CN110123806B - Application of epigallocatechin gallate in preparation of anti-streptococcus suis drugs - Google Patents
Application of epigallocatechin gallate in preparation of anti-streptococcus suis drugs Download PDFInfo
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- CN110123806B CN110123806B CN201910474855.0A CN201910474855A CN110123806B CN 110123806 B CN110123806 B CN 110123806B CN 201910474855 A CN201910474855 A CN 201910474855A CN 110123806 B CN110123806 B CN 110123806B
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
- A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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Abstract
The invention belongs to the technical field of veterinary drugs, and particularly relates to application of epigallocatechin gallate in preparation of a streptococcus suis resistant drug. The applicant finds that the epigallocatechin gallate has the effects of inhibiting the growth of the streptococcus suis and killing the streptococcus suis, the minimum inhibitory concentration of the epigallocatechin gallate to the streptococcus suis is 512 mu g/ml, and the minimum bactericidal concentration of the epigallocatechin gallate to the streptococcus suis is 1024 mu g/ml, so that the toxic and side effects and the potential safety hazards of vaccines, chemical drugs and antibiotics are overcome, and the epigallocatechin gallate has wide application prospects and development values in the fields of food processing, medicine and breeding industry.
Description
Technical Field
The invention belongs to the technical field of veterinary drugs, and particularly relates to application of epigallocatechin gallate in preparation of a streptococcus suis resistant drug.
Background
Streptococcus suis is a zoonotic infectious disease pathogen and can cause meningitis, septicemia, arthritis, endocarditis and pneumonia, resulting in the death of pigs and even humans due to infection. The swine streptococcosis not only causes great economic loss to the pig industry, but also poses great threat to the public health safety of human beings. Streptococcus suis is divided into 33 serotypes according to the specificity of bacterial capsular polysaccharide antigens, and among the serotypes, streptococcus suis type 2 is the most pathogenic and the dominant serotype which is often isolated from pigs who die of illness.
The control of swine streptococcosis is mainly carried out by prevention and treatment with antibiotics. However, in recent years, with the large, unjustified use of antibiotics, resistance to bacterial pathogens has become very severe. Clinical experimental data show that streptococcus suis isolated in different countries and regions has different degrees of drug resistance. In the streptococcus suis isolated by Marie, a french scholarer, almost all strains are sensitive to penicillin G, amoxicillin, ceftiofur, florfenicol, gentamicin, bacitracin zinc, and the most serious drug resistance is doxycycline, macrolides and lincosamides. The Korean scholars Dong tested that 55 strains of Streptococcus suis isolated from domestic pigs were resistant to tetracycline, gentamicin and spectinomycin, which were commonly used in clinical practice. 53 streptococcus suis separated from Shandong diseased pigs by Chinese scholars have high drug resistance to common antibiotics and complex drug resistance conditions, particularly the drug resistance to tetracycline, streptomycin, kanamycin and lincomycin is over 80 percent, the drug resistance of sulfonamides reaches 73 percent, and the drug resistance of beta-lactam drugs also reaches 20 percent. The 32 streptococcus suis separated in Guangxi has very common drug resistance and wide drug resistance spectrum, has drug resistance to amoxicillin, has the drug resistance rate of 100 percent, and has drug resistance to more than 3 antibiotics and multiple drug resistance. Therefore, the search for new antibiotic substitutes and the development of new antibacterial drugs have been urgently needed. The tea polyphenol is a polyphenol compound separated and purified from tea leaves, and has excellent antibacterial activity on nearly one hundred pathogenic bacteria of more than ten species groups in nature.
Disclosure of Invention
The invention aims to solve the technical problem of overcoming the serious drug resistance of the existing streptococcus suis and provides application of epigallocatechin gallate (EGCG) in preparing a drug for resisting streptococcus suis. The substance has good bacteriostatic and bactericidal effects on the streptococcus suis and has good application prospect in the aspect of preventing and treating the streptococcus suis.
In order to achieve the purpose, the following technical scheme is adopted:
application of epigallocatechin gallate (EGCG) in preparing medicines for resisting Streptococcus suis includes application in preparing medicines for treating or preventing Streptococcus suis infection, including inhibiting growth of in vitro Streptococcus suis by using the substance, or directly killing Streptococcus suis; or can be prepared into a preparation for the medicines for the diseases infected by the streptococcus suis.
In the above application, preferably, the streptococcus suis is streptococcus suis type 2.
Compared with the prior art, the invention has the following advantages and effects:
the invention discovers that the epigallocatechin gallate can be used for resisting streptococcus suis for the first time. The invention proves that the epigallocatechin gallate has good antibacterial effect on the streptococcus suis through a plurality of methods, lays a theoretical foundation for developing novel medicines for resisting the streptococcus suis, even provides candidate substances for the discovery of antibiotic substitutes, and has good application prospect in the aspect of preventing and treating the streptococcus suis.
Moreover, tea polyphenols have the following advantages: anticancer, antiaging, antioxidant, radioprotective, antibacterial, and antiviral effects. The tea polyphenols have protective effect on pigment and vitamins in food, so that the food can maintain original color and nutrition level for a long time, can effectively prevent food and edible oil from putrefaction, can eliminate peculiar smell, and lays a safety foundation for the food to be widely applied to the aspect of food storage in animal husbandry. In addition, the tea polyphenol has no toxicity to cells on a cellular level, has the characteristics of strong oxidation resistance, no toxic or side effect, no peculiar smell and the like, can be orally taken, has no stimulation, safety and no toxicity to organisms, has no three-effect, is expected to become a novel bioactive molecule for preventing and treating the streptococcus suis, provides a prerequisite for the research and development of antibacterial drugs of the tea polyphenol to the streptococcus suis, has a good application prospect in the aspect of preventing and treating the streptococcus suis, and provides a theoretical basis for the popularization and application of the tea polyphenol as an antibiotic substitute.
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FIG. 1 is a dynamic analysis of the growth curve of epigallocatechin gallate against growth of Streptococcus suis.
Detailed Description
The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available. The embodiment of the invention is illustrated by taking the streptococcus suis type 2 in the streptococcus suis as an example, and other streptococcus suis also has the inhibiting and killing effects on the substances of the invention.
Statistical analysis of the following examples of the invention: all experiments were repeated at least 3 times independently, with results expressed as mean and standard error, using one-way analysis of variance and T-test analysis. All statistical analyses used a P <0.05 as a test standard with significant statistical differences and the analysis software was GraphPad Prism 5.
Example 1:
determining the minimum inhibitory concentration and the minimum bactericidal concentration of the epigallocatechin gallate to the streptococcus suis:
1. material
Mueller-Hinton (MH) broth (from BD Co.) was used for drug sensitivity experiments to test the bacteriostatic efficacy of drugs.
2. Test method
Selecting a purified streptococcus suis type 2 SC19 bacterial colony by using a sterile inoculating loop, fully dispersing the bacterial colony in an MH liquid culture medium, and preparing a standard bacterial suspension with the concentration of 0.5 McLeod turbidimetric method;
epigallocatechin gallate solutions with different concentrations after being diluted by MH liquid culture medium in multiple ratios are respectively added into a sterile 96-well polystyrene plate, 50 mul of the epigallocatechin gallate solution is added into the 1 st to 10 th wells, 50 mul of the MH liquid culture medium is added into the 11 th well as a growth control, and 100 mul of the MH liquid culture medium is added into the 12 th well as a negative control.
After 0.5M/M.turbidimetric bacterial suspension was diluted 1: 1000 with MH broth, 50. mu.l of the bacterial suspension was added to wells 1 to 11, incubated at 37 ℃ for 16 to 20 hours, and the presence or absence of bacterial growth was observed to determine the MIC value. And sucking bacterial liquid from each hole, coating the bacterial liquid on an MH plate, and counting 5 colonies less to obtain an MBC value.
3. Results
As can be seen from Table 1, when the concentration of epigallocatechin gallate is 512 μ g/ml, it has inhibitory effect on growth of Streptococcus suis, and when the concentration of epigallocatechin gallate is 1024 μ g/ml, it has bactericidal effect on Streptococcus suis.
TABLE 1 minimum inhibitory and minimum bactericidal concentration of Epigallocatechin gallate on Streptococcus suis
Example 2:
inhibition of growth curve of streptococcus suis type 2 by epigallocatechin gallate:
the SC19 colony of purified Streptococcus suis type 2 was picked up by a sterile inoculating loop, dispersed in MH liquid medium, and a standard bacterial suspension with a concentration of 0.5 McLeod was prepared by McLeod's turbidimetry.
Epigallocatechin gallate solutions with different concentrations after diluted by MH liquid culture medium are respectively added into sterile bacteria bottles, and only MH liquid culture medium without the EGCG solution is used as a growth control.
And (3) equivalently inoculating 0.5 McLeod's turbidimetric standard bacterial suspension into the bacterial bottle, placing the bacterial bottle on a 37-degree shaking table for shake culture, taking bacterial liquid at intervals of two hours, diluting the bacterial liquid in a multiple ratio, coating an MH flat plate, and counting viable bacteria, wherein the result is expressed by CFU/ml.
As can be seen from FIG. 1, when the concentration of epigallocatechin gallate was 256. mu.g/ml (i.e., 0.5 MIC), there was no significant difference in the number of bacteria per time point as compared to the control group without epigallocatechin gallate. When the concentration of epigallocatechin gallate is 512 mug/ml (namely 1MIC value), the difference of the bacterial count at each time point is obvious compared with the control group without epigallocatechin gallate, which indicates that the concentration of epigallocatechin gallate is 512 mug/ml, and the antibacterial effect on streptococcus suis is achieved. When the concentration of epigallocatechin gallate was 1024. mu.g/ml (i.e., 2MIC value), Streptococcus suis was killed by the second hour of the culture.
The results show that the epigallocatechin gallate can not only inhibit the growth of the streptococcus suis type 2, but also kill the streptococcus suis type 2.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (3)
1. Application of epigallocatechin gallate in preparing medicine for treating or preventing Streptococcus suis type 2 infection is provided.
2. The use according to claim 1, wherein the epigallocatechin gallate is used for inhibiting the growth of Streptococcus suis type 2 in vitro.
3. The use of claim 1, wherein the epigallocatechin gallate is used for killing Streptococcus suis type 2 in vitro.
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| CN111249264B (en) * | 2020-02-18 | 2023-07-07 | 东北农业大学 | Use of gallic acid in reversing resistance of streptococcus suis to antibiotics |
| CN114288287A (en) * | 2022-01-11 | 2022-04-08 | 湖北省农业科学院畜牧兽医研究所 | Application of theaflavin in preparation of anti-streptococcus suis medicine |
| CN116139143A (en) * | 2022-09-07 | 2023-05-23 | 山东省妇幼保健院 | Application of natural medicine in preparation of medicine for resisting gram-positive bacteria |
| CN115317499B (en) * | 2022-09-21 | 2023-08-04 | 湖北省农业科学院畜牧兽医研究所 | Application of timosaponin AIII in preparation of drug for inhibiting haemophilus parasuis |
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| CN101940574A (en) * | 2010-07-23 | 2011-01-12 | 中国人民解放军第三军医大学 | Application of combination of combined catechin substances with antibacterial drugs |
| CN106667997A (en) * | 2017-02-27 | 2017-05-17 | 吉林大学 | EGCG with inhibitory effect on streptococcus pneumoniae pathogenicity and application |
| CN109674785A (en) * | 2019-01-18 | 2019-04-26 | 南方医科大学 | Epigallo-catechin gallate (EGCG) and its derivative are preparing the application in anti-zika virus drug |
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Non-Patent Citations (3)
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| 儿茶素类化合物抑菌作用及其作用机制的研究进展;陈书媛等;《食品安全质量检测学报》;20170915;第8卷(第9期);3316-3322 * |
| 茶多酚抗菌作用研究进展;田福忠等;《现代化农业》;20200515(第5期);29-32 * |
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