Application of sophocarpidine in preparation of intestinal microbiota regulator and anti-escherichia coli medicine

Abstract

The invention relates to the field of biological pharmacy, in particular to application of matrine in preparation of an intestinal microbiota regulator and an anti-escherichia coli medicine. The invention provides application of matrine in preparation of an intestinal microbiota regulator and an anti-Escherichia coli medicine, and matrine has the following advantages in application: (1) an active compound of a plant source has the characteristics of specificity and naturalness, and is friendly to the ecological environment; (2) the chemical structure and chemical characteristics are known, and the compound plays a role together with the immune system and the like of a host; (3) the main mode of acting on pathogens is a weakening and inhibiting mechanism or a synergistic effect with other substances, so that the drug resistance of pathogenic microorganisms is not easily caused, and the effectiveness of long-term use is ensured; and (4) pathogenic bacteria cannot be directly and completely killed generally, and corresponding pathogenic microorganisms are inhibited in a way of controlling an ecological steady state, so that the survival, replication and proliferation of the microbial community cannot be seriously influenced.

Description

Technical Field

[0001] The invention relates to the field of biopharmaceuticals, and in particular to the application of matrine in the preparation of an intestinal microbiome regulator and an anti-Escherichia coli drug. Background Art

[0002] Microorganisms such as bacteria, viruses and parasites in the environment mostly exist in a conditionally pathogenic way. Once the body's resistance decreases, these pathogenic microorganisms will take advantage of the situation and induce infection and outbreak of diseases. Humans and animals are always facing the threat of diseases during their growth and development. Among the multiple factors that cause diseases, pathogenic microorganisms often dominate. Among the many types of pathogens, Escherichia coli is a very common and widespread species, which has been explored and studied from different angles, such as phylogenetic analysis and O serotype diversity distribution. Due to its universal existence in the body, pan-host and strong tolerance to physical and chemical factors, Escherichia coli is easier to survive, replicate and reproduce in the environment, which not only causes great harm to human and animal health, but also causes huge economic losses to animal production, seriously endangers food and public health safety, and hinders the development of animal husbandry.

[0003] Antibiotics are a conventional means of treating bacterial diseases, but the widespread, large-scale and irrational use of antibiotics has led to the emergence of drug-resistant bacteria and destroyed the normal balance of the body's flora. At present, the problem of bacterial resistance in clinical practice has become quite serious, and the effect of traditional conventional antibiotics in treating bacterial infections is not ideal. In view of the practical difficulties of the numerous serotypes of Escherichia coli and the poor cross-specificity of species, it is impossible to play a comprehensive and effective preventive role through vaccine immunization. As one of the important immune organs of the body, the intestinal microbiota can play a role in resisting and alleviating bacterial infections by regulating the composition and proportion of the intestinal flora. In the study of intestinal flora, oral intake of food, drugs and exogenous substances is considered to be an important reason for the significant differences in the abundance and composition of intestinal flora. At the same time, a large number of studies have found that the intestinal flora is significantly associated with host immunity, nerve center, nutritional metabolism, endocrine, tumors, anti-inflammation and anti-infection. However, further research is needed to study the intrinsic mechanism of this regulation and correlation. In the external environment and host intestinal environment, it is known that bacteria are affected by factors such as temperature, humidity, Ph, nutrients, and drugs. Whether these are related to the host intestinal microbiota and the extent of regulatory influence are hot topics of research.

[0004] Active natural compounds in traditional Chinese medicine have multiple advantages such as wide sources, diverse activities, simple preparation and low prices. They can effectively make up for the disadvantages of poor efficacy and delayed research and development of existing antibiotics, and have research and development advantages and future prospects. Natural active ingredients from traditional Chinese medicine regulate the structure of the host's intestinal microbial community, participate in the host's physiological, biochemical and immune response processes, and have a profound impact on maintaining the stability of the host ecosystem. In this field, the key to the problem is to discover active natural compounds that regulate the host's intestinal flora. If the substances that regulate the host's microbial community and resist bacterial infection can be fully explored, their action characteristics can be demonstrated and identified by regulating the abundance of the host's intestine, improving the host's ability to clear pathogens and the mechanism of action, and then deeply explored and comprehensively utilized, it is crucial to regulate the host's intestinal health and prevent and control bacterial infections. Therefore, the regulation of intestinal microbiota and the discovery and application of related drugs have very important practical significance and innovative value. Summary of the invention

[0005] The purpose of the present invention is to provide the use of matrine in the preparation of an intestinal microbial flora regulator and an anti-Escherichia coli drug to solve the problems of the above-mentioned prior art. The present invention first discovered that matrine has an anti-Escherichia coli effect and can also regulate the intestinal microbial flora of mice infected with Escherichia coli, providing a new idea for the preparation of drugs for anti-Escherichia coli and regulating intestinal microbial flora.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The present invention provides application of matrine in preparing an intestinal microbiota regulator.

[0008] The matrine of the present invention, also known as matricine, is a white solid powder that is soluble in water, benzene, chloroform, methanol, ethanol and slightly soluble in solvents such as petroleum ether. The CAS registration number of the matrine is 519-02-8, and the molecular formula is C 15 H 24 N 2 O, molecular weight is 248.364, chemical structure is shown in formula I:

[0009]

[0010] Preferably, the intestinal microbiota regulator achieves the effect of regulating the intestinal microbiota by increasing the abundance of Lactobacillus murine.

[0011] The invention provides an intestinal microbiota regulator, which comprises matrine and pharmaceutically acceptable auxiliary materials.

[0012] Preferably, the intestinal microbiota regulator achieves the effect of regulating the intestinal microbiota by increasing the abundance of Lactobacillus murine.

[0013] The invention provides application of matrine in preparing an intestinal lactobacillus murine regulator.

[0014] The invention provides an intestinal lactobacillus rat regulator, which comprises matrine and pharmaceutically acceptable auxiliary materials.

[0015] The invention provides application of matrine in preparing medicine for resisting Escherichia coli.

[0016] Preferably, the anti-E. coli drug achieves its anti-E. coli effect by reducing the number of E. coli colonies colonizing the liver and regulating intestinal microbial flora.

[0017] The invention provides an anti-Escherichia coli drug, which comprises matrine and pharmaceutically acceptable auxiliary materials.

[0018] Preferably, the anti-E. coli drug achieves its anti-E. coli effect by reducing the number of E. coli colonies colonizing the liver and regulating intestinal microbial flora.

[0019] The present invention discloses the following technical effects:

[0020] Matrine is an active natural compound with the effects of regulating intestinal microbial communities and protecting against infection. It is derived from the dried roots, plants and fruits of the leguminous plant Sophora flavescens, and is an alkaloid substance extracted by organic solvents such as ethanol. The present invention selects enterotoxigenic Escherichia coli (ETEC) as a representative of pathogenic strains, and studies the effect of matrine in inhibiting Escherichia coli infection by constructing a mouse ETEC infection model. Studies have shown that matrine effectively alleviates the weight loss trend of ETEC-infected mice and improves the weight gain level of mice; bacterial 16S rRNA gene sequencing results show that matrine effectively regulates the composition and abundance of intestinal microbiota in infected mice and promotes the replication and proliferation of beneficial bacteria; matrine significantly reduces the number of colonies colonized in the liver of ETEC-infected mice; and effectively improves the host's ability to clear pathogens.

[0021] At the same time, it also has the following advantages in fighting E. coli and regulating intestinal microbial flora:

[0022] (1) Active compounds derived from plants are specific, natural, and friendly to the ecological environment; (2) The chemical structure and chemical properties are known, and they work together with the host's immune system; (3) The main way they act on pathogens is through weakening and inhibition mechanisms, or synergistic effects with other substances, making it less likely for pathogens to develop drug resistance, thereby ensuring their effectiveness during long-term use; (4) They generally do not directly and completely kill pathogens, but inhibit the corresponding pathogens by controlling ecological homeostasis, which will not seriously affect the survival, replication, and proliferation of the microbial community.

[0023] In summary, matrine is significantly different from general chemical drugs that are highly toxic, highly residual, and easily induced to induce drug resistance. It is a very green and environmentally friendly natural compound, and it can effectively inhibit E. coli and regulate the intestinal microbial flora of mice infected with E. coli. Therefore, for the future, the present invention has very important guiding significance in both the extraction and preparation of antibacterial infection drugs and the chemical synthesis of antibacterial infection drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0025] Figure 1 The weight loss levels of mice infected with ETEC under different treatments;

[0026] Figure 2 The colonies were colonized in the livers of mice infected with ETEC under different treatments; the control group was the infection control group;

[0027] Figure 3 Abundance and composition of intestinal microbiota in mice infected with ETEC under different treatments. DETAILED DESCRIPTION

[0028] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0029] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0030] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0031] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0032] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0033] Example 1

[0034] 1. Establishment of an enterotoxigenic Escherichia coli (ETEC) infection mouse model and challenge and treatment

[0035] (1) Pick a single colony of enterotoxigenic E. coli ETEC-1 (manufacturer: TSTO, catalog number: TS326593) and place it in 2 mL of fresh LB liquid medium, and culture it overnight at 37°C and 200 rpm in a shaker for 13 h. The next day, expand the culture to 20 mL of fresh LB liquid medium at a ratio of 1:100, and culture it in a shaker at 37°C and 200 rpm for 4 h. 600nm When the absorbance at a wavelength of 600 nm is about 0.6, the OD of the bacterial solution is 600nm Adjust to 0.1 as bacterial suspension for later use.

[0036] (2) 30 6-8 week old SPF female Balb / c mice (weight 16-18 g) were selected, adaptively fed for 3 days, and fasted for 1 hour before infection. The mice that had completed the above treatment were randomly divided into three groups, namely, a blank control group (no treatment, isolated and fed alone), an infection control group (no drug treatment after infection), and a matrine treatment group (matrine treatment after infection, the dosage was 20 mg / kg), with 10 mice in each group. The bacterial suspension prepared above was taken and attacked SPF female Balb / c mice by oral gavage. The dosage of enterotoxigenic E. coli ETEC-1 was 1×10 9 CFUs / mouse, and the mouse infection model of enterotoxigenic Escherichia coli was established.

[0037] In this study, enterotoxigenic Escherichia coli was selected as the challenge strain, and SPF mice were challenged by oral gavage. Matrine was used as a candidate therapeutic drug, and the infected mice were treated with matrine orally. The weight of mice in each treatment group was weighed and counted, and the number of liver colonies was detected on the 5th day after infection to evaluate the protective effect of matrine on infected mice at the in vivo level; at the same time, the intestinal microbial flora of infected mice was investigated.

[0038] 2. Results

[0039] (1) Effect of matrine on body weight of mice infected with enterotoxigenic Escherichia coli

[0040] The mouse model of enterotoxigenic Escherichia coli infection was established as described above. The matrine-treated group was orally administered once a day for 5 consecutive days. The mice were weighed every day for 5 consecutive days after infection, and the average weight of mice in each group was calculated and statistically analyzed. The statistical results are shown in the figure. Figure 1 Compared with the infected control group, matrine treatment significantly alleviated the weight loss trend of infected mice, and the average weight gain of mice in the matrine treatment group was not significantly different from that in the blank control group.

[0041] (2) Effect of matrine on liver colonization of mice infected with enterotoxigenic Escherichia coli

[0042] On the 5th day after infection, liver tissues of mice in each group were collected, weighed and ground to prepare tissue homogenate, diluted with sterile PBS in multiple ratios and then spread on LB solid agar culture plates. After incubation at 37°C for 12 hours, colony counts were performed. The colony count results were as follows: Figure 2 The results showed that compared with the infected control group, the number of liver colonies in the matrine-treated mice was significantly reduced.

[0043] (3) Effects of matrine on intestinal microbiota of mice infected with enterotoxigenic Escherichia coli

[0044] On the 5th day after infection, 1g of feces was collected from each group of mice. 16SrRNA gene sequencing was performed by extracting bacterial DNA from the feces of each group of mice. Combining sequencing method data with bioinformatics methods, the composition and abundance changes of the intestinal microbiota of each group of mice were analyzed. The results are as follows: Figure 3 The results showed that on the 5th day after infection, the composition and abundance of the intestinal microbial community in the mice treated with matrine changed significantly compared with the infected control group, especially the abundance of probiotics such as Lactobacillus murinus increased significantly. It can be seen that matrine can regulate the abundance and composition of the intestinal microbial flora in mice infected with ETEC.

[0045] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of matrine in the preparation of intestinal microbiota regulators.

2. The use according to claim 1, characterized in that: The intestinal microbiota regulator achieves the effect of regulating the intestinal microbiota by increasing the abundance of Lactobacillus murine.

3. An intestinal microbiome regulator, characterized in that The intestinal microbial flora regulator comprises matrine and pharmaceutically acceptable auxiliary materials.

4. The intestinal microbiota regulator according to claim 3, characterized in that The intestinal microbiota regulator achieves the effect of regulating the intestinal microbiota by increasing the abundance of Lactobacillus murine.

5. Application of matrine in the preparation of intestinal Lactobacillus murine regulator.

6. An intestinal Lactobacillus ratus regulator, characterized in that The intestinal lactobacillus rat regulator comprises matrine and pharmaceutically acceptable auxiliary materials.

7. The application of matrine in the preparation of anti-Escherichia coli drugs.

8. The use according to claim 7, characterized in that: The anti-E. coli drug achieves the anti-E. coli effect by reducing the colonization number of E. coli in the liver and regulating the intestinal microbial flora.

9. An anti-Escherichia coli drug, characterized in that: The medicine comprises matrine and pharmaceutically acceptable auxiliary materials.

10. The drug according to claim 9, characterized in that The anti-E. coli drug achieves the anti-E. coli effect by reducing the colonization number of E. coli in the liver and regulating the intestinal microbial flora.

Images