JP6188986B1 - Clock gene expression level regulator and elastin production promoter - Google Patents

Abstract

An object of the present invention is to provide a clock gene expression level regulator that regulates the expression level of a clock gene, and to provide an elastin production promoter that promotes the production of elastin. [MEANS FOR SOLVING PROBLEMS] A clock gene expression level regulator comprising the honeysuckle extract of the present invention as an active ingredient can exhibit a high clock gene expression level regulation effect. Moreover, the elastin production promoter which uses the honeysuckle extract of this invention as an active ingredient can exhibit the high elastin production promotion effect. [Selection] Figure 1

Description

  The present invention relates to a clock gene expression level adjusting agent and an elastin production promoter comprising a honeysuckle extract as an active ingredient.

  The circadian rhythm is a rhythm with a period of about 24 hours that a living organism has, and is controlled by a biological clock possessed by the organism. The circadian clock causes biological diurnal fluctuations and controls the diurnal fluctuations of various biological activities such as body temperature, blood pressure, hormone secretion, metabolism, as well as mental and physical activities, contact, including sleep and awakening. It is thought that In recent years, disturbance of circadian rhythm has been pointed out as a cause of various psychosomatic symptoms or diseases such as sleep disorders, skin diseases, lifestyle-related diseases, and neuropsychiatric diseases such as depression. In addition, the clock gene is expressed in the skin, and the homeostasis is maintained by rhythming the clock gene in synchronism with the central clock in the brain, as in other organs.

  The circadian clock is controlled by a circadian clock system that includes a group of genes called clock genes. Among clock genes, the BMAL1 gene, in particular, is a basic helix-loop-helix protein that forms a heterodimer with the CLOCK gene, and is known to be highly expressed at night and low in expression during the day.

  It is also known that clock genes regulate skin homeostasis. (Non-patent document 1.)

  In order to maintain the body clock normally, those that regulate the expression of clock genes involved in the body clock system are effective. For example, Patent Document 1 discloses a clock gene expression promoter containing lavender oil or eucalyptus oil and an alkylene oxide derivative. Patent Document 2 discloses a circadian rhythm improving agent containing a treated product of specific lactic acid bacteria as an active ingredient.

  Elastin is present in articular cartilage and ligaments, and helps smooth operation of joint functions. In the case of osteoarthritis (OA), the blood circulation around the joints deteriorates and the supply of oxygen decreases, thereby reducing the production of collagen and proteoglycans by chondrocytes, and dead chondrocytes stimulate the synovium and inflammation Cause pain in the joints. In addition, the release of cytokines during inflammation induces chondrocyte death and intensifies pain symptoms. Thus, an elastin production promoter has been desired.

  Symptomatic treatment of OA includes administration of pain relievers and anti-inflammatory preparations, injection of high molecular hyaluronic acid (sodium hyaluronate) into the joint cavity, and the like. In recent years, approaches other than symptomatic treatment such as promotion of chondrocyte differentiation or suppression of chondrocyte hypertrophy, chondrocyte-mediated collagen and proteoglycan transcription, synthesis promotion, etc. have begun to be taken. There are not many studies.

  As elastin production promoters, extracts from plants such as Kyomizuna, Kyojou Nana, Fushimi Togarashi, Manganji Togarashi and Kujo Negi, Ogaku, Ginseng, Altea, Yokuinin and Lily are disclosed. (Patent Document 3)

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Cosmetology Research Report Vol.24, 2016, 117-123p.

Japanese Patent Publication No. 2013-46272 JP 2013-181005 A JP2012-162487

  An object of the present invention is to provide a clock gene expression level regulator that regulates the expression level of a clock gene, and to provide an elastin production promoter that promotes the production of elastin.

  The clock gene expression level regulator comprising the honeysuckle extract of the present invention as an active ingredient can exert a high clock gene expression level regulation effect.

  The elastin production promoter comprising the honeysuckle extract of the present invention as an active ingredient can exhibit a high elastin production promotion effect.

FIG. 1 is a diagram showing that the expression level of BMAL1 gene increases at night by adding honeysuckle extract. FIG. 2 is a diagram showing that the expression level of the elastin production gene increases at night by adding the honeysuckle extract.

  Hereinafter, modes for carrying out the present invention will be described.

The honeysuckle extract used in the present invention may be an extract obtained from a plant of the genus Caprifoliaaceae ( Lonicera L.). The honeysuckle plant, honeysuckle, warbler Kagura (Lonicera japonica Thunb.); Uguisunoki, Hana gourd I (Lonicera maackii (Rupr) Maxim. .), Gourd I (Lonicera gracilipes Miq var glabra Miq. ..); Lonicera Morrowii (Lonicera morrowii A. Gray), smell caulis lonicerae. (Lonicera periclymenum L.), luck Nuqui caulis lonicerae (Lonicera sempervirens L.), chromium Roh warbler Kagura;.. chromium Roh warbler, black warbler (Lonicera caerulea subsp edulis (Turcz) Hulten var emphylloc (. Maxim) alyx Nakai), Arage gourd me;... Oba gourd I (Lonicera strophiophora Franch), mountain warbler Kagura (Lonicera gracilipes Miq var gracilipes), Miyama warbler Kagura (Lonicera gracilipes Miq var glandulosa Maxim) ,... co-warbler Kagura (Lonicera ramosissima Franch. et Sav. ex Maxim.), small basket gourd I (Lonicera linderifolia Maxim. var. konoi (Makino) Okuyama), privet gourd I (Lonicera demissa Rehder), Chishimahyoutanboku (Lonicera chamissoi Bunge), thin blade gourd I (Lonicera cerasina Maxim.), Demon gourd I (Lonicera vidalii Franch. Et Sav. ), Giant gourd I (Lonicera tschonoskii Maxim.), Nikko gourd I (Lonicera mochidzukiana Makino), Ezo gourd I (Lonicera alpigena L. subsp glehni (F. Schmidt) H Hara..), Kidachinindou; Tounindou, Korean caulis lonicerae (Lonicera hypoglauca Miq. ), Hamanindou (Lonicera affinis Hook. Et Arn. ), Smell caulis lonicerae (Lonicera periclymenum), Ronikera-Etorusuka (Lonicera etrusca), Nemurobushidama (Lonicera chrysantha var. Crassipes), Ha Yazaki gourd I (Lonicera praeflorens var. Japonica), safflower gourd I (Lonicera maximowiczii var. sachalinensis), Suruga gourd I (Lonicera alpigena ssp. glehnii var. viridissima), Ronikera-Arupigena (Lonicera alpigena ssp. alp igena), Keyonomi (Lonicera caerulea ssp. edulis), Marubayonomi (Lonicera caerulea ssp. edulis var. venulosa), chromium Roh warbler Kagura (Lonicera caerulea ssp. edulis var. emphyllocalyx), Europe Yo flea (Lonicera caerulea ssp. caerulea), etc. It has been known. The plant used as a raw material used in the present invention is not particularly limited as long as it is a honeysuckle genus plant, but honeysuckle ( Lonicera japonica Thunb.) Is preferably used because of its relatively easy availability and effectiveness. For extraction, each part of the plant such as flowers, leaves, stems, branches and fruits can be used. In the present invention, flowers are preferably used from the viewpoint of effectiveness.

  When extracting the honeysuckle extract used in the present invention, the honeysuckle extract may be used as it is, but considering the extraction efficiency, it is preferable to carry out the extraction after performing processing such as shredding, drying, and grinding. The extraction can be performed by immersing in an extraction solvent or by an extraction method using a supercritical fluid or a subcritical fluid. In order to increase the extraction efficiency, homogenization may be performed in stirring or an extraction solvent. The extraction temperature is suitably about 5 ° C. to the boiling point of the extraction solvent. The extraction time varies depending on the type of extraction solvent and the extraction temperature, but it is appropriate to set it to about 1 hour to 14 days.

  Extraction solvents include water, lower alcohols such as methanol, ethanol, propanol, and isopropanol, polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin, and ethers such as ethyl ether and propyl ether. , Polar organic solvents such as esters such as butyl acetate and ethyl acetate, and ketones such as acetone and ethyl methyl ketone can be used, and one or more of these are selected and used. Further, physiological saline, phosphate buffer, phosphate buffered saline, or the like may be used. Furthermore, you may use 1 type, or 2 or more types of supercritical fluids and subcritical fluids, such as water, a carbon dioxide, ethylene, propylene, ethanol, methanol, ammonia. Among these extraction solvents, it is preferable to use one or two selected from water and ethanol from the viewpoint of the effect of the present invention.

  Honeysuckle extract with the above-mentioned solvent can be used as it is, but it is decolorized and deodorized as long as the concentrated and dried solids are dissolved again in water or a polar solvent or their skin physiology is not impaired. , And may be used after a purification treatment such as desalting or a fractionation treatment by column chromatography. For storage, it can be freeze-dried after purification and dissolved in a solvent before use. It can also be used by encapsulating in vesicles such as liposomes or microcapsules.

  The honeysuckle extract exhibits an excellent clock gene expression level adjusting effect and can be used as a clock gene expression level adjusting agent.

  The honeysuckle extract exhibits an elastin production promoting effect by increasing the expression of the elastin production gene, and can be used as an elastin production promoter.

  EXAMPLES Hereinafter, the present invention will be specifically described with reference to examples, but the scope of the present invention is not limited thereby.

[Honeysuckle extract]
Honeysuckle (Lonicera japonica) flowers were dried and then chopped and immersed in a 20-mass volume ethanol / purified water mixture (1: 3), and the filtrate after filtration was collected to obtain a honeysuckle extract.

[BMAL1 gene expression regulating action, ELASTIN gene expression promoting action]
Human fibroblasts were seeded in a 6-well plate at 5 × 10 ⁵ cells / well and cultured overnight in DMEM medium containing 1% FBS. The honeysuckle extract was replaced with a dissolved medium, and cultured in a 37 ° C., 5% CO ₂ incubator for 24 hours. Total RNA extraction from cells was prepared using Trizol reagent according to the protocol of Life Technologies. After cDNA synthesis, gene expression was confirmed by real-time PCR by the cyber green method using the following primers. GAPDH was used as an internal standard. The results are shown in Table 2.

  As shown in FIG. 1, the expression level of the BMAL1 gene, which is a type of clock gene, was changed by adding the honeysuckle extract. In particular, the expression level of the BMAL1 gene markedly increased in 8 and 12 corresponding to nighttime. In contrast, no significant change was observed in the expression level of the BMAL1 gene at 0, 2, 4, and 24 corresponding to daytime. Therefore, it was revealed that the expression level of the clock gene is adjusted by adding the honeysuckle extract.

  As shown in FIG. 2, the addition of the honeysuckle extract tended to increase the expression of the elastin gene, and a remarkable increase was observed particularly at 4 and 8 corresponding to nighttime. Therefore, it became clear that the expression level of an elastin production gene increases by adding a honeysuckle extract.

Claims (1)

  A BMAL1 gene expression level regulator comprising honeysuckle extract as an active ingredient.

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