JP2019052145A5 - - Google Patents

Description

^{(Test Example 5) Evaluation of MITOL gene expression promoting action on dermal fibroblasts 1.0 × 10 5} cells / mL of dermal fibroblasts (manufactured by Kurabo Industries) in medium FibroLife BM (manufactured by Kurabo Industries, with attached supplement added) The cells were adjusted to cell density, 0.5 mL each was seeded on a 24-well plate, and cultured in an incubator set at 37 ° C and 5% CO2. The next day, the medium was removed by suction, replaced with medium FibroLife BM (manufactured by Kurabo Industries, supplements other than FBS and FGF were added), and cultured for another 24 hours. The medium was removed by suction, replaced with a medium containing the test substance, and cultured for 24 hours. After completion of the culture, the medium was removed, lysate buffer was added to lyse the cells, and the cytolytic solution was collected. RNA was recovered from the cell lysate according to the attached protocol using RNeasy Mini Kit (Qiagen), and cDNA was synthesized by reverse transcription reaction using Prime Script RT Master mix (Takara Bio) using this as a template. From the synthesized cDNA, the expression levels of GAPDH and MITOL mRNAs were measured by a real-time PCR system (Step One Plus, Thermo Fisher Scientific) (SYBR Green method), and the expression level of MITOL was corrected by the GAPDH expression level. .. As the primer, the one of the following model number was used. GAPDH: HA067812, MITOL: HA192576 (both are Takara Bio). The measured mRNA expression level was compared with the control group (medium containing no test substance), and the MITOL gene expression promoting effect of the test substance was evaluated.
<Test results>
The results are shown in FIG. Fermented rice bran extract, Oubaku extract, Ouren extract, Peach extract, Tamogitake extract, β-estradiol, 5-aminolevulinic acid, β-nicotinamide mononucleotide, riboflavin, thiotaurine, MA-5, metformin, urolithin A, niacin It was confirmed that amide and D-panthenol increased the mRNA expression level of MITOL. These substances can be used as positive controls when screening for components that promote MITOL production.

^{(Test Example 8) Evaluation of the effect of the material on gene expression in dermal fibroblasts 1.0 × 10 5} cells / mL of dermal fibroblasts (manufactured by Kurabo Industries) in medium FibroLife BM (manufactured by Kurabo Industries, with attached supplement added) The cells were seeded at 0.5 mL each on a 24-well plate and cultured for 24 hours in an incubator set at 37 ° C and 5% CO2. The medium was removed by suction, replaced with medium FibroLife BM (manufactured by Kurabo Industries Ltd., supplements other than FBS and FGF were added), and cultured for another 24 hours. The medium was removed by suction, replaced with a medium containing the test substance, and cultured for 24 hours. After completion of the culture, the medium was removed, lysate buffer was added to lyse the cells, and the cytolytic solution was collected. RNA was recovered from the cell lysate according to the attached protocol using RNeasy Mini Kit (Qiagen), and cDNA was synthesized by reverse transcription reaction using Prime Script RT Master mix (Takara Bio) using this as a template. From the synthesized cDNA, the expression levels of GAPDH, LOXL-1 and MME mRNAs were measured by a real-time PCR system (Step One Plus, Thermo Fisher Scientific) (SYBR Green method), and the expression of LOXL-1 and MME was measured. The amount was corrected by the expression level of GAPDH. As the primer, the one of the following model number was used. GAPDH: HA067812, LOXL-1: HA230268 and MME: HA211733 (both are Takara Bio). The measured mRNA expression level was compared with the control group (medium containing no test substance), and the effect of promoting various gene expressions by the test substance was evaluated.
<Test results>
The results are shown in FIG. It was confirmed that the mRNA expression level of LOXL-1 was increased by the Pleurotus cornucopiae extract and the β-nicotinamide mononucleotide. It was also confirmed that the peach extract and riboflavin reduced the mRNA expression level of MME. From the above, the component that promotes MITOL production can be utilized as a production promoter of the factors listed in Table 1 and a production inhibitor of the factors listed in Table 2. Further, when screening the production promoter of the factors shown in Table 1 and the production inhibitor of the factors shown in Table 2, the MITOL production promoting action can be used as an index.

Claims (14)

Buttonpi extract, peach extract, Hamamelis extract, niacinamide, Oubaku extract, Ouren extract, Komenuka fermented extract, Senburi extract, Eucalyptus extract, Tamogitake extract, β-estradiol, 5-aminolevulinic acid, β-nicotin An MITOL reduction inhibitor or production promoter comprising at least one selected from the group consisting of amide mononucleotide, riboflavin, thiotaurine, MA5, metformin, urolithin A, and D-pantenol. A composition for promoting the production of the factors shown in Table 1 or for suppressing the production of the factors shown in Table 2, which contains a component that promotes MITOR production as an active ingredient. Ingredients that promote MITOR production are fermented rice bran extract, aubergine extract, auren extract, semburi extract, hamamelis extract, buttonpi extract, peach extract, eucalyptus extract, tamogitake extract, β-estradiol, 5-aminolevulin. The composition according to claim 2 , which is at least one selected from the group consisting of acid, β-nicotinamide mononucleotide, riboflavin, thiotaurine, MA5, metformin, urolithin A, niacinamide, and D-pantenol. The composition according to claim 2 , which is used for suppressing, promoting or activating the decrease of hair follicle stem cells, pigment stem cells, hair follicles or epidermal keratinocytes, or dermal fibroblasts. The composition according to claim 2 , wherein the factors shown in Table 1 are COL17A1, SOD2, LOXL-1. The composition according to claim 2 , wherein the factor shown in Table 1 is COL17A1. The composition according to claim 6 , which is used for preventing or improving white hair. The composition according to claim 2 , wherein the factors shown in Table 1 are COX4I1, TNNI1, MB. A composition for promoting slowing muscles or improving muscle endurance, which comprises a witch hazel extract or a eucalyptus extract. Factor according to Table 2 is a MME, the composition according to claim 2. A method for suppressing the decrease of hair follicle stem cells, pigment stem cells, hair follicles or epidermal keratinized cells, or dermal fibroblasts, promoting proliferation, or screening an activator, using the MITOL production promoting action as an index. A method for suppressing a decrease in COL17A1, SOD2, LOXL-1, COX4I1, TNNI1 or MB or screening a production promoter using the MITOL production promoting action as an index. A method for screening an agent for promoting MME reduction or suppressing production, using the MITOL production promoting action as an index. A method for screening a production promoter of the factors shown in Table 1 or a production inhibitor of the factors shown in Table 2 using the MITOL production promoting action as an index.