JP5024506B2 - Periodontal tissue destruction inhibitor / ameliorant and oral composition - Google Patents

Description

  The present invention relates to an inhibitor / ameliorant for periodontal tissue destruction caused by endotoxin of periodontal disease bacteria and an oral composition containing the same, and an active ingredient that inhibits / improves periodontal tissue destruction caused by endotoxin of periodontal disease bacteria It relates to a screening method.

  Periodontal disease affects many people, and the prevalence of adults is particularly high. As aging progresses in the future, prevention and treatment of periodontal disease is an important issue. The main cause of this periodontal disease is bacteria in the plaque that accumulates in the periodontal pocket, and among them, anaerobic Gram-negative bacteria, especially P. gingivalis, is the cause of periodontal disease. It is attracting attention as one. When these bacteria settle and grow in the periodontal pocket, they secrete various substances and adversely affect the periodontal tissue. Among them, endotoxins (Lipopolysaccharide: hereinafter sometimes abbreviated as LPS) of these bacteria have been confirmed to cause inflammation or periodontal bone resorption directly or indirectly to the periodontal tissue. It has attracted attention in recent years.

  On the other hand, the living body tries to maintain the homeostasis of periodontal tissue by a defense mechanism by immunocompetent cells such as leukocytes and periodontal tissue constituent cells such as gingival fibroblasts. However, as periodontal disease progresses, this balance of defense mechanisms breaks down. That is, lysosomal enzymes such as cytokines, active oxygen, matrix metalloproteinases (hereinafter sometimes abbreviated as MMP) produced from the above cells become excessive, and these should originally work for a protective action. Enzymes are thought to attack and destroy their periodontal tissue.

  Conventionally, as an active ingredient for preventing and improving periodontal tissue destruction in periodontal disease, a combination of rutin or a glycoside thereof and citrate (see Patent Document 1: JP-A-7-187974), Glycyrrhetinic acid and taurine (Patent Document 2: see JP-A-8-40858), beef knee extract and leech white skin extract (Patent Document 3: see JP-A-10-167945), grape seed extract (Patent Document 4) : JP 2000-191487 A, Patent Document 5: JP 2002-29953 A), labdenic acids (Patent Document 6: JP 2005-8574 A) and the like have been proposed. However, an agent for suppressing and improving periodontal tissue destruction due to endotoxin of periodontal disease bacteria having higher efficacy is desired.

  On the other hand, the detailed mechanism of periodontal tissue destruction during periodontal disease has not been clarified. At present, MMP produced by gingival fibroblasts and immunocompetent cells for defense against infection is thought to be mainly caused by degradation of collagen, which is a major component of periodontal tissue. Inhibiting MMP production and promoting collagen synthesis are said to be effective in treating periodontal disease or preventing progression (see Patent Documents 3, 4, and 5). However, although the component disclosed by the said literatures 1-6 has the effect which improves the periodontal tissue destruction in the case of periodontal disease to some extent, it was not necessarily satisfactory.

JP-A-7-187974 JP-A-8-40858 Japanese Patent Laid-Open No. 10-167945 JP 2000-191487 A JP 2002-29953 A JP 2005-8574 A

  The present invention has been made in view of the above circumstances, clarifying the detailed mechanism of periodontal tissue destruction by endotoxin of periodontal disease, and suppressing and improving agent for periodontal tissue destruction by endotoxin of periodontal disease It is an object of the present invention to provide a composition for oral cavity, and a screening method for an active ingredient that suppresses / improves periodontal tissue destruction caused by endotoxin of periodontal disease bacteria.

  As a result of diligent studies to achieve the above object, the present inventor has eagerly pursued research to elucidate the mechanism of periodontal tissue destruction in periodontal diseases caused by periodontal endotoxin. As a result, we found that there are factors in LPS-stimulated gingival fibroblasts that increase or decrease gene expression compared to LPS-unstimulated gingival fibroblasts. To be able to screen for effective ingredients that suppress and improve periodontal tissue destruction by periodontal endotoxin, and to use this screening method to obtain an agent to suppress and improve periodontal tissue destruction by periodontal endotoxin It has been found and the present invention has been made.

Accordingly, the following invention is provided.
[1]. An agent for suppressing or improving periodontal tissue destruction by periodontal disease endotoxin comprising the following (I) or (II):
(I) Combination of lactoferrin and one or more selected from allspice extract and pine bark extract (II) Combination of lactoferrin, allspice extract and lecithin [2]. [1] A composition for oral cavity containing an agent for suppressing or improving periodontal tissue destruction by endotoxin of periodontal disease bacteria.
[3]. An oral composition containing the following (I) or (II).
(I) Combination of lactoferrin and one or more selected from allspice extract and pine bark extract (II) Combination of lactoferrin, allspice extract and lecithin [4]. The composition for oral cavity according to [2] or [3], which is toothpaste, liquid toothpaste, foam toothpaste, mouthwash, coating agent, oral paste, chewing gum, troche, gummi or candy.
[5]. The composition for oral cavity according to [2] or [3], which is a toothpaste, a liquid toothpaste, a foam toothpaste, a mouthwash, a coating agent or an oral pasta.

  ADVANTAGE OF THE INVENTION According to this invention, the suppression / improving agent excellent with respect to periodontal tissue destruction by the endotoxin of a periodontal disease microbe can be provided. In addition, it is possible to provide a simple and highly accurate screening method for selecting an active ingredient that suppresses and improves periodontal tissue destruction by periodontal disease endotoxin.

  First, the screening method of the active ingredient which suppresses and improves the periodontal tissue destruction by the endotoxin of periodontal disease bacteria is demonstrated. In this screening method, it is essential to elucidate the mechanism of periodontal tissue destruction by periodontal disease endotoxin. This mechanism was elucidated by using a periodontal tissue destruction model evaluation system as a main means of comprehensive gene expression analysis by DNA microarray analysis. That is, gingival fibroblasts are used as a model of periodontal tissue, and periodontal pathogen P. aeruginosa is one of the main causative factors of periodontal disease as a drug causing periodontal disease. A model evaluation system using gingivalis LPS was prepared and analyzed. Specifically, genes expressed in LPS-stimulated gingival fibroblasts and normal periodontal tissue LPS-unstimulated gingival fibroblasts were comprehensively analyzed using DNA microarray analysis. (For DNA microarray analysis, see "DNA microarray practice manual"). As a result, there was a group of factors that increased or decreased gene expression in gingival fibroblasts stimulated with LPS compared to gingival fibroblasts without LPS stimulation (see Test Example 1).

The following genes had increased gene expression.
MMP1: matrix metalloproteinase 1 (matrix metalloprotease 1), GeneID: 4312)
MMP3: matrix metalloproteinase 3 (matrix metalloproteinase 3), (GeneID: 4314)
MMP10: matrix metalloproteinase 10 (matrix metalloprotease 10), (GeneID: 4319)
MMP12: matrix metalloproteinase 12 (matrix metalloprotease 12), (GeneID: 4321)
FOSB: FBJ murine osteosarcoma virtual oncogene homolog B (FBJ mouse osteosarcoma viral ongogene homolog B), (GeneID: 2354)
FOS: v-fos FBJ murine osteosarcoma virtual oncogene homolog (v-fos FBJ mouse osteosarcoma viral ongogene homolog), (GeneID: 2253)
JUNB: jun B proto-oncogene (June B proto-ongogene), (GeneID: 3726)

On the other hand, the gene expression was decreased COL1A1: collagen, type I alpha 1 (type I collagen α1), (GeneID: 1277),
COL1A2: collagen, type I, alpha 2 (type I collagen α2), (GeneID: 1278),
COL3A1: collagen, type III, alpha 1 (type III collagen α1), (GeneID: 1281),
COL4A1: collagen, typeIV alpha 1 (type IV collagen α1), (GeneID: 1282),
COL4A3: collagen, type IV, alpha 3 (type IV collagen α3), (GeneID: 1285),
COL5A1: collagen, type V alpha 1 (type V collagen α1) (GeneID: 1289),
COL5A2: collagen, type V alpha 2 (type V collagen α2) (GeneID: 1290),
COL11A1: collagen, type XI alpha 1 (type XI collagen α1) (GeneID: 1301),
COL15A1: collagen, type XV alpha 1 (type XV collagen α1) (GeneID: 1306),
COL18A1: collagen, type XVIII alpha 1 (XVIII type collagen α1) (GeneID: 80781),
P4HA1: procollagen-proline, 2-oxoglutarate 4-dioxygenase proline 4-hydroxylase, alpha polypeptide I (procollagen-proline, 2-oxoglutarate 4-dioxygenase proline 4-hydroxylase, α-polypeptide ID: αGe I 5033),
P4HA2: procollagen-proline, 2-oxoglutarate 4-dioxygenase proline 4-hydroxylase, alpha polypeptide II (procollagen-proline, 2-oxoglutarate 4-dioxygenase proline 4-hydroxylase, α polypeptide II 8974),
P4HB: procollagen-proline, 2-oxoglutarate 4-dioxygenase proline 4-hydroxylase, beta polypeptide (procollagen-proline, 2-oxoglutarate 4-dioxygenase proline 4-hydroxylase, β polypeptide: 34) ,
PCOLCE: procollagen C-endpeptidase enhancer (procollagen C-endopeptide enhancer), (GeneID: 5118),
PLOD2: procollagen-lysine, 2-oxoglutarate 5-dioxygenase lysine hydroxylase2 (procollagen-lysine, 2-oxoglutarate 5-dioxygenase lysine hydroxylase 2), (GeneID: 5352),
SERPINH1: serine or cysteine proteinase inhibitor, clade H heat shock protein 47, member1, collagen binding protein1 (serine or cysteine proteinase inhibitor clade H1, protein ID1, member ID)
BMP1: bone morphogenic protein 1 (bone morphogenetic protein 1), (GeneID: 649),
ADAMTS2: a disintegrin-like and metalloprotease reprolysin type with thrombospondin type 1 motif, (GeneID: 9509).
GeneID represents a gene ID in “NCBI Entrez Gene”.

Most of the genes whose expression was increased were protein genes related to collagen degradation, and most of the genes whose expression was decreased were protein genes related to collagen synthesis. Therefore, it became clear that periodontal tissue destruction is enhanced by the following mechanism in periodontal disease.
1. Endotoxin increases the expression level of protein genes related to collagen degradation and promotes degradation of periodontal tissue.
2. Endotoxin decreases the expression level of protein genes related to collagen synthesis and weakens periodontal tissue.

  In periodontal diseases such as periodontal disease, the expression level of a protein gene related to collagen degradation increases and the expression level of a protein gene related to collagen synthesis decreases. Therefore, by controlling the expression of endotoxin inactivating agents or these genes and returning them to normal levels, it is possible to effectively suppress and improve periodontal tissue destruction caused by endotoxins of periodontal disease bacteria. .

Therefore, the following substances are effective as an agent for suppressing or improving periodontal tissue destruction by periodontal disease endotoxin. It can be set as the composition for oral cavity containing these. The oral composition will be described later.
1) Endotoxin inactivating agent.
2) A substance that returns the expression level of a gene of a protein related to collagen degradation increased by stimulation of endotoxin or a protein gene related to collagen synthesis decreased by stimulation of endotoxin to a normal level.
3) Increased by endotoxin stimulation (A) A substance that decreases the expression of a gene selected from MMP1, MMP3, MMP10, MMP12, FOSB, FOS, and JUNB.
4) Decreased by endotoxin stimulation (B) COL1A1, COL1A2, COL3A1, COL4A1, COL4A3, COL5A1, COL5A2, COL11A1, COL15A1, COL18A1, P4HA1, P4HA2, P4HB, P4HB, PICO2 A substance that increases gene expression.

  The screening method for an active ingredient that suppresses / improves periodontal tissue destruction by periodontal disease endotoxin of the present invention includes (A) a gene for a protein related to collagen degradation, which increases in periodontal diseases such as periodontal disease, Or (B) reducing the expression level of a gene of a protein related to collagen synthesis to a normal level, contacting periodontal endotoxin and a test substance with gingival fibroblasts, (A) Gene of a protein related to collagen degradation selected from the group consisting of (A) MMP1, MMP3, MMP10, MMP12, FOSB, FOS and JUNB of stimulated gingival fibroblasts, or (B) COL1A1, COL1A2, COL3A1, COL4A1 , COL4A3, COL5A1, COL5A2, COL11A1, COL15A1, COL18A1, P4 It is characterized by measuring the expression level of a protein related to collagen synthesis selected from the group consisting of A1, P4HA2, P4HB, PCOLCE, PLOD2, SERPINH1, BMP1 and ADAMTS2 and the expression level when the test substance is not contacted To do. Specifically, this expression level is compared with the expression level when the test substance is not brought into contact. (A) Increased by endotoxin stimulation (A) Decreased expression level of gene or endotoxin (B) A screening method wherein a substance whose gene expression level is increased is selected as an active ingredient for suppressing or improving periodontal tissue destruction by endotoxin of the periodontal disease bacteria.

In the screening method, periodontal disease endotoxin and a test substance are brought into contact with gingival fibroblasts. Examples of periodontal disease bacteria include P. gingivalis (hereinafter sometimes abbreviated as P. g.), A. actinomycetemcomitans (hereinafter, A. a. LPS (Lipopolysaccharide), which is an endotoxin extracted from these, is used. Gingival fibroblasts are not particularly limited, but human normal gingival fibroblasts are preferred. Periodontal disease LPS and a test substance are added to gingival fibroblasts, and cultured under conditions of 5% CO ₂ and 37 ° C. for 0.5 to 72 hours. The addition concentration of LPS of periodontal disease bacteria is preferably 1 to 100 μg / mL per 10 ⁴ to 10 ⁷ . As the medium and the like, those usually used for gingival fibroblasts are used. As described above, when LPS is added, the expression level of the (A) gene group increases and the expression level of the (B) gene group decreases compared to when LPS is not added.

  For the measurement of the gene expression level, a DNA microarray, a gene expression level quantification method using RT-PCR method, a protein expression level quantification method using antibody array, Western blotting or the like can be used. In the present invention, RNA is extracted from the cultured cells, and (A) MMP1, MMP3, MMP10, MMP12, FOSB, FOS and JUNB, and (B) COL1A1, COL1A2, COL3A1, COL4A1, COL4A3, COL5A1, COL5A2, It is preferable to measure the expression level of a gene selected from the group consisting of COL11A1, COL15A1, COL18A1, P4HA1, P4HA2, P4HB, PCOLCE, PLOD2, SERPINH1, BMP1 and ADAMTS2 with a DNA microarray.

  The expression level obtained above is compared with the expression level when the test substance is not contacted. Specifically, by this comparison, a substance that results in a decrease in the expression level of the gene group (A) increased by adding endotoxin (LPS), or decreased by adding endotoxin (LPS). (B) A substance capable of increasing the expression level of the gene group is selected as an active ingredient that suppresses / improves periodontal tissue destruction by endotoxin of periodontal disease bacteria.

  By this method, a substance having an effect of suppressing and improving periodontal tissue destruction by periodontal disease endotoxin can be easily screened with high accuracy. Substances to be screened by this screening method are not particularly limited, and examples include natural products such as plant extracts, seaweed extracts, and fungi-derived materials, chemically synthesized compounds, and genetically modified products.

  By the above screening method, the combination of lactoferrin and one or more selected from the group consisting of evening primrose extract, allspice extract, pine bark extract and lecithin was increased by adding endotoxin (LPS) ( A) Decreased expression level of the gene group, decreased by adding endotoxin (LPS), (B) Increased expression level of the gene group, and suppression and improvement of periodontal tissue destruction by periodontal disease endotoxin The effect was confirmed. The combination of lactoferrin and one or more selected from the group consisting of evening primrose extract, allspice extract, pine bark extract and lecithin is increased by an endotoxin inactivating agent, endotoxin (LPS) (A It is also useful as a gene expression suppressor or expression level reducing agent, or a gene expression promoter or expression level increasing agent that is decreased by endotoxin (LPS). It is also useful as an expression control agent for the gene (A) that is increased by endotoxin (LPS) and the gene (B) that is decreased by endotoxin (LPS).

  The lactoferrin used in the present invention is commercially available lactoferrin, mammals (eg, humans, cows, sheep, goats, horses, etc.) colostrum, transitional milk, regular milk, terminal milk, etc. or skim milk that is a processed product of these milks, Lactoferrin isolated from whey and the like by conventional methods (for example, ion exchange chromatography) and lactoferrin produced from plants (tomato, rice, tobacco). Lactoferrin may be a commercially available product or can be prepared and used by a known method. Lactoferrin is preferably derived from cattle.

  Evening primrose extract is obtained by extracting seeds of evening primrose with a solvent and contains a large amount of polyphenols such as proanthocyanidins. The allspice extract is obtained by extracting allspice fruit with a solvent. Pine bark extract is obtained by using European red pine as a raw material and extracting the bark with a solvent, and contains a large amount of proanthocyanidins, which are a kind of polyphenols such as eugeniin. As these plant extracts, commercially available products or those obtained by known methods can be used. In this case, examples of the solvent used for extraction include water; alcohols such as methanol, ethanol, propanol and butanol; and polyhydric alcohols such as propylene glycol and butylene glycol. These may be used alone or in combination of two or more. It can be used as a mixed solvent.

  Various conditions in the extraction step are not particularly limited, but usually the ratio of the extraction raw material to the extraction solvent is preferably in the range of extraction raw material: extraction solvent = 1: 5 to 1: 100 in terms of mass ratio. Examples of the extraction treatment include cold-dipping, hot-dipping, heating reflux, percolation, and the like, and it is preferable that the extraction treatment is performed by immersion or stirring in an extraction solvent for 3 hours to 2 weeks. The extraction pH is not particularly limited as long as it is not extremely acidic or alkaline. In addition to solvent extraction, an extract obtained by steam distillation or supercritical extraction performed with carbon dioxide gas in a critical state can be used as well. In supercritical extraction, hexane, ethanol, or the like can be used as an extraction aid.

  When the extraction solvent is a non-toxic solvent such as water, ethanol, water / ethanol (hydrous ethanol) or the like, the extract may be used as it is or can be used as a diluent. In addition, the extract may be a concentrated extract, which may be formed into a dry powder by freeze drying or the like, or prepared in a paste form. When other solvents are used, it is desirable to dilute the dried portion with a non-toxic solvent after distilling off the solvent.

  Lecithin is widely distributed in the living bodies of plants, animals, and microorganisms, and is extracted and separated from these living bodies to produce products. Mainly, soybean lecithin and egg yolk lecithin are commercially available. The lecithin used in the present invention is particularly preferably derived from soybean.

  The composition for oral cavity of the present invention is a periodontal tissue disruption caused by endotoxin of periodontal disease bacteria consisting of lactoferrin and one or more selected from the group consisting of evening primrose extract, allspice extract, pine bark extract and lecithin. Contains an inhibitor / improving agent.

  The content of lactoferrin is preferably 0.01 to 10% (solid mass%) in the oral composition, and more preferably 0.1 to 4% (solid mass%). Within this range, suppression and improvement of periodontal tissue destruction by endotoxins of periodontal disease bacteria are particularly effective. The content of one or more kinds selected from the group consisting of evening primrose extract, allspice extract, pine bark extract and lecithin is preferably 0.01 to 10% (solid content mass%) in the oral composition. Preferably it is 0.1 to 4%. If the content is less than 0.01%, a satisfactory effect may not be obtained. Even if the content exceeds 10%, the effect is not improved, a problem occurs in flavor, and the composition is stable. May cause problems with sex.

  As an oral composition, chewing gum is effective. Other oral compositions such as toothpaste, liquid toothpaste, foam toothpaste, mouthwash, coating agent, troche, gummi, candy, etc., and oral medicine such as tablets, capsules, powders, drinks, etc. It is. Further, depending on the application mode, dosage form, etc. of the composition, for example, surfactant, cleaning agent, moisturizing agent, thickener, abrasive, binder, thickener, oil, alcohols, polymer Substance, preservative, inclusion compound, antioxidant / antioxidant, chelating agent, inorganic powder, gum base, acidulant, softener, colorant, brightener, emulsifier, sweetener, pH adjuster, fragrance, pigment Herbal medicines, saccharides, salts, amino acids, medicinal ingredients other than those mentioned above, antibacterial agents, water and the like can be blended, and can be prepared by ordinary methods.

  As described above, the agent for suppressing and improving periodontal tissue destruction according to the present invention is extremely effective for suppressing and improving periodontal tissue destruction in periodontal diseases. In addition, the screening method of the present invention is effective in selecting an active ingredient that suppresses / improves periodontal tissue destruction caused by endotoxin of periodontal disease bacteria.

  EXAMPLES Hereinafter, although a test example, an Example, and a comparative example are shown and this invention is demonstrated concretely, this invention is not restrict | limited to the following Example. The followings were used for the lactoferrin, evening primrose extract, allspice extract, pine bark extract, lecithin and grape seed extract used in the test examples and examples.

Lactoferrin: Trade name Lactoferrin (Wako Pure Chemical Industries, Ltd.)
Evening Primrose Extract: Product Name Evening Primrose Extract P (Oryza Oil Co., Ltd.)
Pine bark extract: Brand name Flavangenol (manufactured by Toyo Shinyaku Co., Ltd.)
Lecithin: Lecithin FA (made by Honen Corporation)
Allspice extract: Allspice fruit was dried and pulverized to obtain a crude powder, and 10 g of the crude powder was immersed in 100 mL of ethanol and extracted at room temperature for 5 days. The extract obtained by filtering off the all spice fruit crude powder was concentrated under reduced pressure to obtain an extract.

[Test Example 1]
Effect of endotoxin (LPS) derived from P. gingivalis on human normal gingival fibroblasts (HGF) (1) Method 10% fetal bovine serum (FBS) in a 75 cm ² tissue culture flask Human gingival fibroblasts (HGF) suspended in Dulbecco's Modified Eagle's Medium (DMEM) containing the above were seeded (5 × 10 ⁵ cells / flask) and cultured in a CO ₂ incubator at 37 ° C. When the cells are confluent, 1% FBS and 1 μg / mL P.I. It transferred to DMEM containing LPS derived from gingivalis (ATCC 33277) and cultured for 72 hours. Controls were cultured in DMEM containing only 1% FBS. After the culture, the cells were collected, RNA was extracted with RNeasy Mini Kit (QIAGEN), and the gene expression level was analyzed using GeneChip Human Genome Focus Array (Affymetrix).

(2) Results Data on genes involved in collagen metabolism were extracted from the results of GeneChip analysis and summarized. As a result, in gingival fibroblasts stimulated with LPS, there was a group of factors in which gene expression was increased or decreased compared to LPS-unstimulated gingival fibroblasts. The results are shown in Table 1.

[Test Example 2]
A normal gingival fibroblast (HGF) suspended in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS) was seeded (5 × 10 ⁵ ) in a 75 cm ² tissue culture flask. cell / flask). P. gingivalis-derived LPS (1 μg / mL), lactoferrin, and one or more selected from the group consisting of evening primrose extract, allspice extract, pine bark extract and lecithin are mixed, and the mixture is mixed at 37 ° C. for 30 minutes. Incubated. DMEM containing this mixture and 1% FBS was prepared and added to HGF when cells were confluent. The amounts used of lactoferrin, evening primrose extract, allspice extract, pine bark extract and lecithin were adjusted to 1000 ppm each when used alone, and to 1000 ppm when two or more types were used. Two hours after the addition of DMEM containing the mixture and 1% FBS, the medium was changed to DMEM containing 1% FBS, and the culture was further continued for 18 hours. The gene expression levels of (A) MMP1, MMP3, (B) COL1A1, COL1A2, P4HA1, P4HA2, P4HB, and SERPINH1 after culture were measured by RT-PCR. The gene expression level when cultured in a LPS-free medium and the gene expression level when cultured in a medium supplemented with LPS alone (stimulated with LPS) were measured in the same manner. 1) The expression improvement rate of the gene (B) decreased by stimulation of endotoxin was calculated based on the formula (2), and was shown by the following evaluation criteria. The results are shown in Table 2.

Standard ◎: Gene expression improvement rate of 70% or more ○: Gene expression improvement rate of 50% or more △: Gene expression improvement rate of 30% or more ×: Gene expression improvement rate of less than 30%

  The culture supernatant after the culture was collected, and the amount of type I collagen and the amount of MMP-1 were measured by ELISA. Results are shown below. When cultured in a medium not added with LPS, the amount of type I collagen and the amount of MMP-1 when cultured in a medium supplemented with LPS alone (stimulated with LPS) were measured in the same manner, and the collagen amount improvement rate based on the following formula (3) Based on (%) and the following formula (4), the MMP-1 production inhibition rate (%) was calculated. The results are shown in Tables 3 and 4 and FIGS.

  From the above results, lactoferrin and one or more combinations selected from evening primrose extract, allspice extract, pine bark extract and lecithin are attributed to endotoxin (LPS) by improving the expression of the above gene group. The effect of improving the collagen synthesis inhibition and the collagen degradation promotion of the cells to be observed was confirmed.

[Prescription Examples 1-16, Reference Examples 1-16]
The formulation example of the composition for oral cavity of this invention is shown. Each formulation example was prepared according to the usual method of each dosage form according to the composition.

6 is a graph showing a collagen amount improvement rate in Test Example 2. 5 is a graph showing the MMP-1 production inhibition rate in Test Example 2.

Claims (5)

An agent for suppressing or improving periodontal tissue destruction by periodontal disease endotoxin comprising the following (I) or (II):
(I) A combination of lactoferrin and one or more selected from allspice extract and pine bark extract (II) A combination of lactoferrin, allspice extract and lecithin   The composition for oral cavity containing the inhibitor of the periodontal tissue destruction by the endotoxin of the periodontal disease bacteria of Claim 1, and an improving agent. An oral composition containing the following (I) or (II).
(I) A combination of lactoferrin and one or more selected from allspice extract and pine bark extract (II) A combination of lactoferrin, allspice extract and lecithin   The composition for oral cavity according to claim 2 or 3, which is toothpaste, liquid toothpaste, foam toothpaste, mouthwash, coating agent, oral paste, chewing gum, troche, gummi or candy.   The composition for oral cavity according to claim 2 or 3, which is a toothpaste, liquid toothpaste, foam toothpaste, mouthwash, coating agent or oral paste.

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