Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method of evaluating or selecting agents for preventing or improving pigmented spots

A technology of action and preparation, applied in the direction of material inspection products, measuring devices, instruments, etc.

Inactive Publication Date: 2011-02-09
KAO CORP
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] However, there are no observations that can explain the relationship between plaques and p53 activity, p53 genes and transcription targets of p53 genes (hereinafter, referred to as "p53 target genes"), or their expression products

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method of evaluating or selecting agents for preventing or improving pigmented spots
  • Method of evaluating or selecting agents for preventing or improving pigmented spots
  • Method of evaluating or selecting agents for preventing or improving pigmented spots

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] In this example, the expression of p53 in human skin tissue was analyzed by immunohistochemistry.

[0098] Skin tissue was collected from human skin with age spots, and paraffin sections of the tissue were prepared. After removal of paraffin, in REAL TM Each tissue section was heat-treated in Target Retrieval Solution (product of DAKO) at 95° C. for 45 minutes, thereby activating the antigen. After cooling at room temperature for 30 minutes, the tissue sections were rinsed with phosphate buffered saline (PBS), and then rinsed with 0.3% H 2 o 2 The solution was treated for 30 minutes. After washing with PBS, the tissue section was blocked with 10% normal goat serum (Nichirei Bioscience product) at room temperature for 1 hour, and then the mouse anti-human p53 antibody (DO-7, DAKO product) (primary antibody), and then, the sections were left overnight at 4°C. After washing with PBS, a peroxidase-labeled anti-mouse IgG polyclonal antibody (Fab') (product of Nichirei B...

Embodiment 2

[0101] In this example, the expression of p53 target genes in human epidermal tissue was analyzed.

[0102] First, the epidermis at the spot location, near the spot location (outer forearm), or sun-protected area (inner upper arm) was obtained from the skin of a healthy male adult according to the blister method. Specifically, each test site was sterilized, and the skin surface was contacted with a 1.0-2.5 mm diameter syringe (product of Terumo Corporation), followed by pumping for about 1-2 hours, and the epidermis was separated from the dermis. The isolated epidermis was isolated with sterile scissors to prepare a test body-derived sample.

[0103] Rinse the prepared epidermal tissue samples with PBS, and then transfer into RNAlater containing TM (QIAGEN product) in a test tube (1 mL), and then left to stand overnight at 4°C. Afterwards, the tissue sections were rinsed with PBS, and total RNA was prepared using the RNeasy Micro Kit (QIAGEN product) according to conventiona...

Embodiment 3

[0110] Human epidermal cells derived from the foreskin of newborns were purchased from Kurabo Industries Ltd. at 37°C, 5% (v / v) CO 2 Under atmosphere, it was precultured in a serum-free basal growth medium (EpiLife, product of Kurabo Industries Ltd.). The pre-cultured epidermal cells were inoculated into 6-well culture plates (Falcon products) (1.0 × 10 5cells / well). After 24 hours, cells were cultured in growth medium not containing BPE (bovine pituitary gland extract) and hEGF (human epidermal growth factor). After 24 hours, 5-fluorouracil (5-FU, product of Calbiochem), a generally known p53 activator, was added at a concentration of 1-10 μg / mL. After 48 hours of treatment, use Reagents (products of Invitrogen Corporation) were used to extract total RNA. Specifically, wash each well of the culture plate with PBS, add Reagent (1 mL). The mixture was poured into a 1.5 mL test tube, chloroform (200 μt L) was added thereto, and the mixture was thoroughly stirred, followe...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The object of the present invention is to provide a method for evaluating or selecting an agent having preventive or alleviative effects on spots, which method realizes reliable and effective screening of candidate substances of the agent; a method for predicting spot formation; and a method for analyzing the condition of skin spots. The method of the present invention for evaluating or selecting an agent having preventive or alleviative effects on spots, includes evaluating a substance which suppresses a p53 activity, or an expression level of a p53 gene, a p53 target gene or an expression product thereof in an epidermal cell.

Description

technical field [0001] The present invention relates to a method of evaluating or selecting an agent having a preventive or alleviating effect on plaques. Background technique [0002] Spots are a type of hyperpigmentation that occurs with aging and tend to form on areas of the skin that are exposed to the sun. Spotting is generally thought to be caused by prolonged and repeated exposure to sunlight. It has been reported that secretion of some cytokines that activate melanin synthesis in epidermal melanocytes is increased in epidermal keratinocytes at focal skin sites (Non-Patent Documents 1 to 3). [0003] So far, various research results on the mechanism of temporary pigmentation caused by ultraviolet rays have been reported (Non-Patent Document 4). On the other hand, the mechanism of chronic massive pigmentation such as age spots, which is believed to be caused by ultraviolet irradiation, has not yet been elucidated. [0004] In recent years, researchers have devoted t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50
CPCG01N33/5011G01N2333/4748
Inventor 村瀬大树八谷辉天野恭子
Owner KAO CORP
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com