KR101783267B1 - A composition comprising alismatis rhizoma extract for preventing, improving or treating periodontal disease - Google Patents
A composition comprising alismatis rhizoma extract for preventing, improving or treating periodontal disease Download PDFInfo
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- KR101783267B1 KR101783267B1 KR1020150165951A KR20150165951A KR101783267B1 KR 101783267 B1 KR101783267 B1 KR 101783267B1 KR 1020150165951 A KR1020150165951 A KR 1020150165951A KR 20150165951 A KR20150165951 A KR 20150165951A KR 101783267 B1 KR101783267 B1 KR 101783267B1
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- periodontal disease
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/312—Foods, ingredients or supplements having a functional effect on health having an effect on dental health
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2300/00—Processes
- A23V2300/14—Extraction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
Abstract
The present invention relates to a composition for preventing, ameliorating or treating periodontal disease containing an extract of Alismatis rhizoma as an active ingredient.
The composition according to the present invention has an efficacy that corresponds to a minimum inhibitory concentration (MIC) value of 156 to 2500 占 퐂 / ml and a minimum bactericidal concentration (MBC) value of 625 to 5000 占 퐂 / ml for pathogenic bacteria of periodontal disease, respectively .
The present invention also relates to a composition for preventing, ameliorating or treating periodontal disease containing a mixture of a taxa extract and a conventional antimicrobial substance as an active ingredient.
The composition according to the present invention has an effect of causing a synergistic effect by containing a mixture of a taxa extract and a conventional antimicrobial substance as an active ingredient, treating a mixture of a 1/2 MIC value of a taxa extract and a conventional antimicrobial substance, After 10 hours, the growth of periodontal disease pathogens is completely killed.
Description
The present invention relates to a composition for preventing, ameliorating or treating periodontal disease containing an extract of Alismatis rhizoma as an active ingredient.
Specifically, the extract has a minimum inhibitory concentration (MIC) value of 156-2500 占 퐂 / ml and a minimum bactericidal concentration (MBC) of 625 ~ 5000 占 퐂 / ml for pathogenic bacteria of periodontal disease .
The present invention relates to a composition for preventing, ameliorating or treating periodontal disease containing a mixture of a taxa extract and a conventional antibacterial substance as an active ingredient.
Specifically, it has an effect of causing a synergistic effect by containing a mixture of a taxa extract and a conventional antimicrobial substance as an active ingredient, treating a mixture of the MIC value of the taxa extract with a conventional antimicrobial substance and treating the mixture for 12 to 18 hours Later, the growth of periodontal disease pathogens is completely dead.
The plaque is a part of the normal oral microorganism gun gathered on the tooth surface. Plaque develops its own homogeneous distribution and structure balance (homeostasis) without causing any problems to the human body, unless there is a special environmental change, and the number of bacteria increases as it matures.
However, changes in the oral or plaque environment, changes in the prefrontal cortex, or the like can lead to a significant increase in the number of specific bacterium species in the teeth or an increased chance of bacterial poisoning, leading to opportunistic infections leading to teeth and periodontal tissues Which causes periodontal disease while acting on the teeth.
Periodontal disease is an inflammatory disease that occurs in the periodontal tissue, the supporting tissue of the teeth surrounding the tooth. Periodontal disease is often referred to as taste, depending on the degree of disease gingivitis (gingivitis) and periodontitis (periodontitis) is divided into.
It is a relatively light and fast-acting periodontal disease called gingivitis that is limited to gingiva or soft tissue, and it is called periodontitis when the inflammation progresses to the gums and the gums.
There is a V-shaped gap between the gum (gums) and the teeth. Periodontal disease is caused by bacteria attacking the lower part of the gum line of this sulcus and damaging the periodontal ligament and adjacent tissues. As the inflammation progresses and more tissue is damaged, the groove develops into a periodontal pocket, and the more the periodontal disease becomes, the deeper the depth of the periodontal pocket becomes. Periodontal disease is a periodontal disease that causes inflammation and bone loss in the periodontal ligament.
The narrow space between the teeth and gums (gingival sulcus, gingival crevice) is an anaerobic environment with relatively less oxygen than the exposed area. This space becomes deeper if there is periodontal disease and the anaerobic state becomes worse. This new supply of nutrients and growth factors, as well as the anaerobic environment, is dominated by rod-shaped Gram negative anaerobic bacteria, ie, oxygen-free environment-favored bacteria.
Periodontal disease associated with periodontal disease includes Actinobacillus actinomycetemcomitans , Porphyromonas gingivalis , Tannerella forsythia , Treponema spp., Treponema spp. denticola ), But are not limited to , Fusobacterium nucleatum, Peptostreptococcus micros, Streptococcus intermedius , Prevotella intermedia , intermedia ) and And Prevotella nigrescens , among which Porphyromonas ( Porphyromonas) gingivalis , together with Tannerella forsythia and Treponema denticola , are called red complexes and are classified as the most dangerous group of periodontal disease bacteria.
These red complex bacteria have a much greater proteolytic ability than other oral bacteria, and this ability can further exacerbate tissue destruction. However, more severe tissue destruction occurs during the course of these bacterial and tissue cells, fungi and immune cells. For example, various proteolytic enzymes secreted by these bacteria, or matrix metalloproteinases (MMPs), which are tissue-destroying enzymes secreted from tissue cells or phagocytic cells, ). ≪ / RTI > Activated MMP accelerates the destruction of periodontal tissue, and the various inflammation inducers secreted by human cells in response to bacteria in the red complex further intensify the destruction of periodontal tissue.
Periodontal disease is a chronic inflammatory disease. Therefore, periodontal disease can affect the systemic disease that chronic inflammation is a fundamental mechanism of onset. When inflammatory substances (cytokines, etc.) are secreted from inflammatory sites, they may act on other tissues as a primary cause, or inflammation-inducing bacteria may access other tissues and cause systemic diseases. For this reason, periodontal disease can be a cause of heart disease (especially myocardial infarction) associated with atherosclerosis, and the mode of the disease may be the cause of ischemic stroke, basically similar to myocardial infarction.
Methods for the improvement or treatment of periodontal disease include, but are not limited to, the use of a toothpaste called chlorhexidine, the introduction of special medicines between the gums and teeth, and the use of antibiotics to eradicate specific germs in the gums .
However, these methods are complicated, costly and time consuming. In addition, when the oral bacteria are eradicated using antibiotics, the balance of the bacteria in the oral cavity may collapse, resulting in decreased oral immunity, dry mouth and other diseases, and resistance of oral bacteria to antibiotics may occur .
Therefore, it is necessary to develop alternative substances that can prevent, improve or treat oral diseases such as periodontal diseases.
Accordingly, the inventors of the present invention have found a composition that can prevent, ameliorate or treat periodontal disease without using side effects due to the use of a substance derived from a natural source.
The composition according to the present invention is characterized by containing a taxa extract as an active ingredient and has a synergistic effect when mixed with an existing antimicrobial substance.
A brief description of Alismatis rhizoma used in the present invention is as follows.
Alismatis rhizoma is a species of Alisma orientale Juzepzuk) as the root of the root of the root is removed. It is spherical, elliptical or oval, 2 to 7 cm in length, and 2 to 6 cm in diameter. Outer is yellowish white to light yellowish brown, body is light and quality is solid. The cut surface is yellowish white and powdery, and has many fine holes.
Taxa is a diuretic, lowers blood sugar, has antimicrobial activity and is used for cystitis, urethritis, nephritis and hypertension.
Alasol, alismol, alismoxide and monoacetate compounds are included in the taxa. Others include large amounts of starch and protein, beta-sitosterol, ledithin and choline.
The present applicant has completed the present invention by providing a composition for measuring the antibacterial activity against the periodontal disease bacteria of the taxa extract and for preventing, improving or treating periodontal disease containing the same.
The prior art related to the present invention is described in Korean Patent Registration No. 10-1025886 (Prior Art 1), which discloses a composition for inhibiting tooth decay, which contains a taxa extract.
Although the composition containing the extract of Phytophthora exudata has been described in the prior art document 1, only the antimicrobial activity against Streptococcus mutans , which is a causative organism of caries, has been confirmed.
Oral diseases can be divided into dental caries (tooth decay) and periodontal disease (taste). Dental caries and periodontal disease are different diseases, and the causative bacteria are different.
It is known that the acid produced by the decomposition of the dental caries bacteria causes corrosion of the enamel on the tooth surface. It is known that the gram-positive bacterium Streptococci is the strongest dental caries inducer. Streptococcus species include Streptococcus mutans , Streptococcus sanguinis , Streptococcus sobrinus , Streptococcus ratti , Streptococcus criceti , Streptococcus spp ., Streptococcus spp ., Streptococcus spp . Streptococcus anginosus , Streptococcus gordonii , and the like. In addition, the lactic acid bacteria belonging to the genus bacillus (Lactobacillus), actinomycetes (Actinomyces) known as dental caries pathogen.
As described above, the periodontal disease Tino Tino evil and evil is a causative agent Bacillus My setem Komi Tansu (Actinobacillus actinomycetemcomitans), Fort fatigue Monastir long lease Bangui (Porphyromonas gingivalis , Tannerella forsythia , Treponema denticola , Fusobacterium < RTI ID = 0.0 > nucleatum , Peptostreptococcus micros, Streptococcus intermedius , Prevotella intermedia , and the like. And Prevotella nigrescens .
Therefore, the antimicrobial activity against the pathogenic microorganisms related to periodontal disease of the present invention can not be confirmed from the preceding document 1 which confirmed only the antimicrobial activity against Streptococcus mutans , a causative organism of dental caries.
It is an object of the present invention to provide a composition for preventing, ameliorating or treating periodontal disease containing a naturally occurring substance as an active ingredient.
It is also an object of the present invention to provide a composition for preventing, ameliorating or treating periodontal disease containing Alismatis rhizoma extract as an active ingredient.
It is also an object of the present invention to provide a method for the treatment of periodontal disease in which the extract has a minimum inhibitory concentration (MIC) value of 156 to 2500 占 퐂 / ml and a minimum bactericidal concentration (MBC) value of 625 to 5000 占 퐂 / Thereby preventing, ameliorating or treating the disease.
It is also an object of the present invention to provide a composition for preventing, ameliorating or treating periodontal disease containing a mixture of a taxa extract and a conventional antibacterial substance as an active ingredient.
It is also an object of the present invention to provide a composition for preventing, ameliorating or treating periodontal disease which has an effect of causing a synergistic effect by containing a mixture of a taxa extract and a conventional antibacterial substance as an active ingredient.
It is also an object of the present invention to provide a method of preventing, ameliorating, or treating periodontal disease, which is effective in completely eliminating the growth of periodontal disease pathogens after 12 to 18 hours of treatment with a mixture of a conventional antimicrobial compound with a 1/2 MIC value Compositions.
The present invention aims at solving the technical problem by providing a composition for preventing, ameliorating or treating periodontal disease containing an extract of Alismatis rhizoma as an active ingredient.
The present invention also relates to a method for the treatment of periodontal disease, wherein the extract has a minimum inhibitory concentration (MIC) value of 156 to 2500 占 퐂 / ml and a minimum bactericidal concentration (MBC) value of 625 to 5000 占 퐂 / Preventing, ameliorating, or treating a disease or condition.
In addition, the present invention aims to solve the technical problem by providing a composition for preventing, improving or treating periodontal disease containing a mixture of a taxa extract and a conventional antimicrobial substance as an active ingredient.
In addition, the present invention aims to solve the technical problem by providing a composition for preventing, ameliorating or treating periodontal disease which has an effect of causing a synergistic effect by containing a mixture of a taxa extract and a conventional antibacterial substance as an active ingredient.
The present invention also provides a composition for preventing, ameliorating, or treating periodontal disease, which is effective for treating a mixture of a 1/2 MIC value of a taxa extract and a conventional antimicrobial substance and completely eliminating the growth of periodontal disease pathogens after 12 to 18 hours To solve technical problems.
The composition according to the present invention may be provided as a composition for preventing, ameliorating or treating periodontal disease, and each composition may be provided as a health functional food using a foodstuff acceptable processing agent. The health functional food using the composition according to the present invention may be provided in various forms such as a functional beverage, a health supplement, tea, confectionery, and the like.
In order to use the present invention as a pharmaceutical composition, it may be prepared by a method well known in the pharmaceutical field, and may be mixed with the carrier itself, or a pharmaceutically acceptable carrier, excipient, diluent or the like to prepare a powder, a granule, a tablet, Injections, and the like. They may also be administered orally or parenterally.
The effective dose of the composition according to the present invention can be appropriately selected depending on the degree of absorption of the active ingredient in the body, the rate of water activation and excretion, the age, sex and condition of the patient, severity of the disease to be treated,
Formulations such as pills, granules, beverages, tablets, capsules and the like may be prepared using the composition according to the present invention, and in this case, a treatment agent may be added to prepare each formulation.
The extract of Alismatis rhizoma used as an active ingredient in the composition according to the present invention has an effect of preventing, ameliorating or treating periodontal disease.
In the composition according to the present invention, the extract of the present invention has a minimum inhibitory concentration (MIC) value of 156 to 2500 占 퐂 / ml and a minimum bactericidal concentration (MBC) value of 625 to 5000 占 퐂 / ml for pathogenic bacteria of periodontal disease There is a corresponding effect.
The composition according to the present invention has the effect of causing a synergistic effect on the periodontal disease by the taxa extract and the existing antimicrobial substance.
The composition according to the present invention is effective to treat the mixture of the 1/2 MIC value of the extract with the conventional antimicrobial substance and to completely kill the growth of periodontal disease pathogens after 12 to 18 hours.
FIG. 1 is a graph showing the time course of treatment with actinomycetes actinomycetemcomitans , a mixture of taxa extract and ampicillin, a mixture of taxa extract and gentamicin, Of the present invention.
FIG. 2 is a graph showing the time-dependent changes in the time-course of treatment with the taxa extract-treated group treated with F. nucleatum , a mixture of taxa extract and ampicillin, and the group treated with taxa extract and gentamicin This is a graph showing the antibacterial effects.
FIG. 3 is a graph showing the time-dependent antimicrobial activity of the group treated with the taxa extract treated with periodontal bacteria, P. gingivalis , the mixture of the mixture of taxa extract and ampicillin, and the mixture of taxa extract and gentamicin Fig.
The terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms and the inventor may properly define the concept of the term in order to best describe its invention It should be construed as meaning and concept consistent with the technical idea of the present invention.
Therefore, the experimental examples and the reference examples described in the present specification are merely the most preferred embodiments of the present invention and are not intended to represent all of the technical ideas of the present invention. Therefore, various equivalents and variations Examples should be understood.
Example
One.
Tarsa
Preparation of extract
Alismataceae (Aliamatis rhizoma, stem pits of fine roots and Plantain Alismataceae (Alisma orientale Juzepzuk) removing the jupi) were purchased from Cooperative Herb JEONBUK (herbal medicine cooperative association of jeonbuk, korea)).
The extraction method for extracting the taxa extract may be various extraction methods such as a solvent extraction method, a steam distillation method, a carbon dioxide supercritical extraction method, a microwave processing extraction method and a percolation extraction method, and a solvent extraction method may be preferably used.
When the solvent is extracted, the extraction temperature, the extraction time, the amount of the solvent and the treatment method of the residual component are treated differently depending on the type of the extraction solvent. Extraction solvents. Various solvents may also be used.
Extractable solvents include water, ethanol, methanol, fatty oil, glycerin, propylene glycol, ether, chloroform, petroleum ether Hexane, benzene, methylene chloride, ethyl acetate, acetone, butanol, and isopropanol. Preferably, ethanol is used. .
The solvent extraction can be carried out at a temperature of 60 to 100 ° C for 2 to 6 hours, preferably at a temperature of 80 ° C for 4 hours.
Further, the extraction, filtration, freeze-drying, and the like may be performed one or more times according to the design conditions, and may be dissolved in the solvent after performing the above-mentioned process.
The solvent can be concentrated by removing the solvent under vacuum using a rotary evaporator, preferably at 40 캜, and dissolved in 10% DMSO (dimethyl sulfoxide).
Experimental Example
1. Minimum inhibitory concentration (
MIC
) And the analysis of the minimum bactericidal concentration (MBC)
1-1. Preparation for experiment
The periodontal disease bacteria used in this experiment were Actinobacillus actinomycetemcomitans (ATCC 43717), Fusobacerium nucleatum (ATCC 10953) and Poppyromonas gingivalis ( Porphyromonas gingivalis ) (ATCC 33277).
ATCC stands for American type culture collection.
The preparation of the experiment is as follows.
(Sigma, St. Louis, Mo., USA) and menadione 1 ㎍ / ㎖ (Sigma) were inoculated with BHI.
1-2. Experimental course
Liquid dilution test (broth dilution test) is a test to determine the activity of a specific antimicrobial agent against isolated bacteria. Liquid dilution culture methods include MIC (Minimum inhibitory concentration) and MBC (Minimun bactericidal contentration) measurements.
MIC is the minimum amount of antimicrobial needed to inhibit or bacterium growth and MBC is the minimum amount of antimicrobial needed to kill bacteria. MBC is defined as the minimum concentration of antibiotics that reduces the amount of pathogens by 99.9% (ie, from 10 5 to 6 / mL to ≤ 10 2 to 3 / mL).
Most of the antimicrobial antimicrobials decrease the bacteria by more than 99.9% at the concentration of MIC, so the majority of MIC and MBC are the same. In general, it is reasonable to predict the sterilization by MIC alone. However, if it is necessary to use antibiotics with high sterilization power to remove the causative organisms due to defects before host defense such as meningitis, endocarditis, neutropenia, MBC It is used as an indicator.
First, MIC was repeated three times in order to measure the antibacterial activity of the extract from the taxa. The MIC analysis method is as follows.
(1) Bacteria cultured in the procedure of Experimental Example 1-1 were cultured under anaerobic conditions at 37 ° C. Among the cultured bacteria, aerobic bacteria were cultured for 24 hours, tuber - anaerobic bacteria were cultivated for 18 hours, and organisms were cultivated for 1-2 days. Here, tuber-anaerobic bacteria are bacteria that can develop regardless of the presence or absence of oxygen. Organic anaerobes are bacteria that can not develop if oxygen is present.
② Taxa extract, ampicilin, and gentamicin were diluted 3 times. Ampicillin and gentamicin were tested as positive controls.
③ Liquid medium was dispensed into a test tube, and the three times diluted extracts of taxa, ampicillin, and gentamicin were treated by concentration.
④ The tube was treated with the strain of the bacteria cultured by the procedure of ①.
⑤ After incubating the tubes at 37 ℃ for 18 ~ 48 hours, the tubes were observed with the naked eye and the lowest concentration of tubes was determined by MIC. MIC 50 refers to a 50% minimum inhibitory concentration of bacterial growth, and MIC 90 refers to a 90% minimum inhibitory concentration of bacterial growth.
Next, MBC was performed to measure the antimicrobial activity of the extract of the taxa. The MBC analysis method is as follows.
① For MIC analysis, the medium cultured in the above procedure was visually observed, and the medium in which the growth of the strain was completely inhibited was selected and spread on a 96-well plate.
② The number of live bacteria was measured in the medium.
1-3. Experiment result
[Table 1] shows the results of MIC and MBC of the group treated with the taxa extract, the group treated with ampicillin, and the group treated with gentamicin.
ATCC 43717
ATCC 10953
ATCC 10558
As a result of the measurement, the actinomycetemcomitans , actinomycetemcomitans , periodontal bacteria, F. nucleatum and P. gingivalis , The MIC values of the extract-treated group were all 2500 占 퐂 / ml.
The MIC value of the positive control group, Ampicillin, was at least 32 ㎍ / ㎖ at a minimum of 0.25 ㎍ / ㎖, and the MIC value of the other positive control group, gentamicin, was at least 8 ㎍ / ㎖ and a maximum of 16 ㎍ / ㎖.
The lower the MIC value, the higher the susceptibility to antibiotics. The higher susceptibility to antibiotics means higher antimicrobial activity.
Therefore, the order of antimicrobial activity based on the MIC values is listed in order of ampicillin, gentamicin, and taxa extract.
The lower the value of MBC, the lower the MIC value, the higher the susceptibility to antibiotics and the higher the antibacterial activity against bacteria.
The MBC value of the extract was 5000 ㎍ / ㎖, and the positive control group showed the highest antimicrobial activity among the three experimental groups with MBC value of at least 0.5 ㎍ / ㎖ and maximum 64 ㎍ / ㎖, MBC value was at least 8 ㎍ / ㎖ and maximum 32 ㎍ / ㎖.
Therefore, the order of antimicrobial activity based on MBC values is listed in order of ampicillin, gentamicin, and taxa extract.
Experimental Example
2.
Checker board
Dilution (Checker-board dilution) experiment
2-1. Preparation for experiment
The same procedure as in Experimental Example 1-1 was carried out.
2-2. Experimental course
Synergistic action is a phenomenon in which the effect of two different drugs at the same time within a dose is stronger than the effect of each drug alone when administered alone, ) Action is the same as or similar to the effect of two different drugs at the same dose in the same dose, plus the effect of each drug alone.
Fractional inhibitory concentration index (FICI) is the sum of the FIC (Fractional inhibitory concentration) values of the most effective combinations and is most commonly used to interpret the results of antibiotic combination studies. When the FICI is less than 0.5, the synergistic effect is obtained. When the FICI is 2.0 or more, the antagonistic action is judged.
The following equation (1) is a calculation method of FIC.
Here, (A) refers to the MIC when the drug to be used in combination with the drug A is used together, and MIC A refers to the MIC when the drug A is used alone.
Equation (2) below is a calculation method of FICI.
FBCI (fractional bactericidal concentration index) is the sum of the values of FBC (fractional bactericidal concentration) in the most effective combination. The calculation method is the same as the calculation method of FICI.
Experiments that measured the synergistic effect of mixing ampicillin or gentamicin with the extract of the taxa were carried out using Checker-Board Dilution.
(1) In the same manner as in Experimental Example 1, periodontal disease bacteria were inoculated into the medium and cultured.
② Each extract was treated with either ampicillin or gentamicin to prepare each mixture.
③ The prepared mixture was serially diluted.
(4) Treat 96 well microplates with Mueller-Hinton liquid medium supplemented with cations and then treat the series diluted mixture.
⑤ Sequence The diluted mixture is treated with the culture solution cultured on the treated medium.
⑥ The medium treated with the strain was incubated at 37 ° C for 24 to 48 hours.
⑦ The MIC and MBC analyzes were performed in the same manner as in Experimental Examples 1 and 2.
2-3. Experiment result
Table 2 shows the results of the checkerboard dilution experiment when the extract of the taxa alone was treated and when the mixture of the taxa extract and the ampicillin was treated.
ATCC 43717
Ampicillin
32/64
16/32
Appositional
ATCC 10953
Ampicillin
1/4
0.25 / 1
0.25 / 0.25
Appositional
ATCC 10558
Ampicillin
0.25 / 0.5
0.125 / 0.25
Appositional
As a result of the measurement, the MIC of the extract obtained by treating the extract alone was 2500 g / ml, and the MBC was 5000 g / ml.
The MIC of ampicillin alone was at least 32 ㎍ / ㎖ at a minimum of 0.25 ㎍ / ㎖, and the maximum MBC of 64 ㎍ / ㎖ at a minimum of 0.5 ㎍ / ㎖.
The MIC of the extracts of Taxa extract and amphycillin was 1250 ㎍ / ㎖ at a minimum of 625 ㎍ / ㎖, and 2500 ㎍ / ㎖ at a minimum of 1250 ㎍ / ㎖.
MIC of ampicillin when mixed with amoxicillin extract was at least 0.1 μg / ㎖ and maximum 16 μg / ㎖, and MBC was at least 0.2 ㎍ / ㎖ and maximum 32 ㎍ / ㎖.
The FIC of the extracts was at least 0.5 ㎍ / ㎖ at a minimum of 0.25 ㎍ / ㎖ and the maximum was 0.5 ㎍ / ㎖ at a minimum of 0.25 ㎍ / ㎖.
The FIC of ampicillin ranged from a minimum of 0.25 ㎍ / ㎖ to a maximum of 0.5 ㎍ / ㎖, and the FBC also ranged from a minimum of 0.25 ㎍ / ㎖ to a maximum of 0.5 ㎍ / ㎖.
The FICI of phytase extract and ampicillin was 0.75 ㎍ / ㎖. In addition, the FBCI of phytase extract and ampicillin was 0.75 ㎍ / ㎖.
The FICI or FBCI most commonly used in the analysis of the antibiotic combination effects is synergistic if the value is less than 0.5, antagonistic if more than 2.0,
As a result of the experiment, it was confirmed that FICI and FBCI were 0.75 ㎍ / ㎖ when the extract was mixed with ampicillin. Further, it was confirmed that it was very close to the synergism reference value (0.5) in the adsorption action range (more than 0.5 and less than 2.0).
That is, the MIC and MBC when mixed with the taxa extract and the ampicillin were lower than the MIC and MBC when treated with the extract of the taxa and the ampicillin, respectively.
[Table 3] shows the results of a checkerboard dilution experiment when the extract of the taxa alone was treated and the mixture of the taxa extract and the gentamicin was treated.
ATCC 43717
Gentamicin
8/8
2/2
0.25 / 0.25
Synergistic
ATCC 10953
Gentamicin
16/32
2/8
0.125 / 0.25
Synergistic
ATCC 10558
Gentamicin
16/32
4/4
0.25 / 0.125
Synergistic
As a result of the measurement, the MIC of the extract obtained by treating the extract alone was 2500 g / ml, and the MBC was 5000 g / ml.
The MIC of gentamicin alone was at least 16 ㎍ / ㎖ at a minimum of 8 ㎍ / ㎖ and the maximum was 32 ㎍ / ㎖ at a minimum of 8 ㎍ / ㎖.
The MIC of the extracts of Taxa extract was 625 ㎍ / ㎖ at the minimum of 156 ㎍ / ㎖ and 625 ㎍ / ㎖ of MBC when the extract was mixed with gentamycin.
The MIC of gentamicin when mixed with Gentamicin was 2 ㎍ / ㎖ and 4 ㎍ / ㎖ at the maximum, and MBC was at least 2 ㎍ / ㎖ and 8 ㎍ / ㎖ at the maximum.
The FIC of the extract was at least 0.063 ㎍ / ㎖ at the maximum of 0.25 ㎍ / ㎖ and FBC was at 0.125 ㎍ / ㎖.
The FIC of gentamicin was a maximum of 0.25 ㎍ / ㎖ at a minimum of 0.125 ㎍ / ㎖ and a maximum of 0.25 ㎍ / ㎖ at a minimum of 0.125 ㎍ / ㎖.
FICI of phytase extract and gentamicin ranged from at least 0.313 ㎍ / ㎖ to a maximum of 0.5 ㎍ / ㎖. Therefore, it can be said that the FICI is highly synergistic when mixed with the extract of Taxa and gentamicin.
The FBCI of the taxa extract and gentamicin was at least 0.25 ㎍ / ㎖ and the maximum was 0.375 ㎍ / ㎖. All three treatment groups showed synergistic effect. Therefore, FBCI is also highly synergistic when mixed with GPT and gentamicin.
In other words, the MIC and MBC were lower when the taxa extract and gentamicin were mixed with each other than the MIC and MBC when the extract was treated with the single extract of the taxa and the gentamicin, and the synergistic effect inhibited bacterial growth and sterilization .
Experimental Example
3. Time-kill curve experiment
3-1. Preparation for experiment
The same procedure as in Experimental Example 1-1 was carried out.
3-2. Experimental course
Time-kill curves for periodontal bacteria were performed by a broth macrodilution experiment. The experimental method is as follows.
(1) Actinobacillus actinomycetemcomitans and Fusobacterium strains in BHI medium, nucleatum ) were cultured with the number of bacteria set at 6 ~ 7 × 10 6 CFU / ml. Porphyromonas gingivalis were cultured separately in hemin and menadione treated BHI medium.
② BHI medium was divided into 3 groups and treated with MIC TARGET extract, 1/2 MIC TARGET extract + 1/2 MIC ampicillin, 1/2 MIC TARGET extract + 1/2 MIC GENTAMICHIN.
③ The media were incubated at anaerobic condition at 37 ℃. At 0, 30, 1, 2, 3, 6, 9, 12, 18, 24 and 48 hours after the incubation.
(4) The number of colonies in each medium was calculated twice to obtain the average value of the number of colonies. Tino Tino Bacillus evil evil system of microstructure Komi Tansu (A. actinomycetemcomitans) and earthy Pu Te Solarium press Hinckley Atum colony count was executed in BHI medium, Monastir is long balises (P. gingivalis) to the foreskin (F. nucleatum) Colony number calculations were performed on BHI medium containing hemin and menadione.
3-3. Experiment result
FIG. 1 is a graph showing the time course of treatment with actinomycetes actinomycetemcomitans , a mixture of taxa extract and ampicillin, a mixture of taxa extract and gentamicin, Of the present invention.
FIG. 2 is a graph showing the time-dependent changes in the time-course of treatment with the taxa extract-treated group treated with F. nucleatum , a mixture of taxa extract and ampicillin, and the group treated with taxa extract and gentamicin This is a graph showing the antibacterial effects.
FIG. 3 is a graph showing the time-dependent antimicrobial activity of the group treated with the taxa extract treated with periodontal bacteria, P. gingivalis , the mixture of the mixture of taxa extract and ampicillin, and the mixture of taxa extract and gentamicin Fig.
Measurement, Ill Martino Bacillus evil Martino micro system Cormier Tansu showed the highest antibacterial activity when viewed at a time-kill curve of (A. actinomycetemcomitans) were Alismataceae extract mixture and ampicillin treatment group, followed by the Alismataceae extract and gentamicin mixture Treatment group, and taxa extract treatment group.
When the time kill curve of F. nucleatum was examined, the highest antimicrobial activity was in the mixture of taxa extract and gentamicin mixture, followed by taxa extract and ampicillin mixture treatment, The antimicrobial activity was higher in the order of treatment group.
The time kill curve of P. gingivalis showed the highest antimicrobial activity in the group treated with the taxa extract and ampicillin mixture, followed by the taxa extract treatment group, the taxa extract and the gentamicin mixture treatment The antimicrobial activity was high in the order of the group.
In summary, it can be seen that the extracts of the taxa and the existing antimicrobial substances have the effect of causing a synergistic effect on pathogenic bacteria of periodontal disease.
In other words, it can be seen that the treatment of a mixture of the antimicrobial substance with 1/2 MIC value of the extract of Phytophthora exudates completely and the growth of periodontal disease pathogen is completely weakened after 12 to 18 hours.
Claims (5)
The concentration of the extract of Alismatis rhizoma is 156 ~ 5000 / / ml,
Wherein the pathogen of the periodontal disease is Actinobacillus actinomycetemcomitans , Fusobacterium nucleatum , and Porphyromonas gingivalis .
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NITISH SURATHU 외 1명. Traditional therapies in the management of periodontal disease in India and China. Periodontology 2000, Vol. 56, 2011, pp. 14-24* |
김천종 외 3명. 八味地黃丸 및 五倍子 추출물이 뼈모유사세포와 치주인대섬유모세포의 증식, Alkaline Phosphatase의 활성 및 단백질 합성능에 미치는 影響. 대한한의학회지 제24권 제3호, 2003년 9월, pp. 35-44* |
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