CN111728979A - Application of baohuogan I in resisting staphylococcus aureus biofilm infection - Google Patents
Application of baohuogan I in resisting staphylococcus aureus biofilm infection Download PDFInfo
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- CN111728979A CN111728979A CN202010812651.6A CN202010812651A CN111728979A CN 111728979 A CN111728979 A CN 111728979A CN 202010812651 A CN202010812651 A CN 202010812651A CN 111728979 A CN111728979 A CN 111728979A
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- baohuogan
- staphylococcus aureus
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- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
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Abstract
The invention discloses an application of baohuogan I in resisting staphylococcus aureus biofilm infection. The technical scheme of the invention discloses a new application of baohuogan I, which is used for resisting staphylococcus aureus biofilm infection, and in an environment infected by staphylococcus aureus biofilm, baohuogan I can effectively inhibit the formation of staphylococcus aureus biofilm and can be used together with other antibiotics to effectively remove the formed staphylococcus aureus biofilm.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to application of baohuogan I in resisting staphylococcus aureus biofilm infection.
Background
Staphylococcus aureus can infect different parts of the human body to cause various infectious diseases, from common skin diseases such as folliculitis, acne and hordeolum to deep and fatal diseases such as pneumonia, endocarditis, osteomyelitis and other metastatic complications. The staphylococcus aureus can be adhered to the surface of human tissue cells or medical implant materials to form a biofilm structure consisting of extracellular polysaccharide adhesion molecules, proteins, teichoic acid, extracellular DNA (eDNA) and the like, so that the sensitivity of bacteria to antibacterial drugs is reduced, and the attack and phagocytosis of host immune cells are avoided, thereby causing chronic infection and delayed recovery. The serious problem is that the infection in hospital related to staphylococcus aureus biofilm is increasing with the wide application of medical implant materials such as various catheters, dialysis technology, prosthetic joints and the like in recent years. At present, a few medicines for resisting staphylococcus aureus biofilm infection are available, and only clindamycin, azithromycin, linezolid, daptomycin and the like have certain treatment effects, but the treatment effect on biofilm infection of partial strains is poor. Therefore, the discovery of drugs having therapeutic effects against staphylococcus aureus biofilm infection is one of the current research hotspots.
Baohuoside i (baohuoside i) is a flavonoid compound obtained from epimedium herb, and has pharmacological actions of resisting tumors, treating osteoporosis and improving cognitive dysfunction. At present, no research report on related antibacterial performance exists in baohuogan I.
Disclosure of Invention
Aiming at the technical problems, the invention discloses application of baohuogan I in resisting staphylococcus aureus biofilm infection, the application is a new application of baohuogan I, and the baohuogan I can obviously inhibit the formation of staphylococcus aureus biofilm and can be combined with other antibiotics to effectively remove the formed staphylococcus aureus biofilm.
In contrast, the technical scheme adopted by the invention is as follows:
use of baohuogan I for resisting staphylococcus aureus biofilm infection, and the baohuogan I for resisting staphylococcus aureus biofilm infection. At present, baohuogan I is used as a medicament for resisting tumors, treating osteoporosis and improving cognitive dysfunction. The research shows that the baohuogan I also has the application of resisting the infection of the staphylococcus aureus biofilm, can be used in the medicines for resisting the infection of the staphylococcus aureus biofilm, inhibits the formation of the staphylococcus aureus biofilm, and can be used together with other antibiotics to effectively remove the formed staphylococcus aureus biofilm.
As a further improvement of the invention, baohuogan I is used for resisting staphylococcus aureus biofilm infection.
As a further improvement of the invention, the concentration of the baohuogan I is 6.25 to 100 mu M.
As a further improvement of the invention, the concentration of baohuogan I is not less than 12.5 mu M.
As a further improvement of the invention, the concentration of baohuogan I is not less than 25 mu M.
The invention discloses an application of baohuogan I in preparation of a medicine for resisting staphylococcus aureus infection.
Furthermore, in the anti-staphylococcus aureus infection medicine, the concentration of baohuogan I is less than or equal to 100 MuM. Further, the medicine can be in the form of spray, oral granules, oral tablets or injection.
As a further improvement of the invention, the concentration of baohuogan I is not less than 12.5 mu M.
As a further improvement of the invention, the concentration of baohuogan I is not less than 25 mu M. The baohuogan I with the concentration of 25 mu M has extremely strong activity of inhibiting the formation of the biofilm.
As a further improvement of the invention, the medicament against Staphylococcus aureus infection comprises other medicaments, such as other antibiotics. Further, the anti-staphylococcus aureus drug comprises daptomycin. The baohuogan I and daptomycin are combined, so that the effect of resisting staphylococcus aureus infection is better.
As a further improvement of the invention, the effective concentration range of the baohuogan I for clearing the formed biofilm of staphylococcus aureus by combining with other medicines is 1.56-100 mu M.
As a further improvement of the invention, the concentration of baohuogan I is not less than 12.5 mu M.
As a further improvement of the invention, the concentration of baohuogan I is not less than 50 μ M.
As a further improvement of the invention, the concentration of the baohuogan I is less than or equal to 100 mu M.
The invention also discloses an application of baohuogan I in preparing a coating for the surface of a medical instrument, wherein the coating for the surface of the medical instrument contains baohuogan I. The baohuogan I is used for inhibiting the formation of a staphylococcus aureus biofilm and the adhesion of staphylococcus aureus.
As a further improvement of the invention, in the coating for the surface of the medical device, the concentration of baohuogan I is less than or equal to 100 MuM. Preferably, the concentration of baohuogan I is 6.25 to 100 μ M. Preferably, the concentration of baohuogan I is not less than 12.5 mu M. Preferably, the concentration of baohuogan I is not less than 25 μ M.
The concentration of baohuogan I is 6.25 mu M-100 mu M, and the baohuogan I has the capability of inhibiting the formation of a biofilm, so that the adhesion of staphylococcus aureus on the surface of a medical material can be reduced if the compound is directly adhered to the surface of the medical material.
The invention also discloses a coating for the surface of medical equipment, which comprises baohuogan I. Preferably, the concentration of baohuogan I is 6.25 to 100 μ M. Preferably, the concentration of baohuogan I is not less than 12.5 mu M. Preferably, the concentration of baohuogan I is not less than 25 μ M. Preferably, the concentration of baohuogan I is less than or equal to 100 mu M.
The invention also discloses an application of the baohuogan I in preparing a disinfectant for resisting staphylococcus aureus, wherein the disinfectant comprises the baohuogan I which is used for inhibiting the formation of a biofilm of the staphylococcus aureus and the adhesion of the staphylococcus aureus.
The invention also discloses a disinfectant for resisting staphylococcus aureus, which comprises baohuogan I. Preferably, the concentration of baohuogan I is 6.25 to 100 μ M. Preferably, the concentration of baohuogan I is not less than 12.5 mu M. Preferably, the concentration of baohuogan I is not less than 25 μ M. Preferably, the concentration of baohuogan I is less than or equal to 100 mu M.
Compared with the prior art, the invention has the beneficial effects that:
the technical scheme of the invention discloses a new application of baohuogan I, which is used for resisting staphylococcus aureus biofilm infection, and in an environment with staphylococcus aureus biofilm infection, baohuogan I can effectively inhibit the formation of staphylococcus aureus biofilm and can be combined with other antibiotics to effectively remove the formed staphylococcus aureus biofilm, so that the adverse prognosis problems of repeated chronic infection, delayed healing and the like caused by staphylococcus aureus biofilm infection can be reduced.
Drawings
FIG. 1 is a graph of the growth of Staphylococcus aureus at various concentrations of baohuogan I according to an embodiment of the present invention; wherein a) is SA113 Staphylococcus aureus, b) is YUSA139 Staphylococcus aureus.
FIG. 2 shows the OD of Staphylococcus aureus biofilm after crystal violet staining under different concentrations of baohuogan I according to the example of the present invention570Analyzing the detected value; wherein a) is SA113 Staphylococcus aureus, b) is YUSA139 Staphylococcus aureus. Comparison with the non-dosed group: p<0.001(Student’s t test)。
FIG. 3 is an original image of Staphylococcus aureus biofilm crystal violet staining under different concentrations of baohuogan I according to an embodiment of the present invention; wherein a) is SA113 Staphylococcus aureus, b) is YUSA139 Staphylococcus aureus.
FIG. 4 is an analysis graph of biofilms formed by the removal of 3 strains of MSSA by combining baohuogan I and daptomycin at different concentrations in the examples of the present invention. Comparison with the non-dosed group:*,P<0.05；**,P<0.01；***,P<0.001(Student’s ttest)。
FIG. 5 is a block diagram of an embodiment of the present inventionFIG. 3 shows the analysis of biofilm formed by MRSA eliminated by combining baohuogan I and daptomycin at the same concentration. Comparison with the non-dosed group:*,P<0.05；**,P<0.01；***,P<0.001(Student’s ttest)。
Detailed Description
Preferred embodiments of the present invention are described in further detail below.
Example 1
Experiments that baohuogan I effectively inhibited the formation of staphylococcus aureus biofilms.
Selecting SA113 standard strain with positive biofilm formation and a clinical strain YUSA139, and culturing the strain in TSB culture medium at 37 deg.C and 220rpm/min overnight for 10-12 h. Diluting bacterial liquid 1:200 with TSBG culture medium (TSB culture medium + 0.5% glucose) containing Baohuogan I with different concentrations, adding 96-well plate (Costar3599) into each 200ul of bacterial liquid, setting 3 multiple wells for each bacterial liquid, standing and culturing at 37 ℃ for 24h to form mature biofilm, eluting with PBS 3 times (200 ul/well/time), drying at room temperature, adding methanol for fixing for 15min (200 ul/well), discarding methanol, drying at room temperature, adding 0.5% crystal violet dye solution into each well, dyeing at room temperature for 10min, gently eluting the crystal violet dye solution under clear water until running water is colorless, drying at room temperature, and reading OD on an enzyme labeling instrument570The value is obtained.
The above experimental procedure was independently repeated 3 times, and the data were expressed as mean ± standard deviation (mean ± SD). As shown in fig. 1 to 3, it was found that baohuogan I can significantly inhibit the formation of SA113 biofilm at concentrations greater than or equal to 12.5 μ M, while baohuogan I can significantly inhibit the formation of SA113 biofilm at concentrations greater than or equal to 6.25 μ M for YUSA 139. Meanwhile, the growth curve of staphylococcus aureus strain planktonic bacteria under the action of baohuogan I is measured by a growth curve instrument (Bioscreen C, Turku, Finland), and an SA113 standard strain and a clinical strain YUSA139 are cultured in a TSB culture medium at 37 ℃ and 220rpm/min overnight for 10-12h with bacteria shaking. Bacterial liquid 1:200 is diluted by TSBG culture medium (TSB culture medium + 0.5% glucose) containing baohuogan I with different concentrations, 200ul of each hole is added into a matched hole plate (Bioscreen C, Turku, Finland), 3 multiple holes are arranged on each strain, and the baohuogan I does not influence the growth of planktonic bacteria of the two strains within 100 mu M.
Example 2
The baohuogan I can effectively inhibit the formation of 108 clinically separated staphylococcus aureus biofilms
108 clinical separated staphylococcus aureus strains are selected and cultured in a TSB culture medium at 37 ℃ and 220rpm/min overnight for 10-12 h. Diluting bacterial liquid 1:200 with TSBG culture medium (TSB culture medium + 0.5% glucose) containing 25 μ M baohuogan I, adding 96-well plate (Costar3599) into 200ul of each well, setting 3 multiple wells for each bacterial strain, standing and culturing at 37 ℃ for 24h to form mature biofilm, eluting with PBS 3 times (200 ul/well/time), drying at room temperature, adding methanol for fixing for 15min (200 ul/well), discarding methanol, drying at room temperature, adding 0.5% crystal violet dye solution into each well, dyeing at room temperature for 10min, gently eluting the crystal violet dye solution under clear water until running water is colorless, drying at room temperature, reading OD on an enzyme labeling instrument570The value is obtained. As shown in Table 1, it was found that BAOGUOGAN I can inhibit the formation of Staphylococcus aureus biofilm by 96.3% at 25. mu.M.
TABLE 1 minimum inhibitory concentration of baohuogan I (25. mu.M) against Staphylococcus aureus
Note: slightly inhibiting biofilm formation by 10% or more and less than 40%; moderate inhibition, wherein the inhibition is more than or equal to 40 percent and the biofilm formation is less than 70 percent; obviously inhibits the biofilm formation by more than or equal to 70 percent.
Example 3
The baohuogan I and daptomycin are combined to effectively remove the biofilm formed by staphylococcus aureus.
Methicillin-sensitive Staphylococcus aureus (MSSA) SA113, YUSA86, CHS134 strain, and methicillin-resistant Staphylococcus aureus (MRSA) YUSA139, YUSA142, and YUSA145 strain were cultured in TSB medium at 37 deg.C and 220rpm/min overnight for 10-12 h. Bacterial suspension 1:200 was diluted with TSBG medium (TSB medium + 0.5% glucose), 96-well plates (Costar3599) were added to 200. mu.l/well, 3 wells were set for each strain, and a mature biofilm was formed by static culture at 37 ℃ for 24 hours. Discarding supernatant, and sterilizingEluting with saline water for 3 times (200 ul/hole/time), adding fresh TSBG culture medium (containing different concentrations of BAOGUOGAN I and daptomycin combination), standing for 48h (replacing 1 new culture medium every 24 h), discarding supernatant, eluting with PBS for 3 times (200 ul/hole/time), drying at room temperature, adding methanol for fixing for 15min (200 ul/hole), discarding methanol, drying at room temperature, adding 0.5% crystal violet dye solution into each hole for 100ul, dyeing at room temperature for 10min, gently eluting the crystal violet dye solution with clear water until colorless running water, drying at room temperature, and reading OD on microplate reader570The value is obtained. The above experimental procedure was independently repeated 3 times, and the data were expressed as mean ± standard deviation (mean ± SD). As shown in FIGS. 4 and 5, when the concentrations of baohuogan I are 1.56. mu.M, 12.5. mu.M and 50. mu.M, the baohuogan I can be used together with daptomycin to obviously remove the biofilm formed by the strain YUSA139 of Staphylococcus aureus; when the concentration of baohuogan I is 50 mu M, the baohuogan I can be combined with daptomycin to obviously remove the biofilm formed by staphylococcus aureus YUSA86 and YUSA145 strains.
The embodiment of the invention discloses an application of baohuogan I in preparation of a medicine for resisting staphylococcus aureus infection, wherein the medicine for resisting staphylococcus aureus infection comprises baohuogan I which is used for improving the pharmaceutical anti-biofilm capacity of the medicine. Preferably, in the anti-staphylococcus aureus infection medicine, the concentration of baohuogan I is less than or equal to 100 mu M. The medicine can be in the form of spray, oral granule, oral tablet or injection. Preferably, the concentration of baohuogan I is not less than 12.5 mu M. Preferably, the concentration of baohuogan I is not less than 25 μ M.
Preferably, the anti-staphylococcus aureus drug comprises daptomycin. The baohuogan I and daptomycin are combined, so that the effect of resisting staphylococcus aureus infection is better.
The embodiment of the invention also discloses an application of baohuogan I in preparing a coating for the surface of a medical device, wherein the coating for the surface of the medical device comprises baohuogan I. The baohuogan I is used for inhibiting the formation of a staphylococcus aureus biofilm and the adhesion of staphylococcus aureus. Preferably, in the coating for the surface of the medical device, the concentration of baohuogan I is less than or equal to 100 mu M. Preferably, the concentration of baohuogan I is 6.25 to 100 μ M. Preferably, the concentration of baohuogan I is not less than 12.5 mu M. Preferably, the concentration of baohuogan I is not less than 25 μ M.
The embodiment of the invention also discloses a coating for the surface of a medical apparatus, which comprises baohuogan I. Preferably, the concentration of baohuogan I is 6.25 to 100 μ M. Preferably, the concentration of baohuogan I is not less than 12.5 mu M. Preferably, the concentration of baohuogan I is not less than 25 μ M. Preferably, the concentration of baohuogan I is less than or equal to 100 mu M.
The embodiment of the invention also discloses application of the baohuogan I in preparation of a disinfectant for resisting staphylococcus aureus, wherein the disinfectant comprises the baohuogan I which is used for inhibiting the formation of a biofilm of the staphylococcus aureus and the adhesion of the staphylococcus aureus.
The embodiment of the invention also discloses a disinfectant for resisting staphylococcus aureus, which comprises baohuogan I. Preferably, the concentration of baohuogan I is 6.25 to 100 μ M. Preferably, the concentration of baohuogan I is not less than 12.5 mu M. Preferably, the concentration of baohuogan I is not less than 25 μ M. Preferably, the concentration of baohuogan I is less than or equal to 100 mu M.
The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and it is not intended that the invention be limited to these specific details. For those skilled in the art to which the invention pertains, several simple deductions or substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.
Claims (10)
1. The application of baohuogan I in resisting staphylococcus aureus biofilm infection is characterized in that: baohuogan I is used for resisting staphylococcus aureus biofilm infection.
2. Use of baohuogan I according to claim 1 for the treatment of staphylococcus aureus biofilm infection, characterized in that: the concentration of the baohuogan I is 6.25 mu M-100 mu M, and preferably, the concentration of the baohuogan I is not less than 12.5 mu M; preferably, the concentration of baohuogan I is not less than 25 μ M.
3. The application of baohuogan I in preparing the medicines for resisting staphylococcus aureus infection is characterized in that: the medicine for resisting staphylococcus aureus infection comprises baohuogan I.
4. The application of baohuogan I in preparing a medicine for resisting staphylococcus aureus infection according to claim 3 is characterized in that: in the medicine for resisting staphylococcus aureus infection, the concentration of baohuogan I is less than or equal to 100 mu M.
5. The application of baohuogan I in preparing the medicines for resisting staphylococcus aureus infection according to claim 4 is characterized in that: the anti-staphylococcus aureus drug comprises daptomycin.
6. The application of baohuogan I in preparing the coating for the surface of medical instruments is characterized in that: the coating for the surface of the medical instrument comprises baohuogan I.
7. The use of baohuogan I according to claim 6 in the preparation of coatings for medical device surfaces, characterized in that: in the coating for the surface of the medical instrument, the concentration of baohuogan I is less than or equal to 100 MuM.
8. A coating for a medical device surface, characterized by: it comprises baohuogan I.
9. The application of baohuogan I in preparing a disinfectant for resisting staphylococcus aureus is characterized in that: the disinfectant for resisting staphylococcus aureus comprises baohuogan I.
10. A disinfectant against staphylococcus aureus bacteria, comprising: it comprises baohuogan I.
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| CN1969871A (en) * | 2005-11-24 | 2007-05-30 | 北京奇源益德药物研究所 | Antivirus compound formulation, preparation process, quality control method and use thereof |
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| CN1969871A (en) * | 2005-11-24 | 2007-05-30 | 北京奇源益德药物研究所 | Antivirus compound formulation, preparation process, quality control method and use thereof |
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