Application of traditional Chinese medicine composition in preparation of medicine for preventing and treating bacterial infectious diarrhea

Abstract

The invention relates to the technical field of traditional Chinese medicines, in particular to application of a traditional Chinese medicine composition in preparation of a medicine for preventing and treating bacterial infectious diarrhea. The traditional Chinese medicine composition is prepared from 1 to 100 parts of cortex magnoliae officinalis, 1 to 100 parts of scorched betel nut, 1 to 100 parts of roasted fructus tsaoko, 1 to 100 parts of herba ephedrae, 1 to 100 parts of semen armeniacae amarae, 1 to 100 parts of rhizoma seu radix notopterygii, 1 to 100 parts of rhizoma zingiberis recens, 1 to 100 parts of herba pogostemonis, 1 to 100 parts of herba eupatorii, 1 to 100 parts of rhizoma atractylodis, 1 to 160 parts of poria cocos, 1 to 120 parts of rhizoma atractylodis macrocephalae, 1 to 100 parts of gypsum, 1 to 100 parts of scorched fructus crataegi, 1 to 100 parts of scorched medicated leaven, 1 to 100 parts of scorched malt, 1 to 100 parts of earthworm, 1 to 100 parts of radix The traditional Chinese medicine composition can reduce the level of inflammatory factors in the body and relieve intestinal infiltration of inflammatory cells, so that the anti-diarrhea effect is achieved.

Description

[0001] This application claims the priority of the Chinese patent application filed with the China Patent Office on June 14, 2023, with application number 202310706426.8 and invention name "Application of Chinese herbal medicine composition in the preparation of drugs for preventing and treating bacterial infectious diarrhea", the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The invention relates to the technical field of traditional Chinese medicines, and in particular to application of a traditional Chinese medicine composition in preparing medicines for preventing and treating bacterial infectious diarrhea. Background Art

[0003] The pathogenic causes of animal diarrhea are divided into two categories: non-pathogenic factors (feeding environment, feeding methods, management, etc.) and pathogenic factors (bacteria, viruses, parasites, etc.). Bacterial infectious diarrhea refers to a group of common intestinal infectious diseases caused by various bacteria, with diarrhea as the main manifestation. Among them, Escherichia coli is one of the important pathogens causing diarrhea. Bacterial pathogenic diarrhea caused by pathogenic Escherichia coli is characterized by rapid onset, high mortality, and strong infectivity.

[0004] There are many kinds of anti-diarrhea drugs on the market, mainly Western medicines, and commonly used antibacterial drugs include berberine, gentamicin, ciprofloxacin, norfloxacin (i.e. quinolone drugs), etc. Western medicine has certain disadvantages in treating diarrhea, that is, Western medicine has many side effects and even addiction. Abuse of antibiotics can easily cause side effects of varying degrees, such as nausea, vomiting, loss of appetite, and even affect the liver, kidney and hematopoietic function.

[0005] Chinese medicine is a traditional Chinese medicine with extremely rich sources, which can be derived from animals, plants and minerals, and contains a variety of biologically effective ingredients. Most of them are made from natural plants, which are rich in resources, easy to obtain, and have little toxic side effects. Therefore, providing a related product developed based on Chinese herbal medicine has broad application value in the prevention and treatment of animal diarrhea. Summary of the invention

[0006] In view of this, the present invention provides the use of a Chinese medicine composition in the preparation of a drug for preventing and treating bacterial infectious diarrhea. The present invention has found through experiments that the Chinese medicine composition granules of the present invention can regulate cytokine levels, and the Chinese medicine composition granules of the present invention can effectively reduce intestinal inflammation and protect intestinal tissue.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides the use of a Chinese medicine composition in preparing a medicine for treating and / or preventing bacterial infectious diarrhea;

[0009] The Chinese medicine composition comprises: 1-100 parts of Magnolia officinalis, 1-100 parts of burnt Areca catechu, 1-100 parts of stewed Tsaoko fruit, 1-100 parts of Ephedra, 1-100 parts of Bitter Apricot, 1-100 parts of Notopterygium wilfordii, 1-100 parts of Ginger, 1-100 parts of Patchouli, 1-100 parts of Cyperus rotundus, 1-100 parts of Atractylodes macrocephala, 1-160 parts of Poria, 1-120 parts of Atractylodes macrocephala, 1-100 parts of Gypsum, 1-100 parts of burnt Crataegus pinnatifida, 1-100 parts of burnt Liu Shenqu, 1-100 parts of burnt malt, 1-100 parts of Pheretima, 1-100 parts of Cynanchum cyrtonema, 1-100 parts of Cynomorium wilfordii and / or 1-100 parts of Lepidium chinense.

[0010] In some specific embodiments of the present invention, the treatment and / or prevention of bacterial infectious diarrhea includes any of the following:

[0011] (I) Improve fever; and / or

[0012] (II), improve abdominal pain; and / or

[0013] (III) Improve diarrhea; and / or

[0014] (IV), improve vomiting; and / or

[0015] (V) Reduce intestinal inflammation.

[0016] In some specific embodiments of the present invention, reducing intestinal inflammation comprises regulating cytokine levels and / or reducing inflammatory cell infiltration.

[0017] In some specific embodiments of the present invention, regulating cytokine levels comprises any of the following:

[0018] (I), reducing the level of IL-6; and / or

[0019] (II), reducing the content of TNF-α; and / or

[0020] (III), reducing the content of IFN-γ; and / or

[0021] (IV) Reduce the content of IL-10.

[0022] In some specific embodiments of the present invention, reducing inflammatory cell infiltration includes reducing the MPO activity level in intestinal tissue.

[0023] In some specific embodiments of the present invention, the traditional Chinese medicine composition includes: 10-80 parts of Magnolia officinalis, 10-80 parts of charred Areca nut, 10-80 parts of stewed grass fruit, 10-60 parts of Ephedra, 10-60 parts of bitter almond, 10-80 parts of Notopterygium, 10-60 parts of ginger, 10-80 parts of Patchouli, 10-60 parts of Cyperus rotundus, 10-80 parts of Atractylodes, 10-160 parts of Poria, 10-120 parts of Atractylodes, 10-80 parts of Gypsum, 10-50 parts of charred Crataegus pinnatifida, 10-80 parts of charred Liu Shenqu, 10-60 parts of charred malt, 10-80 parts of Pheretima, 10-80 parts of Cynomorium wilfordii, 10-60 parts of Cyperus rotundus and / or 10-80 parts of Lemongrass.

[0024] In some specific embodiments of the present invention, the Chinese medicine composition comprises: 30-50 parts of Magnolia officinalis, 20-30 parts of charred Areca nut, 30-50 parts of stewed grass fruit, 20-30 parts of Ephedra, 20-30 parts of Bitter Apricot, 30-50 parts of Notopterygium, 30-50 parts of Ginger, 30-50 parts of Patchouli, 20-30 parts of Perilla frutescens, 30-50 parts of Atractylodes, 120-150 parts of Poria, 80-100 parts of Atractylodes macrocephala, 30-50 parts of Gypsum, 20-30 parts of charred Crataegus pinnatifida, 30-50 parts of charred Six-spirited Song, 20-30 parts of charred Malt, 30-50 parts of Pheretima, 30-50 parts of Cynanchum cyrtonema, 20-30 parts of Mianma Cynomorium and / or 30-50 parts of Lemongrass seeds, or a combination of two or more thereof;

[0025] Preferentially, one or a combination of two or more of 50 parts of Magnolia officinalis, 30 parts of scorched Areca nut, 30 parts of stewed Tsaoko fruit, 20 parts of Ephedra, 30 parts of Bitter Apricot, 50 parts of Notopterygium wilfordii, 50 parts of Ginger, 50 parts of Patchouli, 30 parts of Perilla frutescens, 50 parts of Atractylodes macrocephala, 150 parts of Poria, 100 parts of Atractylodes macrocephala, 50 parts of Gypsum, 30 parts of scorched Crataegus pinnatifida, 30 parts of scorched Liu Shenqu, 30 parts of scorched malt, 50 parts of Pheretima, 50 parts of Cynanchum chinense, 30 parts of Cynomorium mongolicum and / or 50 parts of Semen Lepidium.

[0026] In some specific embodiments of the present invention, the Chinese medicine composition includes but is not limited to raw materials, powders, slices or other forms made from raw materials, and extracts, fermentation broths or other forms with the same or similar biological activities.

[0027] In some specific embodiments of the present invention, the preparation method of the traditional Chinese medicine composition comprises: taking a formula amount of one or a combination of two or more of Magnolia officinalis, charred Areca nut, stewed grass fruit, ephedra, bitter almond, Notopterygium wilfordii, ginger, patchouli, Perilla frutescens, Atractylodes lancea, Poria cocos, Atractylodes macrocephala, gypsum, charred hawthorn, charred Liushenqu, charred malt, earthworm, Cynanchum chinense, Mianma Guanzhong and / or Lemongrass seed, reflux extraction, filtering, concentrating, centrifuging, drying, and crushing to obtain an intermediate composition;

[0028] Taking the intermediate composition and mixing it with pharmaceutically acceptable excipients to prepare the Chinese medicine composition;

[0029] The reflux extraction includes: adding 6 times of water for the first time and extracting for 1.5 hours, and adding 4 times of water for the second time and extracting for 1.0 hour.

[0030] In some specific embodiments of the present invention, the acceptable excipients include one or more of a filler, a disintegrant, a lubricant, a suspending agent, a binder, a sweetener, a flavoring agent, a preservative or a matrix.

[0031] In some specific embodiments of the present invention, the preparation method of the traditional Chinese medicine composition comprises: taking 50 parts of Magnolia officinalis, 30 parts of burnt betel nut, 30 parts of stewed tsaoko, 20 parts of ephedra, 30 parts of bitter almond, 50 parts of notopterygium, 50 parts of ginger, 50 parts of patchouli, 30 parts of perilla, 50 parts of atractylodes, 150 parts of tuckahoe, 100 parts of atractylodes, 50 parts of gypsum, 30 parts of burnt hawthorn, 30 parts of burnt six Shenqu, 30 parts of burnt malt, 50 parts of earthworm, 50 parts of Cynanchum chinense, 30 parts of Cynomorium mongolicum, and 50 parts of Lemongrass, adding water for reflux extraction twice, combining the extracts, filtering, concentrating the filtrate to a relative density of 1.10-1.15, centrifuging, vacuum drying the filtrate, spray drying and pulverizing to obtain an intermediate composition, adding sucralose and dextrin, mixing, and obtaining the traditional Chinese medicine composition;

[0032] The reflux extraction includes: adding 6 times of water for the first time and extracting for 1.5 hours, and adding 4 times of water for the second time and extracting for 1.0 hour.

[0033] In some specific embodiments of the present invention, the dosage form of the drug includes: one or more of decoction, granules, capsules, tablets, oral liquid, pills, tincture, syrup, gel, spray or injection.

[0034] In some specific embodiments of the present invention, the effective dose of the drug includes 9.36-37.44 g crude drug / kg mouse body weight.

[0035] In some specific embodiments of the present invention, the administration of the drug includes one or more of oral administration, injection or external application.

[0036] The present invention provides the following beneficial effects including but not limited to:

[0037] The Chinese medicine composition of the present invention has a certain therapeutic effect for treating bacterial diarrhea, significantly improving symptoms such as fever, abdominal pain, diarrhea, vomiting, etc., and the contents of IL-6, TNF-α, IFN-γ, and IL-10 in the plasma of mice related to the Escherichia coli diarrhea test are significantly increased, indicating that the bacterial infection process causes an inflammatory response and an immune response in the body. The Chinese medicine composition particles of the present invention can regulate the levels of these cytokines, and at the same time, an inflammatory response occurs after Escherichia coli infects the intestine, causing intestinal tissue damage and a large number of neutrophils infiltrating. The Chinese medicine composition particles of the present invention can effectively reduce intestinal inflammation and protect intestinal tissue. DETAILED DESCRIPTION

[0038] The present invention discloses the use of a Chinese medicine composition in the preparation of a drug for preventing and treating bacterial infectious diarrhea. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0039] The present invention provides an application of a Chinese medicine composition in preparing a drug for treating or preventing bacterial infectious diarrhea, the Chinese medicine composition comprising: 1-100 parts of Magnolia officinalis, 1-100 parts of charred Areca nut, 1-100 parts of simmered Tsaoko fruit, 1-100 parts of Ephedra, 1-100 parts of Bitter Apricot, 1-100 parts of Notopterygium wilfordii, 1-100 parts of Ginger, 1-100 parts of Patchouli, 1-100 parts of Perilla frutescens, 1-100 parts of Atractylodes macrocephala, 1-160 parts of Poria cocos, 1-120 parts of Atractylodes macrocephala, 1-100 parts of Gypsum, 1-100 parts of charred Crataegus pinnatifida, 1-100 parts of charred Liu Shenqu, 1-100 parts of charred Malt, 1-100 parts of Pheretima, 1-100 parts of Cynanchum cyrtonema, 1-100 parts of Mianma Guanzhong, and 1-100 parts of Lepidium tiliaceum. The application may be any method that is beneficial to improving the corresponding symptoms of the patient, including treatment or prevention.

[0040] Furthermore, the Chinese medicine composition includes: 10-80 parts of Magnolia officinalis, 10-80 parts of scorched Areca catechu, 10-80 parts of stewed Tsaoko fruit, 10-60 parts of Ephedra sinica, 10-60 parts of Bitter Apricot, 10-80 parts of Notopterygium wilfordii, 10-60 parts of Ginger, 10-80 parts of Patchouli, 10-60 parts of Cyperus rotundus, 10-80 parts of Atractylodes macrocephala, 10-160 parts of Poria cocos, 10-120 parts of Atractylodes macrocephala, 10-80 parts of Gypsum, 10-50 parts of scorched Crataegus pinnatifida, 10-80 parts of scorched Liu Shenqu, 10-60 parts of scorched malt, 10-80 parts of Pheretima, 10-80 parts of Cynanchum cyrtonema, 10-60 parts of Lemongrass seeds.

[0041] Preferably, the Chinese medicine composition comprises: 30-50 parts of Magnolia officinalis, 20-30 parts of scorched Areca nut, 30-50 parts of stewed Tsaoko fruit, 20-30 parts of Ephedra, 20-30 parts of Bitter Apricot, 30-50 parts of Notopterygium wilfordii, 30-50 parts of Ginger, 30-50 parts of Patchouli, 20-30 parts of Cyperus rotundus, 30-50 parts of Atractylodes macrocephala, 120-150 parts of Poria, 80-100 parts of Atractylodes macrocephala, 30-50 parts of Gypsum, 20-30 parts of scorched Crataegus pinnatifida, 30-50 parts of scorched Liu Shenqu, 20-30 parts of scorched malt, 30-50 parts of Pheretima, 30-50 parts of Cynanchum cyrtonema, and 30-50 parts of Lemongrass seeds.

[0042] Preferably, the Chinese medicine composition includes: 50 parts of Magnolia officinalis, 30 parts of scorched areca nut, 30 parts of stewed tsaoko fruit, 20 parts of ephedra, 30 parts of bitter almond, 50 parts of Notopterygium wilfordii, 50 parts of ginger, 50 parts of patchouli, 30 parts of Cyperus rotundus, 50 parts of Atractylodes macrocephala, 150 parts of Poria cocos, 100 parts of Atractylodes macrocephala, 50 parts of gypsum, 30 parts of scorched hawthorn, 30 parts of scorched Liu Shenqu, 30 parts of scorched malt, 50 parts of earthworm, 50 parts of Cynomorium wilfordii, 30 parts of Cynomorium fortunei, and 50 parts of Lemongrass seeds.

[0043] The above-mentioned Chinese medicine composition can be directly ground into powder by raw materials, or can be an extract or other forms prepared by conventional means.

[0044] Specifically, the dosage forms of the aforementioned bacterial infectious diarrhea drugs include oral dosage forms, injection dosage forms or external dosage forms.

[0045] The drug of the present invention is prepared by weighing the raw material drug in proportion, adding pharmaceutically acceptable excipients such as fillers, disintegrants, lubricants, suspending agents, adhesives, sweeteners, flavoring agents, preservatives, bases, etc., and preparing according to conventional production methods in the pharmaceutical field to prepare conventional dosage forms acceptable to pharmacy, including but not limited to decoctions, granules, capsules, tablets, oral liquids, pills, tinctures, syrups, suppositories, gels, sprays, and injections.

[0046] The present invention also provides a method for preparing any of the aforementioned Chinese medicine compositions, the method comprising:

[0047] Weigh Magnolia officinalis, scorched areca nut, stewed tsaoko, ephedra, bitter almond, notopterygium wilfordii, ginger, patchouli, perilla, atractylodes, Poria, atractylodes macrocephala, gypsum, scorched hawthorn, scorched Liushenqu, scorched malt, earthworm, Cynanchum chinense, Cynomorium morganianum, and Semen tiliaceum, extract twice with water, add 6 times the amount of water for the first time and extract for 1.5 h, add 4 times the amount of water for the second time and extract for 1.0 h, combine the extracts, filter, concentrate the filtrate to a relative density of 1.10-1.15, centrifuge and filter, and dry the filtrate.

[0048] Specifically, the above preparation method has the following steps:

[0049] Take 50 parts of Magnolia officinalis, 30 parts of charred Areca nut, 30 parts of stewed Tsaoko fruit, 20 parts of Ephedra, 30 parts of Bitter Apricot, 50 parts of Notopterygium, 50 parts of Ginger, 50 parts of Patchouli, 30 parts of Perilla frutescens, 50 parts of Atractylodes, 150 parts of Poria, 100 parts of Atractylodes, 50 parts of Gypsum, 30 parts of charred Crataegus, 30 parts of charred Liu Shenqu, 30 parts of charred Malt, 50 parts of Pheretima, 50 parts of Cynanchum indica, 30 parts of Cynomorium mongolicum, and 50 parts of Semen Lepidii, add water and reflux extract twice, add 6 times of water for the first time, extract for 1.5 h, add 4 times of water for the second time, extract for 1.0 h, combine the extracts, filter, concentrate the filtrate to a relative density of 1.10-1.15, centrifuge and filter, vacuum dry the filtrate, spray dry and crush to obtain an intermediate composition, add sucralose and dextrin, mix well, and obtain granules.

[0050] The composition of the present invention has a certain therapeutic effect on treating bacterial diarrhea, significantly improving symptoms such as fever, abdominal pain, diarrhea, and vomiting. The levels of IL-6, TNF-α, IFN-γ, and IL-10 in the plasma of mice related to the Escherichia coli diarrhea test are significantly increased, indicating that the bacterial infection process causes inflammatory and immune responses in the body. The particles of the composition of the present invention can regulate the levels of these cytokines. At the same time, inflammatory reactions occur after Escherichia coli infects the intestines, causing intestinal tissue damage and a large number of neutrophils infiltrating. The particles of the composition of the present invention can effectively reduce intestinal inflammation and protect intestinal tissue.

[0051] The following experiments, if no specific conditions are specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The APIs or excipients, reagents or instruments used, if the manufacturer is not specified, are all conventional products that can be purchased commercially. Unless otherwise specified, all percentages, ratios, proportions or parts are by weight.

[0052] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the present invention.

[0053] Unless otherwise specified, the raw materials and reagents used in the application of the Chinese medicine composition provided by the present invention in the preparation of drugs for preventing and treating bacterial infectious diarrhea can all be purchased from the market.

[0054] The present invention will be further described below in conjunction with embodiments:

[0055] Preparation of Chinese medicine composition granules

[0056] The Chinese medicine composition of this embodiment is prepared from the following raw materials: 50 grams of Magnolia officinalis, 30 grams of scorched betel nut, 30 grams of stewed tsaoko, 20 grams of ephedra, 30 grams of bitter almond, 50 grams of Notopterygium wilfordii, 50 grams of ginger, 50 grams of patchouli, 30 grams of Cyperus rotundus, 50 grams of Atractylodes macrocephala, 150 grams of Poria, 100 grams of Atractylodes macrocephala, 50 grams of gypsum, 30 grams of scorched hawthorn, 30 grams of scorched Liu Shenqu, 30 grams of scorched malt, 50 grams of earthworm, 50 grams of Cynomorium fortunei, 30 grams of Cynomorium cyrtonema, and 50 grams of Lemongrass.

[0057] The Chinese medicine composition is prepared according to the following method:

[0058] Take the above medicinal materials, add water and reflux extract twice, add 6 times water for the first time, extract for 1.5 hours, add 4 times water for the second time, extract for 1.0 hours, combine the extracts, filter, concentrate the filtrate to a relative density of 1.10-1.15, centrifuge and filter, vacuum dry the filtrate, spray dry and crush to obtain an intermediate composition, add sucralose and dextrin, mix well, and obtain the Chinese medicine composition granules.

[0059] Effect of Examples on Escherichia coli Infectious Diarrhea

[0060] Experimental Materials

[0061] Animals: BABL / C mice, SPF grade, male, 6-8 weeks old, weighing 18-20 g, a total of 60 mice were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0062] Drug: Enrofloxacin (Batch No.: 93106-60-6) Sichuan Weikeqi Biotechnology Co., Ltd.

[0063] Test drug: the Chinese medicine composition granules prepared in the preparation example, provided by Jiangsu Kangyuan Pharmaceutical Co., Ltd. Drug dosage: The daily dosage of crude drugs for humans is 288 g, and the clinical dosage for adults (calculated as 70 kg) is 4.1 g crude drugs / kg. The equivalent dosage for mice is calculated to be 37.44 g crude drugs / kg. This dosage is set as the high dosage, and the low, medium, and high dosage groups are set according to the dosage ratio of 0.25:0.5:1, with dosages of 9.36, 18.72, and 37.44 g crude drugs / kg, respectively, which are equivalent to 2.3, 4.6, and 9.1 times the clinical dosage, respectively.

[0064] Reagents: Enteropathogenic Escherichia coli EPEC, No.: BNCC216733, purchased from China Industrial Microbiological Culture Collection Management Center, Bacteriological peptone (Batch No.: LP0037B) from Guangzhou Hongzhou Experimental Equipment Technology Co., Ltd., Technical agar powder (Batch No.: 028990) from Jiangsu Kangcheng Baiao Biotechnology Co., Ltd., Beef extract (Batch No.: 050060A) from Guangdong Huankai Microbiological Technology Co., Ltd., MPO ELISA kit (Batch No.: 69-21015) from Wuhan Merck Biotechnology Co., Ltd., TNF-α ELISA kit (lot number: EIA-0082) Shanghai Aleisa Biotechnology Co., Ltd., IL-1β (lot number: EIA-0058) Shanghai Aleisa Biotechnology Co., Ltd., IL-6 (EIA-0111) Shanghai Aleisa Biotechnology Co., Ltd., IFN-γ (EIA-0133) Shanghai Aleisa Biotechnology Co., Ltd., IL-10 (EIA-0126) Shanghai Aleisa Biotechnology Co., Ltd., paraformaldehyde (V900894-100G) Merck.

[0065] Instruments: biochemical incubator (LRH-15B, Shanghai Boxun Industrial Co., Ltd.), SW-CJ-2FD clean bench (Suzhou Antai Air Technology Co., Ltd.), biological microscope (ECLIPSEE200, Nikon, Japan), MULITSKAN Sky microplate reader (Thermo Fisher Scientific, USA), MINIP-2500 microplate centrifuge, MINI-4K microcentrifuge (Hangzhou Miou Instrument Co., Ltd.), 5418R low-temperature high-speed centrifuge (Eppendorf, Germany), Nano-100 micro-spectrophotometer (Hangzhou Aosheng Instrument Co., Ltd.).

[0066] Experimental methods

[0067] Preparation of culture medium

[0068] 5 g sodium chloride, 3 g beef extract, 10 g peptone, dilute to 1000 mL, shake well, adjust pH to 7.4 with NaOH solution (1 mol / L); store in a 4°C refrigerator for later use. Add 13 g technical agar powder to the mixed liquid, autoclave at 121°C for 20 min, seal after cooling, culture in a 37°C constant temperature incubator for 18-24 h, and store at 4°C to obtain a sterile culture medium.

[0069] Preparation of E. coli suspension

[0070] The freeze-dried bacterial powder was dissolved in 0.5 mL of sterile culture medium, and the dissolved bacterial suspension was transferred to 4-5 mL of liquid culture medium and mixed evenly. It was inoculated in nutrient agar medium and placed in a 37°C constant temperature incubator for activation. The activated Escherichia coli colonies were selected and inoculated in 20 mL of nutrient liquid culture medium, and cultured at 37°C for 18-24 h. The bacteria were counted by pouring plate method, and the number of colonies between 30 and 300 on the plate was selected. The number of colonies per mL of bacterial solution (1.0×10 9 CFU / mL).

[0071] Establishment of Escherichia coli diarrhea model

[0072] 60 SPF male BABL / C mice aged 6 to 8 weeks were randomly divided into six groups, namely, blank control group, model group, high, medium and low dose groups of the Chinese medicine composition granules of the present invention (9.36 g crude drug / kg, 18.72 g crude drug / kg, 37.44 g crude drug / kg), and enrofloxacin group (5 mg / kg), with 10 mice in each group. Except for the blank control group, each mouse was intraperitoneally injected with about 10 mL / kg of Escherichia coli, and the blank control group was given an equal volume of normal saline. After the onset of diarrhea symptoms, the status of mice in each group, such as mental state, fecal morphology, respiration, diet, etc., was observed. After the onset of diarrhea symptoms, mice in the high, medium and low dose groups were gavaged with different doses of the Chinese medicine composition granules of the present invention, the normal group and the model group were gavaged with equal volumes of normal saline, and the enrofloxacin group was gavaged with enrofloxacin solution according to the dose, and the drug was administered once a day.

[0073] Mouse detection indicators: ① Blood was collected from mice injected with Escherichia coli on the 9th day, centrifuged at 3000 r / min for 3 min, and the serum was separated and stored in a -20℃ refrigerator. According to the instructions of the Elisa kit, the levels of serum inflammatory factors TNF-α, IFN-γ, IL-6, and IL-10 were determined. ② Two sections of duodenum were collected from mice in each group, one section was fixed in paraformaldehyde, and the other end of the duodenum tissue was frozen. Cold physiological saline was used as the homogenization medium to prepare intestinal homogenate at a ratio of 1:9, and MPO activity was detected according to the instructions of the MPO kit.

[0074] The results showed that compared with the blank control group, the IL-6 content in the model group was extremely significantly increased (P<0.01); compared with the model group, the IL-6 content in the high-, medium-, low-dose groups and enrofloxacin groups was extremely significantly decreased (P<0.01), and the IL-6 levels in the high-dose group, enrofloxacin, medium-dose group and low-dose group increased in turn; compared with the control group, the TNF-α level of mice in the model group was extremely significantly increased (P<0.01); the TNF-α level of mice in each treatment group showed different degrees of recovery, among which the TNF-α level of mice in the high-dose group recovered significantly and was significantly lower than that in the model group (P<0.05). Compared with the control group, the levels of IFN-γ (P<0.01) and IL-10 (P<0.01) in the model group were extremely significantly increased; compared with the model group, the levels of IFN-γ and IL-10 in mice in each treatment group decreased to varying degrees, among which the IFN-γ level in mice in the high-dose group decreased significantly (P<0.05), the IFN-γ level in mice in the medium and low-dose groups decreased but the difference was not significant compared with the model group (P>0.05), the IL-10 content in mice in the high-dose group decreased significantly (P<0.05), and the difference between the medium and low-dose groups and the model group was not significant (P>0.05). See Table 1.

[0075] After intestinal infection with E. coli, the MPO activity in the intestinal tissue of mice in the model group was significantly increased compared with that in the control group (P<0.01); compared with the model group, the MPO activity levels of the intestinal groups decreased to varying degrees after treatment with the granules of the composition of the present invention, among which the MPO activity of the high-dose group, enrofloxacin group (P<0.01) and medium-dose group (P<0.05) was significantly reduced, and the MPO activity of mice in the low-dose group decreased, but the difference was not significant compared with the model group. See Table 2.

[0076] Table 1 Effects on the levels of serum inflammatory factors in diarrheal mice

[0077]

[0078] Note: Compared with the blank control group, * P<0.05, ** P<0.01. Compared with the model group ＃ P<0.05, ＃＃ P<0.01.

[0079] Table 2 Effects on MPO levels in mice with diarrhea

[0080]

[0081] Note: Compared with the blank control group, * P<0.05, ** P<0.01. Compared with the model group ＃ P<0.05, ＃＃P<0.01.

[0082] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Use of a Chinese medicine composition in the preparation of a drug for treating and / or preventing bacterial infectious diarrhea; The Chinese medicine composition comprises: The invention discloses one or a combination of two or more of 1-100 parts of Magnolia officinalis bark, 1-100 parts of burnt Areca catechu, 1-100 parts of stewed Tsaoko fruit, 1-100 parts of Ephedra sinica, 1-100 parts of Bitter Apricot, 1-100 parts of Notopterygium wilfordii, 1-100 parts of Ginger, 1-100 parts of Patchouli, 1-100 parts of Perilla frutescens, 1-100 parts of Atractylodes macrocephala, 1-160 parts of Poria cocos, 1-120 parts of Atractylodes macrocephala, 1-100 parts of Gypsum, 1-100 parts of burnt Crataegus pinnatifida, 1-100 parts of burnt Liu Shenqu, 1-100 parts of burnt malt, 1-100 parts of Pheretima, 1-100 parts of Cynanchum cyrtonema, 1-100 parts of Cynomorium wilfordii and / or 1-100 parts of Semen Lepidium.

2. The use according to claim 1, characterized in that The treatment and / or prevention of bacterial infectious diarrhea includes any of the following: (I) Improve fever; and / or (II), improve abdominal pain; and / or (III) Improve diarrhea; and / or (IV), improve vomiting; and / or (V) Reduce intestinal inflammation.

3. The use according to claim 2, characterized in that The reducing of intestinal inflammation includes regulating cytokine levels and / or reducing inflammatory cell infiltration.

4. The use according to claim 3, characterized in that The regulating cytokine level comprises any of the following: (I), reducing the level of IL-6; and / or (II), reducing the content of TNF-α; and / or (III), reducing the content of IFN-γ; and / or (IV) Reduce the content of IL-10.

5. The use according to claim 3 or 4, characterized in that: The reduction of inflammatory cell infiltration includes reducing the MPO activity level in intestinal tissue.

6. The use according to any one of claims 1 to 5, characterized in that The Chinese medicine composition comprises: 10-80 parts of Magnolia officinalis, 10-80 parts of charred Areca catechu, 10-80 parts of stewed Tsaoko fruit, 10-60 parts of Ephedra sinica, 10-60 parts of Apricot kernel, 10-80 parts of Notopterygium wilfordii, 10-60 parts of ginger, 10-80 parts of Patchouli, 10-60 parts of Cyperus rotundus, 10-80 parts of Atractylodes macrocephala, 10-160 parts of Poria cocos, 10-120 parts of Atractylodes macrocephala, 10-80 parts of Gypsum, 10-50 parts of charred Crataegus pinnatifida, 10-80 parts of charred Liu Shenqu, 10-60 parts of charred malt, 10-80 parts of Pheretima, 10-80 parts of Cynomorium wilfordii, 10-60 parts of Cynomorium fortunei and / or 10-80 parts of Lepidium chinense.

7. The use according to any one of claims 1 to 6, characterized in that The Chinese medicine composition comprises: 30-50 parts of Magnolia officinalis, 20-30 parts of burnt betel nut, 30-50 parts of stewed tsaoko, 20-30 parts of ephedra, 20-30 parts of bitter almond, 30-50 parts of notopterygium, 30-50 parts of ginger, 30-50 parts of patchouli, 20-30 parts of perilla, 30-50 parts of atractylodes, 120-150 parts of tuckahoe, 80-100 parts of atractylodes, 30-50 parts of gypsum, 20-30 parts of burnt hawthorn, 30-50 parts of burnt six-spirit song, 20-30 parts of burnt malt, 30-50 parts of earthworm, 30-50 parts of cynanchum chinense, 20-30 parts of cynanchum chinense and / or 30-50 parts of scutellaria baicalensis. Preferably, one or a combination of two or more of 50 parts of Magnolia officinalis, 30 parts of scorched Areca nut, 30 parts of stewed Tsaoko fruit, 20 parts of Ephedra, 30 parts of Bitter Apricot, 50 parts of Notopterygium wilfordii, 50 parts of Ginger, 50 parts of Patchouli, 30 parts of Perilla frutescens, 50 parts of Atractylodes lancea, 150 parts of Poria, 100 parts of Atractylodes macrocephala, 50 parts of Gypsum, 30 parts of scorched Crataegus pinnatifida, 30 parts of scorched Liu Shenqu, 30 parts of scorched malt, 50 parts of Pheretima, 50 parts of Cynanchum chinense, 30 parts of Cynomorium mongolicum and / or 50 parts of Semen Lepidium.

8. The use according to any one of claims 1 to 7, characterized in that The preparation method of the traditional Chinese medicine composition comprises: taking one or a combination of two or more of magnolia bark, burnt betel nut, stewed tsaoko, ephedra, bitter almond, notopterygium wilfordii, ginger, patchouli, perilla, atractylodes, tuckahoe, atractylodes macrocephala, gypsum, burnt hawthorn, burnt six-shenqu, burnt malt, earthworm, cynanchum chinense, mianma guanzhong and / or scutellaria baicalensis seed in a prescribed amount, refluxing extraction, filtering, concentrating, centrifuging, drying and pulverizing to obtain an intermediate composition; Taking the intermediate composition and mixing it with pharmaceutically acceptable excipients to prepare the Chinese medicine composition; The reflux extraction includes: adding 6 times of water for the first time and extracting for 1.5 hours, and adding 4 times of water for the second time and extracting for 1.0 hour.

9. The use according to any one of claims 1 to 8, characterized in that The effective dose of the drug includes 9.36-37.44 g crude drug / kg mouse body weight.

10. The use according to any one of claims 1 to 9, characterized in that The administration method of the drug includes: one or more of oral administration, injection or external application.

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