CN110946862B - Application of sanguinarine in inhibition of growth of multiple drug-resistant enterobacter hopcalis - Google Patents
Application of sanguinarine in inhibition of growth of multiple drug-resistant enterobacter hopcalis Download PDFInfo
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- CN110946862B CN110946862B CN202010005995.6A CN202010005995A CN110946862B CN 110946862 B CN110946862 B CN 110946862B CN 202010005995 A CN202010005995 A CN 202010005995A CN 110946862 B CN110946862 B CN 110946862B
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- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
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- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4741—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having oxygen as a ring hetero atom, e.g. tubocuraran derivatives, noscapine, bicuculline
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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 discloses an application of sanguinarine in inhibiting the growth of multidrug-resistant enterobacter hopcalis, which has a good in-vitro killing effect on the multidrug-resistant enterobacter hopcalis with piperacillin resistance, levofloxacin resistance, bromgmen resistance, imipenem resistance, ofloxacin resistance, nitrofurantoin resistance, cefotaxime resistance, azithromycin resistance and nalidixic acid resistance, can inhibit the growth of the multidrug-resistant enterobacter hopcalis, and has a minimum inhibitory concentration of 0.12mg/mL and a minimum bactericidal concentration of 0.24 mg/mL.
Description
Technical Field
The invention relates to application of a traditional Chinese medicine effective component sanguinarine as an inhibitor of enterobacter, in particular to application of sanguinarine in inhibition of growth of enterobacter hophallii.
Background
Enterobacter hollisae (Enterobacter hormaechei) is a gram-negative Enterobacter (Enterobacter of Enterobacteriaceae) which is mainly derived from human, animals and food, wherein the food source comprises dairy products, dried meat, milk, fruits, vegetables, rice and the like, and the Enterobacter hollisae is also separated from infant milk powder, and a learner (Cheng Jiang, Von Asian, Wangpong, et al. separation identification and drug sensitivity analysis of several pathogenic bacteria causing the lung disease of the forest musk deer [ J ]. Chinese veterinary journal (12):78-82.) verifies that the Enterobacter hollisae has certain conditional pathogenicity according to a mouse infection test. And it has also been reported that infection with Enterobacter hollisae can also cause bacteremia including stroke, premature birth and weak infants. The enterobacter hopcalis is a main pathogenic bacterium causing intestinal infectious diseases of human or animals, and can cause a series of gastrointestinal diseases such as diarrhea, enteritis, gastrointestinal infection, colorectal cancer, colon cancer and the like. It can be seen that the source, pathogenicity, drug resistance and potential harm of the enterobacter hopcalis are increasingly aggravated.
Bacterial resistance to antibiotics, especially multidrug resistance, has become a medical and social concern worldwide, seriously threatening the treatment of infectious diseases. These bacteria include "superbacteria," which have evolved to produce multiple resistance mechanisms to the action of antibiotics due to the diversity and mobility of their own genetic structures. The high resistance presented by resistant bacteria to a wide variety of different chemical structure antibiotics makes selection of anti-infective therapeutic drugs extremely difficult.
In the face of the current situation of severe antibiotic resistance, the idea of non-antibiotic treatment of infectious diseases is gradually agreed and studied with great success. The Chinese herbal medicine also has obvious effects on inhibiting the growth of bacteria and killing pathogenic bacteria. Clinical verification shows that many medicinal materials have very good antibacterial and anti-inflammatory effects, and have the characteristics of small toxic and side effects and difficulty in generating drug resistance. Sanguinarine is a benzophenanthroline alkaloid existing in natural plants and has great development prospect. The molecular formula is C20H14NO4The molecular weight is 332.33, the CAS number is 2447-54-3, and the melting point is 277-280 ℃. Sanguinarine is mainly present in natural medicinal plants of Papaveraceae and Rutaceae, and the safety, function and application of sanguinarine (gentamicin, Xia-terraning, Hu Rui Zhi, et al.) [ J ] are generally studied]Feed Explorer, 2018, No.315(07):49-53.) reported that sanguinarine has bactericidal activityAntibacterial, anti-inflammatory, analgesic, anti-tumor, smooth muscle contraction inhibition and insecticidal effects, and can be used as feed additive in livestock and poultry production, and can promote animal growth, improve animal meat quality, improve intestinal tract and enhance immunity, and also can be used as literature (Hamoud R, Reichling, Jurgen, wine M. synthetic antibiotic activity of the combination of the alkyl and the antibiotic tissue with EDTA and the antibiotic tissue against antibiotic vaccine bacteria [ J]Journal of Pharmacy and Pharmacology 2015,67(2): 264-. However, sanguinarine is reported as an active bacteriostatic component for inhibiting the growth of multidrug-resistant enterobacter hophallus.
Disclosure of Invention
The invention aims to solve the problem of drug resistance in current clinical medicine and food safety, and aims to provide the application of sanguinarine in inhibiting the growth of multiple-drug-resistant enterobacter hophallii.
In order to achieve the purpose, the invention adopts the technical scheme that:
the drug resistance of the enterobacter hopcalis to antibiotics is determined by a paper diffusion method, and the Minimum Inhibitory Concentration (MIC) and the Minimum Bactericidal Concentration (MBC) of sanguinarine to the enterobacter hopcalis are determined by a micro-double dilution method. The results show that: the sanguinarine has good bacteriostatic action on the multiple drug-resistant enterobacter hopcalis, and can be used for inhibiting the growth of the multiple drug-resistant enterobacter hopcalis.
Preferably, the multidrug-resistant enterobacter hopcalis is human-derived or food-derived enterobacter hopcalis resistant to multiple antibiotics in piperacillin, levofloxacin, bromogeltin, imipenem, ofloxacin, nitrofurantoin, cefotaxime, azithromycin and nalidixic acid.
Preferably, the sanguinarine has an inhibitory effect on human-derived or food-derived enterobacter hopcalis resistant to piperacillin, levofloxacin, meglumine, imipenem, ofloxacin, nitrofurantoin, cefotaxime, azithromycin and nalidixic acid.
Preferably, the minimum inhibitory concentration of sanguinarine is 0.12mg/mL, and the minimum bactericidal concentration is 0.24 mg/mL.
The invention has the beneficial effects that:
the invention starts from the existing medicinal plant resource library, excavates potential drug-resistant bacteria inhibitor, finds that the growth of multiple drug-resistant enterobacter hopcalis can be effectively inhibited by utilizing non-antibiotic compound sanguinarine based on the research on the effect of sanguinarine on multiple drug-resistant enterobacter hopcalis, provides new thought and source for the research, development and application of multiple drug-resistant enterobacter hopcalis inhibitor, and has wide application value in the fields of food, medicine and the like.
Furthermore, the invention defines the inhibition effect of sanguinarine on multiple drug-resistant enterobacter hopcalis, and the sanguinarine can be used as a substitute of antibiotics, so that the drug-resistant and infection problems of multiple drug-resistant enterobacter hopcalis can be effectively relieved or solved, and the fatality rate is reduced.
Detailed Description
The present invention will be described in further detail with reference to examples. The examples are only for explaining the present invention and do not limit the scope of protection of the present invention.
1. Drug sensitivity test of enterobacter hopcalis
The invention takes multi-strain human-derived enterobacter hopcalis (a strain sample is taken from a certain Beijing hospital) as an original strain, and selects drug sensitive test paper sheets of 10 common antibiotics such as piperacillin, levofloxacin, gentamicin, bromgmelin, imipenem, ofloxacin, nitrofurantoin, cefotiaquine, azithromycin and nalidixic acid to carry out drug sensitive test.
And (3) selecting and culturing the single bacterial colony for 18-24 hours, uniformly dissolving the single bacterial colony in 2-5 mL of sterile physiological saline, and adjusting the turbidity of the single bacterial colony to be equal to that of a 0.5 McLee turbidimetric tube. Sucking 50-200 mu L of bacterial liquid under aseptic conditions, uniformly coating the bacterial liquid on a flat plate by using a cotton swab, and standing at room temperature for 3-5 min. Then, the drug sensitive test paper (diameter 6mm) is tightly attached to the surface of the flat plate by using sterile forceps as required, and three antibiotic drug sensitive test paper sheets are taken to be parallel. And placing the inoculated flat plate in an incubator at 36-37 ℃ for incubation for 16-24 h. The diameter of the inhibition zone is measured by a vernier caliper with the precision of 0.02mm, the sensitivity of the enterobacter heuchei to antibiotics is judged according to the diameter of the inhibition zone, the inhibition result is judged according to the standard of the national standard administration committee (CLSl2017) of the American clinical laboratory, and the judgment standard is shown in the table 1. The experimental results are shown in table 2, and the results show that 1 strain of enterobacter hopcalis (strain # 1) can tolerate 9 clinically common antibiotics and is only sensitive to gentamicin, so that the strain is taken as a research object of the next experiment.
TABLE 1 results of the national Committee for standardization management of the clinical laboratory (CLSl2017) standards
TABLE 2 diameter results of zone of inhibition in drug susceptibility test of human Enterobacter Huoshanense
Note: the unit of the diameter of the inhibition zone is mm, and the numerical values are the average values of three parallels
2. Inhibition of multiple drug-resistant strains by sanguinarine
In order to fully consider the medication safety, the invention takes sanguinarine which is a single active monomer component as a research object, takes ATCC700323 as a reference strain (the reference strain is purchased from the North Na biological strain collection center, and is highly sensitive to imipenem and meropenem), and takes the No. 1 strain as an experimental strain to research the drug resistance inhibition effect. Selecting and culturing single bacterial colony for 18-24 h, and uniformly dissolving the single bacterial colony in 2-5 mL of TSB solutionAdjusting turbidity of the culture medium to 0.5 McLeod turbidimetric tube, and measuring OD with enzyme-labeling instrument600The value is obtained. The sanguinarine with the concentration of 50mg/mL is prepared by dimethyl sulfoxide and used as a liquid medicine, the liquid medicine, a bacterial liquid and a TSB liquid culture medium are added into a 96-hole culture plate by a test tube double dilution method for culturing for 24 hours, and liquid medicine groups with different concentrations are all three parallel, so that the reliability of experimental data is ensured.
And (3) determining the Minimum Inhibitory Concentration (MIC) of sanguinarine to the multidrug-resistant enterobacter hophallii by using a microplate reader. And transferring the culture solution with the concentration of the MIC diluted by the former drug to a sterile TSA solid culture medium for further culture for 24h, observing whether a single colony is generated, and if not, determining the concentration as the Minimum Bactericidal Concentration (MBC) of the multidrug-resistant Enterobacter huoshimi, wherein the experimental results are shown in Table 3.
TABLE 3 inhibition of multiple drug-resistant Enterobacter hollisae by sanguinarine
As shown in Table 3, sanguinarine has a good inhibitory effect on multidrug-resistant Enterobacter holdii of piperacillin resistance, levofloxacin resistance, bromine glutenin resistance, imipenem resistance, ofloxacin resistance, nitrofurantoin resistance, cefotaxime resistance, azithromycin resistance and nalidixic acid resistance, the MIC of the sanguinarine is 0.12mg/mL, and the MBC of the sanguinarine resistance is 0.24 mg/mL. Moreover, the result of comparison with the reference strain ATCC700323 shows that sanguinarine not only has certain bacteriostatic action on the reference strain ATCC700323, but also has certain bacteriostatic action on the multiple drug-resistant Enterobacter hollisae (such as the No. 1 strain) in the invention.
According to the experimental results, the characteristics of wide sources of Chinese herbal medicines, less adverse reactions, difficult generation of drug resistance and the like are integrated, sanguinarine active monomer components can be obtained by extracting and separating the Chinese herbal medicines, the inhibition effect is directly exerted on the multiple drug-resistant enterobacter hopcalis, clinical diseases caused by the enterobacter hopcalis can be effectively treated, the infection problem of the multiple drug-resistant enterobacter hopcalis can be effectively relieved or solved, the mortality rate of diseases is reduced, scientific basis is provided for researching bacteriostat of the clinically separated enterobacter hopcalis, and new ideas and sources are provided for developing drugs and antibiotic substitutes for inhibiting the multiple drug-resistant enterobacter hopcalis.
Claims (5)
1. Application of sanguinarine in preparing medicine for resisting multiple drug-resistant Enterobacter holtzeri is provided.
2. Use according to claim 1, characterized in that: the multidrug-resistant enterobacter hopcalis is human-derived or food-derived enterobacter hopcalis.
3. Use according to claim 1, characterized in that: the multiple drug-resistant enterobacter hopcalis is selected from enterobacter hopcalis which is resistant to multiple antibiotics in piperacillin, levofloxacin, bromgeltin, imipenem, ofloxacin, nitrofurantoin, cefotaxime, azithromycin and nalidixic acid.
4. Use according to claim 3, characterized in that: the minimum inhibitory concentration of the sanguinarine to the multidrug-resistant enterobacter hophallus is 0.12mg/mL, and the minimum bactericidal concentration is 0.24 mg/mL.
5. Use according to claim 1, characterized in that: the sanguinarine has in vitro bacteriostatic and bactericidal effects on multiple drug-resistant enterobacter hopcalis.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110279701A (en) * | 2019-07-16 | 2019-09-27 | 陕西科技大学 | Ursolic acid is inhibiting the application in the growth of multidrug resistant enterobacter cloacae |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110279701A (en) * | 2019-07-16 | 2019-09-27 | 陕西科技大学 | Ursolic acid is inhibiting the application in the growth of multidrug resistant enterobacter cloacae |
Non-Patent Citations (4)
| Title |
|---|
| Structural modification of sanguinarine and chelerythrine and their antibacterial activity;Fang Miao等;《Natural Product Research》;20110531;第25卷(第9期);第863-875页 * |
| Structure-Activity Relationship of Benzophenanthridine Alkaloids from Zanthoxylum rhoifolium Having Antimicrobial Activity;Luciana de C. Tavares等;《PLoS One》;20140531;第9卷(第5期);第1-10页 * |
| Synergistic antibacterial activity of the combination of the alkaloid sanguinarine with EDTA and the antibiotic streptomycin against multidrug resistant bacteria;Razan Hamoud等;《Journal of Pharmacy and Pharmacology》;20141231;第1-10页 * |
| 茶籽多糖和博落回生物碱对黄羽肉鸡肠道菌群的影响及博落回血根碱的抑菌活性;胡贵丽等;《动物营养学报》;20181231;第30卷(第11期);第4619-4625页 * |
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