Application of citral in inhibition of growth of multidrug-resistant providencia rettgeri
阅读说明：本技术 柠檬醛在抑制多重耐药雷氏普罗威登斯菌生长中的应用 (Application of citral in inhibition of growth of multidrug-resistant providencia rettgeri ) 是由 钱卫东 付玉婷 刘淼 李靖原 王文静 李鑫辰 黄杰 刘婉婷 于 2019-12-04 设计创作，主要内容包括：本发明公开了柠檬醛在抑制多重耐药雷氏普罗威登斯菌生长中的应用,根据柠檬醛对耐氨苄西林、氨苄西林/舒巴坦、头孢唑林、头孢替坦、头孢曲松、头孢他啶、妥布霉素、哌拉西林他唑巴坦、环丙沙星、左氧氟沙星、阿米卡星、复方新诺明、头孢吡肟、厄他培南、亚胺培南、氨曲南、庆大霉素的人源雷氏普罗威登斯菌,具有较好的体外杀灭作用,能够抑制耐雷氏普罗威登斯菌的生长,且最低杀菌浓度为0.5mg/mL,最小抑菌浓度为0.25mg/mL；本发明提供植物提取物柠檬醛对多重耐药雷氏普罗威登斯菌的抑菌作用,可缓解或解决多重耐药雷氏普罗威登斯菌的耐药性增加问题,为临床抗细菌治疗及新型药物研发提供理论依据。(The invention discloses an application of citral in inhibiting the growth of multidrug-resistant providencia rettgeri, wherein the human providencia rettgeri which is resistant to ampicillin, ampicillin/sulbactam, cefazolin, cefotetan, ceftriaxone, ceftazidime, tobramycin, piperacillin tazobactam, ciprofloxacin, levofloxacin, amikacin, compound sulfamethoxazole, cefepime, ertapenem, imipenem, aztreonam and gentamycin has a better in-vitro killing effect according to the citral, the growth of the providencia rettgeri can be inhibited, the minimum bactericidal concentration is 0.5mg/mL, and the minimum bacteriostatic concentration is 0.25 mg/mL; the invention provides the bacteriostatic action of the plant extract citral on the multidrug-resistant providencia rettgeri, can relieve or solve the problem of drug resistance increase of the multidrug-resistant providencia rettgeri, and provides a theoretical basis for clinical antibacterial treatment and novel drug research and development.)
1. Application of citral in inhibiting growth of multiple drug-resistant providencia rettgeri.
2. The use of citral according to claim 1 for inhibiting the growth of multidrug-resistant providencia rettgeri, wherein the multidrug-resistant providencia rettgeri is human providencia rettgeri.
3. The use of citral according to claim 1 for inhibiting the growth of multidrug-resistant providencia rettgeri, wherein the resistance of providencia rettgeri is determined by microdilution.
4. Use of citral according to claim 1 for inhibiting the growth of multidrug resistant providencia rettgeri, wherein the multidrug resistant providencia rettgeri resistant antibiotic is selected from the group consisting of ampicillin, ampicillin/sulbactam, cefazolin, cefotetan, ceftriaxone, ceftazidime, tobramycin, piperacillin tazobactam, ciprofloxacin, levofloxacin, amikacin, sulfamethoxazole, cefepime, ertapenem, imipenem, nitrofurantoin, aztreonam, gentamicin.
5. Use of citral according to claim 1 for inhibiting the growth of multidrug resistant providencia rettgeri selected from the group consisting of ampicillin resistant, ampicillin/sulbactam, cefazolin, cefotetan, ceftriaxone, ceftazidime, tobramycin, piperacillin tazobactam, ciprofloxacin, levofloxacin, amikacin, ziram, cefepime, ertapenem, imipenem, aztreonam, gentamicin human providencia rettgeri.
6. The use of citral according to claim 5 for inhibiting the growth of multidrug-resistant providencia rettgeri, wherein the citral has a minimum inhibitory concentration of 0.25mg/mL and a minimum bactericidal concentration of 0.5mg/mL against the human providencia rettgeri.
7. The use of citral according to claim 1 for inhibiting the growth of multidrug-resistant providencia rettgeri, wherein the citral has an in vitro killing effect on the multidrug-resistant providencia rettgeri and inhibits the in vitro growth of the multidrug-resistant providencia rettgeri.
8. Application of citral in preparation of multiple drug resistant providencia rettgeri resistant drugs.
9. Use of citral according to claim 8 for the preparation of a medicament against multidrug-resistant providencia rettgeri, wherein the multidrug-resistant providencia rettgeri is human providencia rettgeri.
10. Use of citral according to claim 8 for the preparation of a medicament against multidrug resistant providencia rettgeri selected from the group consisting of multiple antibiotic resistant providencia rettgeri consisting of ampicillin, ampicillin/sulbactam, cefazolin, cefotetan, ceftriaxone, ceftazidime, tobramycin, piperacillin tazobactam, ciprofloxacin, levofloxacin, amikacin, sulfamethoxazole, cefepime, ertapenem, imipenem, aztreonam, gentamicin.
Technical Field
The invention belongs to the technical field of medicine and food safety, and relates to application of citral in inhibition of growth of multidrug-resistant providencia rettgeri.
Background
The providencia rettgeri enterobacteriaceae can cause urinary tract infection and other parenteral infections, and cause outbreaks of a plurality of nosocomial infections, and is also a food companion bacterium which is extremely easy to propagate in large quantities in the processes of food processing, transportation, sale and the like to cause intestinal infection and food poisoning.
Tolerance and detection rates of multidrug-resistant providencia rettgeri are also increasing due to clinical abuse of antibiotics in many countries. In the face of the problem of enhancing the drug resistance of bacteria, the difficulty of developing a new drug with antibacterial activity by using a traditional chemical synthesis method is long, Chinese herbal medicines are important sources for new drug discovery, the natural products of the new drug are complex and diverse in structure and category and have various pharmacological activities such as antibacterial activity and anti-inflammatory activity, and natural plant essential oil is expected to become a preferable natural substitute of a chemical synthesis antibacterial agent due to the advantages of safety, low toxicity, broad-spectrum antibacterial activity, no drug resistance and the like.
Citral (Citral) is an important monoterpene compound, exists in various aromatic plant essential oils such as citrus oil, lemon oil, lemongrass oil, pungent litse fruit oil, etc., has unique lemon flavor, and is a common raw material in the fields of spice, food and pharmacy. Previous studies have shown that citral has biological activities such as insecticidal, repellent, antibacterial, antioxidant, etc. (Liu Yan, Su shou, Chen Shang \38040, Chen jin Zhu, Van Guo Rong. the research on the biological activity of citral has been advanced [ J]Jiangxi forestry science and technology, 2013(01): 43-46.). The document (research on antibacterial mechanism of Escherichia coli O157H 7 by using Dajinming litsea cubeba essential oil and application thereof in fruit and vegetable juice [ D]University of Jiangsu, 2019.) states that citral has an intercalation binding effect with DNA of Escherichia coli O157: H7, destroying the DNA structure, and thus has a significant antibacterial effect against Escherichia coli. In addition, Mesquita et al reported that citral has antibacterial activity against Staphylococcus aureus, has a MIC of 0.5mg/mL, and has the potential to clear biofilms. (Porf i rio Emanuula Mesquita, Melo Hider Machado, Pereira)Matheus Gomes,Cavalcante TheodoraThays Arruda,Gomes Geovany Amorim,de Carvalho Mário Geraldo,Costa RenataAlbuquerque,Júnior Francisco Eduardo
Disclosure of Invention
The invention aims to provide application of citral in inhibiting the growth of multidrug-resistant providencia rettgeri, and explores potential drug-resistant bacteria inhibitors from the existing medicinal plant resource library, so as to provide a practical foundation for developing novel antibiotic substitutes and solving the multidrug-resistant predicament of drug-resistant enterobacter cloacae.
The invention is realized by the following technical scheme:
application of citral in inhibiting growth of multiple drug-resistant providencia rettgeri.
Further, the multidrug-resistant providencia rettgeri is human providencia rettgeri.
Further, the resistance to providencia rettgeri was determined by the microdilution method.
Further, the antibiotic resistant to multiple drug resistant providencia rettgeri is selected from a plurality of ampicillin, ampicillin/sulbactam, ceftizolin, cefotetan, ceftriaxone, ceftazidime, tobramycin, piperacillin tazobactam, ciprofloxacin, levofloxacin, amikacin, sulfamethoxazole, cefepime, ertapenem, imipenem, nitrofurantoin, aztreonam, gentamicin.
Further, the multidrug-resistant providencia rettgeri is selected from human providencia rettgeri having ampicillin resistance, ampicillin/sulbactam resistance, cefazolin resistance, cefotetan resistance, ceftriaxone resistance, ceftazidime resistance, tobramycin resistance, piperacillin tazobactam resistance, ciprofloxacin resistance, levofloxacin resistance, amikacin resistance, compound sulfamethoxine resistance, cefepime resistance, ertapenem resistance, imipenem resistance, aztreonam resistance and gentamicin resistance.
Further, the minimum inhibitory concentration of the citral on the human providencia rettgeri is 0.25mg/mL, and the minimum bactericidal concentration is 0.5 mg/mL.
Further, citral has an in vitro killing effect on multiple drug-resistant providencia rettgeri and inhibits the in vitro growth of multiple drug-resistant providencia rettgeri.
Application of citral in preparation of multiple drug resistant providencia rettgeri resistant drugs.
Further, the multidrug-resistant providencia rettgeri is human providencia rettgeri.
Further, the multiple drug resistant providencia rettgeri is selected from the group consisting of multiple antibiotic resistant providencia rettgeri of ampicillin, ampicillin/sulbactam, ceftizolid, cefotetan, ceftriaxone, ceftazidime, tobramycin, piperacillin tazobactam, ciprofloxacin, levofloxacin, amikacin, sulfamethoxazole, cefepime, ertapenem, imipenem, aztreonam, gentamicin.
Compared with the prior art, the invention has the following beneficial technical effects:
based on the research of the effect of citral on the multiple drug-resistant providencia rettgeri, the citral is used for inhibiting the growth of the multiple drug-resistant providencia rettgeri, so that a new idea is provided for the research, development and application of the multiple drug-resistant providencia rettgeri inhibitor, and the citral has a wide application value in the field of medicines.
The invention further defines the inhibition effect of citral on the drug resistance of human providencia retta of antibiotic-resistant ampicillin, ampicillin/sulbactam, cefazolin, cefotetan, ceftriaxone, ceftazidime, tobramycin, piperacillin tazobactam, ciprofloxacin, levofloxacin, amikacin, compound sulfamethoxazole, cefepime, ertapenem, imipenem, nitrofurantoin, aztreonam and gentamicin, effectively relieves or solves the problem of drug-resistant infection of multiple drug-resistant providencia retta bacteria, and reduces the mortality.
Detailed Description
The present invention will be described in further detail with reference to examples. The examples are only for explaining the contents of the present invention and do not limit the present invention.
Citral, British name Citral, molecular formula C10H16O, 152.23 relative molecular mass, is a colorless or yellowish liquid with a strong lemon flavor. Is contained in plant essential oil such as citrus oil, lemon oil, lemongrass oil, pungent litse fruit oil, white lemon oil, and verbena oil. CAS number 5392-40-5, boiling point 228-:
1. drug susceptibility test of providencia rettgeri
The invention takes 5 strains of providencia rettgeri as starting strains, and selects 18 common antibiotics such as ampicillin, ampicillin/sulbactam, ceftizoxime, cefotetan, ceftriaxone, ceftazidime, tobramycin, piperacillin tazobactam, ciprofloxacin, levofloxacin, amikacin, compound sulfamethoxine, cefepime, ertapenem, imipenem, nitrofurantoin, aztreonam, gentamicin and the like to carry out drug susceptibility experiments.
The pure colonies cultured for 16h are picked up and uniformly dissolved in 3mL of sterile physiological saline, and the turbidity of the pure colonies is adjusted to be equal to that of a 0.5 McLeeb turbiditube. Adding antibiotics, bacterial liquid and TSB liquid culture medium into a 96-hole culture plate by using a two-fold dilution method for overnight culture, and measuring the minimum inhibitory concentration of the antibiotics to providencia rettgeri by using a microplate reader. Three liquid medicine groups with different concentrations are parallel to ensure the reliability of experimental data. The bacteriostatic result is judged according to the national standard administration committee of the united states clinical laboratory (CLSl2017), and the judgment standard is shown in table 1. The results are shown in Table 2 and the results show that the number is 1#The providencia rettgeri can tolerate the most antibiotics, can tolerate 17 antibiotics and is sensitive to 1 antibiotic, so 1 antibiotic is required#As a further experimental study.
TABLE 1 results of the national Committee for standardization management of the clinical laboratory (CLSl2017) standards
TABLE 2 results of antibiotic versus minimum inhibitory concentration of providencia rettgeri
As can be seen from tables 1 and 2, citral has a good inhibitory effect on multiple drug-resistant providencia retta bacteria of ampicillin, ampicillin/sulbactam, ceftizone, cefotaxime, ceftriaxone, ceftazidime, tobramycin, piperacillin tazobactam, ciprofloxacin, levofloxacin, amikacin, sulfamethoxazole, cefepime, ertapenem, imipenem, aztreonam, gentamicin.
2. Inhibition of multi-drug resistant strains by citral
In order to fully consider the medication safety, the invention takes the single active component of citral as a research object, takes the standard strain ATCC31052 of providencia rettgeri as a reference, and carries out the research of drug resistance inhibition. The pure colonies cultured for 16h are picked and uniformly dissolved in 3mL LB liquid medium, the turbidity is adjusted to be 0.5 McLeod, and the OD600 value is measured by an enzyme-labeling instrument. The citral with the concentration of 2mg/mL is prepared by dimethyl sulfoxide and used as a liquid medicine, the liquid medicine, a bacterial liquid and an LB liquid culture medium are added into a 96-hole culture plate by using a test tube double dilution method for overnight culture, and three parallel experiments are carried out by using liquid medicine groups with different concentrations to ensure the reliability of experimental data.
And (3) determining the minimum inhibitory concentration of citral to the multiple drug-resistant providencia rettgeri by using an enzyme labeling instrument. And placing the culture solution under the minimum inhibitory concentration in a sterile LB liquid culture medium for continuous culture, and determining the minimum bactericidal concentration of citral on the multi-drug-resistant providencia rettgeri by using an enzyme labeling instrument, wherein the experimental results are shown in Table 3.
TABLE 3 inhibition of Multi-drug resistant providencia rettgeri by citral
As shown in Table 3, the MIC of citral against multidrug-resistant providencia rettgeri was 0.25mg/mL, and the MBC was 0.5 mg/mL.
According to the experimental results, the drug resistance characteristics of drug-resistant pathogenic bacteria are solved by combining Chinese herbal medicines, the active single-product component citral can be obtained, the clinical main multiple-drug-resistant providencia rettgeri can be directly inhibited, the drug-resistant infection problem of the multiple-drug-resistant providencia rettgeri can be effectively relieved or solved, a new thought is provided for inhibiting the multiple-drug-resistant providencia rettgeri, and the method has important guiding significance.
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