetan medicine compound with different proportions
The preparation method comprises the following steps:
(1) weighing radix Berberidis, herba Moslae, radix Gentianae Marcrophyllae, fructus Chebulae, herba Dracocephali and herba corydalis Decumbentis according to a certain proportion, and cleaning with cold water to remove floating dust;
(2) adding the medicinal materials in the step (1), and soaking 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) filtering and collecting the decocted liquid medicine in the step (3) by four layers of gauze;
(5) adding 5-10 times of water into the residue, repeating step (3), decocting with water at 115 deg.C for 2 hr;
(6) filtering four layers of gauze and collecting the decocted liquid medicine in the step (5);
(7) combining the two liquid medicines obtained in the step (4) and the step (6), adding the liquid medicines into a rotary evaporator, concentrating the liquid medicines at the temperature of 60 ℃ under the pressure of 0.05Mpa until 1 ml of the liquid medicines are equal to 1g of the original medicinal materials;
(8) adding 0.2% potassium sorbate, canning and sealing.
Example screening: the compound oral liquid is prepared according to the proportion of the examples 1-5, and the prepared Tibetan medicine compound oral liquid is subjected to related experiments on calf gastrointestinal tract diseases, and the experimental results show that the effects of the examples 1, 3 and 5 are better, and the effects of the examples 2 and 4 are general, so that the following examples only list the results of the examples 1, 3 and 5, but not the results of the examples 2 and 4.
Comparative example:
the prescription composition is as follows: 2 parts of myrobalan, 1 part of dracocephalum tanguticum, 1 part of corydalis petiolata and the balance of water.
The preparation process comprises the following steps: weighing myrobalan, dracocephalum tanguticum and corydalis sinica in proportion, cleaning 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 with water at 115 ℃ for 2 hours, filtering with four layers of gauze, adding 5-10 times of water, decocting with water at 115 ℃ for 2 hours, filtering with four layers of gauze, combining the two liquid medicines, adding a rotary evaporator, concentrating at 0.05Mpa and 60 ℃, concentrating the liquid medicine until 1 ml of liquid medicine is equal to 1 gram of the original medicinal material, adding 0.2% potassium sorbate, canning and sealing.
Example 6: research on antidiarrheal, analgesic and antibacterial effects of Tibetan medicine compound
1. Research on bacteriostatic effect
And performing an antibacterial test of the Tibetan medicine compound oral liquid on the high-drug-resistance Escherichia coli isolate of the diarrhea calves by an oxford cup method. Taking the medicines according to the proportion of the examples 1, 2, 3, 4 and 5, wherein the medicines taken in the example 1 are as follows: 200 g of radix berberidis, 300 g of elsholtzia herb, 200 g of gentiana macrophylla, 200 g of myrobalan, 100 g of dracocephalum tanguticum and 100 g of corydalis tuber, and the medicines taken in the embodiment 2 are as follows: 300 g of root of Chinese barberry, 300 g of elsholtzia, 300 g of gentiana macrophylla, 200 g of myrobalan, 100 g of dracocephalum tanguticum and 100 g of corydalis tuber, and the medicines taken in the embodiment 3 are: 100 g of root of Chinese barberry, 100 g of elsholtzia herb, 100 g of gentiana macrophylla, 200 g of myrobalan, 100 g of dracocephalum tanguticum and 100 g of corydalis tuber, and the medicines taken in the embodiment 4 are as follows: 100 g of root of Chinese barberry, 300 g of elsholtzia, 500 g of gentiana macrophylla, 200 g of myrobalan, 100 g of dracocephalum tanguticum and 100 g of corydalis tuber, and the medicines taken in the embodiment 5 are: 300 g of root of Chinese barberry, 100 g of Chinese mosla herb, 500 g of large-leaved gentian, 200 g of myrobalan, 100 g of dracocephalum tanguticum and 100 g of corydalis tuber.
Pouring MH agar culture medium cooled to 50 ℃ after autoclaving into disposable plastic culture medium with the diameter of 9mm, sealing the sealing film after the culture medium is solidified under the irradiation of an ultraviolet lamp, and placing the sealed film in a refrigerator at 4 ℃ for later 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 1790mg/mL, 1000mg/mL, 500mg/mL, 250mg/mL, 125mg/mL, 62.5mg/mL, 31.25mg/mL and 15.625mg/mL respectively, the concentrations are numbered from high to low in sequence as 1-7, No. 8 is a sterile water control group, 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 results show (table 2) that the compound Tibetan medicines in examples 1, 3 and 5 have bacteriostatic action on escherichia coli, under the conditions of the concentrations of 1790mg/m L, 1000mg/m L and 500mg/m L, the sensitivity of the escherichia coli to the sensitivity conditions in example 1 and example 3 is respectively moderate sensitivity, moderate sensitivity and mild sensitivity, and the sensitivity of the escherichia coli to the sensitivity condition in example 5 is respectively 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 of different formulations for E.coli at different concentrations
2. Study of analgesic Effect
Firstly, it should be noted that the experiment using the compound oral liquid is consistent with the experiment of bacteriostasis effect.
Tibetan veterinary medicine compound pharmacodynamic test-acetic acid induced writhing test of mice, BALB/c mice 50, weight (20 +/-2) g, sex half each, divided into 5 groups at random, each group 10. 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 indomethacin of 3 mg/kg-bw 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, which indicates that examples 1 and 3 had significant peripheral analgesic effect (see table 3).
TABLE 3 comparison of writhing frequency and inhibition ratio of different groups of mice
The test results show that the pain threshold values of the positive control group and the test group mice of the examples 1, 3 and 5 at the time points of 30min, 60 min and 90min are compared with the pain threshold value of the model group mice, and the difference is not significant (P > 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 shown that examples 1 and 3 have a certain central analgesic effect (see Table 4).
TABLE 4 comparison of pain thresholds at different time points
3. Study of anti-inflammatory Effect
Firstly, it should be noted that the experiment adopts the compound oral liquid which is consistent with the experiment of the bacteriostatic effect.
BALB/c mice were 50 mice (20. + -.2 g, half each female) and half each male, 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 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 and 3 have certain inhibitory effect on the xylene-induced ear swelling of mice. The swelling rate of the mice of the examples 3 and 5 is 53.15 percent and 52.78 percent, the inhibition rate is 32.46 percent and 34.02 percent, and the difference is very obvious compared with the swelling rate of the mice of a model group (P < 0.01).
TABLE 5 comparison of ear swelling Rate in different groups of mice
4. Intestinal propulsion experiment
Firstly, it should be noted that the experiment using the compound oral liquid is consistent with the experiment of bacteriostasis effect.
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 mice in the positive control group, examples 1, 3 and 5 groups 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 carbon ink propulsion test results (as shown in table 6) show that the intestinal propulsion rates of the mice of the examples 1 and 3 are 50.46 percent and 52.06 percent, and the intestinal propulsion rate is extremely reduced compared with the intestinal propulsion rate of the mice of the model group (P < 0.01); example 5 and the intestinal propulsion rate of the positive control group mice are compared with the model group, the intestinal propulsion rate of the mice can be obviously reduced (P <0.05), and the compound diphenoxylate antidiarrheal compound has a certain antidiarrheal function and the effect is similar to that of the compound diphenoxylate.
TABLE 6 comparison of intestinal Propulsion Effect in different groups of mice
5. Senna leaf diarrhea test
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 mice in the model group is obviously enhanced after the administration of the senna leaves, the diarrhea index of the mice in the compound diphenoxylate group is 1.83 within 2-4h, the difference is obvious compared with the model group (P <0.05), the diarrhea index of the mice in the example 1 is 1.93, the difference is extremely obvious compared with the model group (P <0.01), the diarrhea caused by the senna leaves can be greatly reduced within 2-4h in the example 1, and the effect is similar to that of the compound diphenoxylate.
TABLE 7 diarrhea index comparison of different groups of mice
6. Treatment effect of Tibetan medicine compound on ulcerative colitis
Firstly, it should be noted that the experiment using the compound oral liquid is consistent with the experiment of bacteriostasis effect.
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. The control group is a normal control group, a model group, a sulfasalazine positive control group, and examples 1, 3 and 5. The mice in the drug group were administered 3 days in advance, and the drugs in the corresponding doses were administered daily in accordance with sulfasalazine positive control group (2mg/kg · bw), example 1(2mL/kg · bw), example 3(2mL/kg · bw), and 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 appear at the 3 rd time of the test, and the stool occult blood is detected to be 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 1 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 3 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 1 has the best therapeutic effect on colitis in mice (see fig. 1 and 2).
The result of Myeloperoxidase (MPO) detection in colon tissue shows (see figure 3), compared with the normal control group, the MPO activity in the colon tissue of the model group mice is obviously increased (P < 0.01); 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, 3 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 and 3 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 oral liquid for treating the calf diarrhea type colitis 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, herba elsholtziae, gentiana macrophylla, myrobalan, dracocephalum tanguticum and corydalis edulis. Radix Berberidis has the effects of clearing heat, eliminating dampness, purging pathogenic fire and removing toxic substances; herba Moslae has effects of dispelling pathogenic wind heat; the gentiana macrophylla has the effects of expelling wind-damp and clearing damp-heat; 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 and remarkably reducing the diarrhea index.
(2) Has antibacterial and antiinflammatory effects. The Tibetan medicine compound oral liquid has strong bacteriostatic and bactericidal effects on high-drug-resistance escherichia coli isolates of diarrhea calves.
(3) Reducing the content of peroxidase in organism, increasing the content of superoxide dismutase, relieving intestinal mucosa injury, and protecting intestinal mucosa.
(4) Has obvious peripheral analgesic and central analgesic effects.
Claims (10)
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, 1-3 parts of herba elsholtziae, 1-5 parts of gentiana macrophylla, 2 parts of myrobalan, 1 part of dracocephalum tanguticum and 1 part of corydalis tuber.
2. The Tibetan medicine compound of claim 1, which is prepared from the following raw materials in parts by weight: 2 parts of radix berberidis, 3 parts of herba elsholtziae, 2 parts of gentiana macrophylla, 2 parts of myrobalan, 1 part of dracocephalum tanguticum and 1 part of corydalis tuber.
3. The Tibetan medicine compound of claim 1, which is prepared from the following raw materials in parts by weight: 1 part of radix berberidis, 1 part of herba elsholtziae, 1 part of radix gentianae macrophyllae, 2 parts of myrobalan, 1 part of dracocephalum tanguticum and 1 part of corydalis tuber.
4. The Tibetan medicine compound of claim 1, which is prepared from the following raw materials in parts by weight: 3 parts of root of Chinese barberry, 1 part of herba elsholtziae, 5 parts of gentiana macrophylla, 2 parts of myrobalan, 1 part of dracocephalum tanguticum and 1 part 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 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, herba Moslae, radix Gentianae Marcrophyllae, fructus Chebulae, herba Dracocephali and herba corydalis Decumbentis in 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) filtering and collecting the liquid medicine decocted in the step (3) by gauze;
(5) filtering the obtained residue, adding 5-10 times of water, repeating step (3), decocting with water at 115 deg.C for 2 hr;
(6) filtering and collecting the decocted liquid medicine in the step (5) by gauze;
(7) combining the two liquid medicines obtained in the step (4) and the step (6), adding the liquid medicines into a rotary evaporator, concentrating the liquid medicines at the temperature of 60 ℃ under the pressure of 0.05Mpa until 1 ml of the liquid medicines are equal to 1g of the original medicinal materials;
(8) and (5) adding 0.2% of potassium sorbate into the concentrated liquid medicine in the step (7), canning and sealing.
7. 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.
8. Use of the Tibetan medicine compound of any one of claims 1-4 in the preparation of a medicament for treating colitis in calves.
9. Use of the Tibetan medicine compound of any one of claims 1-4 in the preparation of a medicament for treating diarrhea-type 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.
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