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Detection kit for skin-active ingredients and method for detecting skin-active ingredients by using the same

A technology for detecting kits and active substances, used in biochemical equipment and methods, testing pharmaceutical preparations, measuring devices, etc.

Inactive Publication Date: 2012-05-23
AMOREPACIFIC CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there are no reports of studies on substances that increase the expression of klotho

Method used

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  • Detection kit for skin-active ingredients and method for detecting skin-active ingredients by using the same
  • Detection kit for skin-active ingredients and method for detecting skin-active ingredients by using the same
  • Detection kit for skin-active ingredients and method for detecting skin-active ingredients by using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0020] According to one embodiment of the present invention, the skin cells may be human keratinocytes, fibroblasts or melanocytes, but are not limited thereto.

[0021] In addition, the present invention relates to the detection of skin active substances through the steps of determining whether klotho protein is expressed or not and the step of isolating and refining substances that can increase the expression of klotho protein after treating skin cells with candidate substances using the detection kit Methods.

[0022] According to another embodiment of the present invention, using the detection kit, it is determined that retinoic acid and retinol, which are known as antioxidant substances, can increase the expression of klotho protein, thereby determining that the detection method according to the present invention is suitable for screening unknown Anti-aging substances and skin active substances.

[0023] Hereinafter, the contents of the present invention will be describe...

experiment example 1

[0024] Experimental Example 1 Determination of klotho protein expression in skin cells

[0025] In order to determine whether the klotho protein is expressed in skin cells, primers capable of amplifying the klotho gene (NCBI record number: NM_004795) with the sequences shown in SEQ ID No: 1 and SEQ ID No: 2 were prepared, and the primer pair and a probe having the sequence shown in SEQ ID No: 3 (TaqMan TM Fluorescent probe), observed by RT-PCR in keratinocytes (NHK), fibroblasts (NHF), NHM-enhanced black (NHM-promoblack) cells (Normal Human Epidermal Melanocytes, NHEM) primary Foreskin, Negro-derived melanocytes, Promocells), NHM-DP cells (NHM-dark pigment; Negroid-derived melanocytes, Invitrogen) and NHM-MP cells (NHM-Medium Pigment (NHM-moderate pigment); whether the mRNA of klotho can be detected in the cDNA of Asian melanocytes, Invitrogen). At this time, the PCR reaction conditions were denaturation at 94°C for 30 seconds, annealing at 55°C for 30 seconds, and extension...

experiment example 2

[0027] Experimental Example 2 Measurement of klotho mRNA expression in keratinocytes treated with retinoic acid and retinol

[0028] In order to determine the effects of retinoic acid and retinol, which are known as anti-aging substances, on the expression of klotho in keratinocytes, RT-PCR was carried out under the same conditions as in Experimental Example 1, using the retinoic acid 0.1 and 1 μM and retinol 1 and 10 μM, the expression of klotho protein was measured on day 2 and day 8, and the results were reported in image 3 Indicated.

[0029] from image 3 It can be seen from the results that the expression of klotho protein significantly increased on the 8th day after treatment with retinoic acid and retinol.

[0030]

[0031]

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Abstract

The invention relates to a detection kit for skin-active ingredients and a method for detecting skin-active ingredients by using the same. Specifically, the invention relates to a detection kit for skin-active ingredients comprising klotho genes known as longevity genes. Candidate substances for inducing klotho gene expression from skin cells are separated and refined, thereby detecting anti-aging substances, whitening substances and other skin-active ingredients.

Description

technical field [0001] The invention relates to a kit for detecting skin active substances and a method for detecting skin active substances using the kit, in particular to using a detection kit containing the klotho gene known as the longevity gene to induce the klotho gene from skin cells A kit for detecting skin active substances such as skin active substances such as anti-aging and whitening substances by separating and refining expressed candidate substances, and a method for detecting skin active substances using the kit. Background technique [0002] Skin ages with age or with exposure to sunlight. With aging, the cuticle of the epidermis takes longer to exfoliate and renew, the cuticle thickens, and the dermis becomes wrinkled due to the decrease in the content or arrangement of elastic fibers and moisturizing ingredients. So far, various studies have been conducted on the prevention of skin changes caused by ultraviolet irradiation in order to block photoaging, whi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/68C12Q1/6883G01N33/6881C12Q2600/158G01N33/5044C12Q2600/148G01N33/15G01N33/5023C12Q1/6876G01N33/5302
Inventor 崔向兑孙义东李真锳罗勇柱金汉坤
Owner AMOREPACIFIC CORP
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