JP2017001960A - Antiinflammatory agent, antioxidant and antibacterial agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an antiinflammatory agent, an antioxidant and an antibacterial agent, comprising Ocimum bacilicum, a tropical plant distributed through Brazil, Southern Europe, and China, as an active ingredient.SOLUTION: The antiinflammatory agent, antioxidant and antibacterial agent comprise one of Ocimum bacilicum whole plants, Ocimum bacilicum extracts and Ocimum bacilicum microorganism fermentation products as an active ingredient, and have rheumatoid arthritis inhibitory action, chemokine gene expression inhibitory action, DPPH radical scavenging capacity, and antibacterial action on periodontal disease bacteria.SELECTED DRAWING: None

Description

  The present invention relates to an anti-inflammatory agent, an antioxidant and an antibacterial agent comprising mannericone as an active ingredient.

  Mangelikon is a tropical plant distributed in Brazil, southern Europe, and China. It is a kind of perilla and mint, and has a fragrance in the whole grass and grows up to about 1 meter in height.

  Although this mannericon is generally used as a seasoning for meat dishes in Brazil, it has been said to have an effect of promoting insulin secretion and an effect of suppressing blood sugar levels since ancient times.

  For this reason, there are conventionally health foods, pharmaceuticals, cosmetics, and the like that contain mangelikon. As an example, a composition for health food that prevents and prevents obesity and is useful for the prevention and improvement of chronic diseases such as hypertension, hyperlipidemia, diabetes, cardiovascular disease, and heart disease, which are said to be lifestyle-related diseases. Exists. This composition for health food contains an ingredient having an appetite suppressing action, an ingredient having an inhibitory action on digestive enzyme activity, an ingredient comprising chitosan, an ingredient having dietary fiber, or an ingredient having an inhibitory action on digestive enzyme activity, mushroom The chitosan complex polysaccharide component and the mineral balance are adjusted, and the component having insulin production and insulin receptor activation is included. In addition to these components, this composition for health food contains mangelicon as a plant extract extract that does not increase blood sugar levels (Patent Documents 1 and 2).

  As another example, blood glucose level characterized by using coral fossil produced from nature, or a mineral derived from marine organisms in which calcium in limestone produced from marine organisms is mineralized by substituting magnesium in the ocean There are mineral-containing compositions with a lowering effect. In addition to these minerals, this mineral-containing composition contains at least one plant component selected from a group of plants containing mangelicon (Patent Document 3).

  As yet another example, there is a whitening cosmetic composition that is effective in preventing or treating spots, buckwheat and the like, exhibits an excellent whitening effect, and has both stability and high safety to the skin. This whitening cosmetic composition is at least selected from the group consisting of hemp, calcager, coconut, coconut, coconut, bamboo leaves, gennoshouko, banchu leaves, watafuji, chrysanthemum, walnuts, mangelikon and spicy It is assumed that one extract is contained as an active ingredient (Patent Document 4).

JP 2007-330124 A JP 2006-20606 A JP 2004-43335 A JP 7-25746 A

  Therefore, paying attention to the mangericon used in the above-mentioned health foods, pharmaceuticals, cosmetics, etc., the whole plant, extract or microbial fermented product of this mangericon has the effect of suppressing rheumatoid arthritis, suppressing the expression of chemokine genes, eliminating DPPH radicals. And the antibacterial action against periodontal disease bacteria has been found, and the anti-inflammatory agent, antioxidant and antibacterial agent of the present invention have been provided.

  The anti-inflammatory agent of the present invention has rheumatoid arthritis-inhibiting action by using any one of whole mangericon, mangericon extract and mangericon microorganism fermented product as an active ingredient.

  Furthermore, the anti-inflammatory agent of the present invention contains any of mannericon whole plant, mannellicon extract and mannericon microbial fermentation product as an active ingredient, and has a chemokine gene expression inhibitory action.

  In addition, the antioxidant of the present invention has any one of mannericon whole plant, mangericon extract and mannericon microorganism fermented product as an active ingredient, and has DPPH radical scavenging ability.

  Furthermore, the antibacterial agent of the present invention has an antibacterial action against periodontal disease bacteria by using any one of mannericon whole plant, mangericon extract and fermented mangericon microorganism as an active ingredient.

  The anti-inflammatory agent of the present invention has a rheumatoid arthritis inhibitory action and can effectively eliminate inflammation, pain, swelling, etc. of rheumatoid arthritis.

  Furthermore, the anti-inflammatory agent of the present invention has a chemokine gene expression inhibitory action, and can effectively eliminate the occurrence and amplification of inflammation such as contact dermatitis and atopic dermatitis.

  In addition, the antioxidant of the present invention has DPPH radical scavenging ability, and has the effect of preventing and improving various symptoms such as skin inflammation, dryness, wrinkles and sagging when used as a skin external preparation or cosmetic. Furthermore, when it is used for eating and drinking, it exerts effects such as promotion of suppression of active oxygen and free radicals which are cell inhibitors in the body.

  Furthermore, the antibacterial agent of the present invention has an antibacterial action against periodontal disease bacteria, Porphyromonas gingivalis and Actinomyces naeslundii, and effectively eliminates periodontal disease. As well as being able to do so, it has also been effective in preventing bad breath.

It is a graph which shows the measurement result of TNF- (alpha) value of the anti-inflammatory agent of this invention. It is a graph which shows the measurement result of the expression level of the chemokine gene of the anti-inflammatory agent of this invention. It is a graph which shows the measurement result of the scavenging ability of DPPH radical of the antioxidant of the present invention. It is a graph which shows the measurement result of the diameter of the inhibition zone with respect to periodontal disease bacteria of the antibacterial agent of the present invention.

  Hereinafter, the form for implementing the anti-inflammatory agent, antioxidant, and antibacterial agent of this invention is demonstrated in detail.

  The anti-inflammatory agent, antioxidant and antibacterial agent of the present invention contain any one of mannericon whole plant, mannericon extract and mannericon microbial fermented product as an active ingredient.

  Mangelikon in the present invention has a scientific name of Ocimum bacilicum, and as mentioned above, is a tropical plant distributed in Brazil, southern Europe, and China. It is a kind of perilla and mint, and the whole plant has fragrance and the plant height is It is a plant that grows up to about 1 meter.

  In the present invention, the whole angelica plant refers to leaves and ground stems, and may be fresh grass or dried matter.

  In the present invention, the MANGELICON extract is an extract obtained by extracting whole MANGELICON plant with cold / hot water of 4 to 60 ° C. or hot water of 60 ° C. or more, but extracted with ethanol and hydrous ethanol. It may be. Although it does not specifically limit as a property of an extract, It can be set as an extract, spray dry, a freeze-dried thing, etc.

  In the present invention, the fermented MANGERICON microorganism refers to a fermented material obtained by fermenting the entire MANGERICON plant with bacteria such as lactic acid bacteria, acetic acid bacteria and natto bacteria, fungi such as koji, yeast and the like. Furthermore, it is good also as an extract which extracted such microbial fermented material with cold / hot water, hot water, or ethanol and hydrous ethanol as mentioned above.

  The anti-inflammatory agent, antioxidant and antibacterial agent of the present invention can be used as tablets, capsules, oral liquids, drinks, etc., applied as ointments, creams, etc., toothpaste, mouth freshener, mouthwash It can be used in the mouth as a (gargle).

  In the present invention, the content of the whole mannellicon plant, the mannellicon extract and the mannellicon microbial fermentation product is 0.1 to 10% by weight, preferably 0.5 to 5% by weight when used as an internal preparation. When it is used as a coating agent, it is 0.01 to 10% by weight, preferably 0.05 to 5% by weight. When it is used as a mouthwash, 0.01 to 1% by weight, preferably 0.05 to 0%. .5% by weight.

  The anti-inflammatory agent, antioxidant and antibacterial agent of the present invention are an excipient, a disintegrating agent, a binder, a coating agent, a solvent, a fragrance, a refreshing agent, a sweetening agent, a coloring agent, depending on the dosage form and purpose of use. Surfactants, dispersants, emulsifiers, lubricants, thickeners, pH adjusters, buffers, stabilizers, preservatives, excipients and the like can be contained.

  The dosage of the anti-inflammatory agent, antioxidant and antibacterial agent of the present invention can be increased or decreased according to the symptoms of the user, purpose of use, sex, age, weight, doctor's judgment, etc. 0.5-100 mg / kg / day, preferably 5-50 mg / kg / day.

  Next, examples of the anti-inflammatory agent, antioxidant and antibacterial agent of the present invention will be described in detail, but the present invention is not limited to these examples.

Example 1
[Rheumatoid arthritis inhibitory action of the anti-inflammatory agent of the present invention]
In this example, there are three types of mangericon whole plant (raw grass) 625 mg, mangericon extract (hot water at 85 ° C., dried after 2 hours) 62.5 mg, mangericon microorganism fermented product (lactic acid bacteria fermentation) 625 mg. Samples were prepared by homogenizing each in 50 ml of physiological saline.

  Mangelicon whole plant and mannericon microbial fermented product are 50 mg / kg / day, 5 mg / kg / day, 0.5 mg / kg / day, and Mangelicon extract is 5 mg / kg / day, 0.5 mg / kg / day. Day, 0.05 mg / kg / day was orally administered to the test animals 0.1 ml 5 times a week. Solid feed (commercial product) and drinking water were freely consumed.

  MRL / 1pr mice (female, 4 weeks old) as test animals were divided into 10 groups so that the body weights were averaged at 5 weeks of age after preliminary breeding for 1 week.

  3 groups (n = 5 for each) of control group (n = 5), whole angelicon (50 mg / kg / day, 5 mg / kg / day, 0.5 mg / kg / day), mannericon extract (5 mg / kg / day) Three groups (each n = 5) of kg / day, 0.5 mg / kg / day, 0.05 mg / kg / day), mannericon microbial fermentation product (50 mg / kg / day, 5 mg / kg / day, 0. A total of 10 groups of 3 groups (5 mg / kg / day) (each n = 5), and the administration period was oral administration (5 days / week) from 5 weeks to 10 weeks of age. The breeding room was reared in an environment of 22 ± 5 ° C., humidity 50% ± 10%, and 12 hours light / dark cycle.

  The index of anti-inflammatory measurement was TNF-α. TNF-α also acts as an inflammatory cytokine involved in inflammation, pain and swelling of rheumatoid arthritis and joint destruction such as bone and cartilage. In the synovial fluid of patients with rheumatoid arthritis, TNF-α, IL-1, IL-8, etc. were excessively produced and inflammation was enhanced. TNF-α in serum collected over time from the tail vein was measured using a sandwich ELISA method. After reacting with the test mouse serum diluted 5-fold, after reacting with the antibody and coloring with the substrate solution, the absorbance at the main wavelength of 450 nm and the sub-wavelength of 620 nm was measured with a microplate reader. The measurement results are shown in FIG. In FIG. 1, * indicates that a significant difference is recognized with respect to the control (P <0.05).

  From the measurement results shown in FIG. 1, when the TNF-α value is compared with the control group as 100%, the inhibition rate in the whole angelica group is 17% in the 0.5 mg group, 31% in the 5 mg group, and in the 50 mg group. It was 36%.

  Mangericon extract also showed the same tendency, and the inhibition rate was 25% in the 0.05 mg group, 37% in the 0.5 mg group, and 44% in the 5 mg group.

  Mangericon microbial fermented product also had a suppression rate of 20% in the 0.5 mg group, 36% in the 5 mg group, and 38% in the 50 mg group.

(Example 2)
[About the inhibitory action of chemokine gene expression of the anti-inflammatory agent of the present invention]
In this example, a mangericon extract (hot water at 85 ° C., dried product extracted for 2 hours) and an extract of a mangelikon microorganism fermented product (lactic acid bacteria fermentation) (hot water at 85 ° C., dried product extracted for 2 hours) A sample was prepared in the same manner as in Example 1.

  Human fibroblasts were cultured in a 6 cm diameter culture dish until they became confluent in DMEM medium (Dulbecco's modified Eagle medium) containing 10% fetal bovine serum, and used as a test medium.

  To the test medium, the Mangelicon extract, Mangelicon microbial fermentation extract, and hydrocortisone (positive control) were added.

The final concentrations of Mangelicon extract, Mangelicon microbial fermentation extract, and hydrocortisone were 0.01% (dry weight) and 10 ⁻⁷ M, respectively. Furthermore, tumor necrosis factor TNF-α (1 ng / ml), which is known to promote chemokine gene expression, was added and cultured at 37 ° C. for 6 hours.

  Next, RNA was isolated from cells according to a conventional method, cDNA was synthesized, and then the expression level of IL-8 gene was measured by a quantitative PCR method (TaqMan PCR method). The GAPDH (glyceraldehyde 3-phosphate dehydrogenase) gene used as an internal standard gene and the obtained data corrected are shown in FIG. 2 as measurement results.

  From the measurement results shown in FIG. 2, the expression level of DMSO alone was 0.45, whereas that of Mangelicon extract was 0.27 and that of Mangelicon microbial fermented product was 0.23. It was confirmed that the gene expression of IL-8 in the cells was suppressed.

  Hydrocortisone has a strong inhibitory effect, but because it is a steroid hormone, it may cause side effects such as inflammation, tissue damage, and ulceration. On the other hand, the mangericon extract and the microbial fermented extract do not have such side effects and can be used for a long time.

  It is considered that inflammatory cells such as neutrophils and eosinophils migrate and activate in inflammatory sites such as contact dermatitis and atopic dermatitis, and are involved in the generation and amplification of inflammation. It is known that chemokines such as interleukin-8 (IL-8, interleukin-8) are produced from cells such as skin keratinocytes and fibroblasts. In psoriasis, which is an inflammatory abnormal keratosis disease, it is also known that IL-8 is present in a large amount in the affected skin area.

  Therefore, suppressing the expression of chemokines such as IL-8 is considered effective in improving or preventing inflammatory or allergic disease symptoms.

(Example 3)
[Regarding DPPH radical scavenging ability of antioxidant of the present invention]
In the present example, a MANGERICON extract (hot water at 85 ° C., dried product extracted for 2 hours), a MANGERICON extract (hot water at 40 ° C., dried product extracted overnight), an ethanol extract or 70% hydrous ethanol A sample was prepared in the same manner as in Example 1 using an extract, an extract of a fermented mangericon microorganism (hot water at 85 ° C., dried product extracted for 2 hours), an ethanol extract or a 70% aqueous ethanol extract. did.

  The DPPH radical scavenging ability was obtained by taking 1 ml of an ethanol solution in which 100 μM DPPH (1,1-Diphenyl-2-picrylhydrazyl) was dissolved in ethanol, adding 20 μl of each sample individually, and stirring the mixture at room temperature in the dark. Each sample obtained after standing for 30 minutes was measured for absorbance at a spectrophotometer of 517 nm, and a decrease in DPPH radicals was measured. The measurement results are shown in FIG.

  From the measurement results shown in FIG. 3, the DPPH radical scavenging ability of the extract of mangericon extract (hot water, hot water) and the extract of manngericon microorganism fermentation product (hot water) was low, but the antioxidant effect was confirmed.

  In addition, the DPPH radical scavenging ability of the Mangelicon ethanol extract, 70% aqueous ethanol extract, microbial fermentation ethanol extract, and 70% aqueous ethanol extract was high, and a high antioxidant effect was confirmed in a concentration-dependent manner.

Example 4
[Regarding growth inhibition and antibacterial action of the antibacterial agent of the present invention against periodontal disease bacteria]
In this example, using a MANGERICON extract (hot water at 85 ° C., dried product extracted for 2 hours) and an extract of a MANGERICON microorganism fermentation product (hot water at 85 ° C., dried product extracted for 2 hours), Samples prepared in the same manner as in Example 1 were confirmed to have antibacterial activity against periodontopathic fungi, Porphyromonas gingivalis and Actinomyces naeslundii.

  In the pure water, the mangericon extract and the mangericon microbial fermentation extract were added and dissolved so as to be 1% each. A plate medium was prepared with 10 ml GAM agar (1.5%), and 200 μl of each test bacterial solution (the above two types) / 15 ml GAM agar (1%) was layered thereon.

  Make a 2mm square hole in the cured medium at 5-7 places with a sterilized instrument. A 1% solution of mangericon extract and a 1% solution of mangericon microbial fermentation extract were added to the holes.

  Aerobic or anaerobic culture was performed at 37 ° C. for 24 hours, and the diameter of the inhibition zone after development was measured. The measurement results are shown in FIG.

From the measurement results shown in FIG. 4, it was possible to confirm the growth inhibition and antibacterial action of the extract of mangelikon extract and mangelikon microorganism fermented product, although they were two types of periodontal disease bacteria. Since antibacterial action against these periodontal disease bacteria was confirmed, it is presumed that the antibacterial agent of the present invention is also effective in preventing bad breath.

Claims (4)

  An anti-inflammatory agent characterized by having rheumatoid arthritis-inhibiting action, comprising as an active ingredient any of whole mangericon, mangericon extract, and fermented mangericon microorganism.   An anti-inflammatory agent characterized by having a chemokine gene expression-inhibiting action, comprising any one of whole mangericon, a mangericon extract, and a fermented mangericon microorganism as an active ingredient.   An antioxidant characterized by having DPPH radical scavenging ability, comprising any one of whole mangericon, mangericon extract and mangericon microorganism fermented product as an active ingredient. An antibacterial agent characterized by having an antibacterial action against periodontal disease bacteria, comprising any one of whole mangericon, mangericon extract and fermented mangericon microorganism as an active ingredient.

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