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Method for stimulation collagen synthesis and/or kgf expression

A collagen synthesis and composition technology, which is applied in the direction of drug combination, pharmaceutical formula, skin care preparation, etc., can solve the problem of not being identified

Inactive Publication Date: 2008-01-23
伊玛吉恩有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the relationship between AIMP1 and collagen synthesis and KGF expression has not yet been clarified.

Method used

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  • Method for stimulation collagen synthesis and/or kgf expression
  • Method for stimulation collagen synthesis and/or kgf expression
  • Method for stimulation collagen synthesis and/or kgf expression

Examples

Experimental program
Comparison scheme
Effect test

reference example 1

[0062] Construction of AIMP1 and fragments

[0063] AIMP1 protein (SEQ ID NO: 1) consisting of 312 amino acids, its N- Terminal fragment (1-147; SEQ ID NO: 2) and C-terminal fragment (148-312; SEQ ID NO: 3).

reference example 2

[0065] Construction of N-terminal deletion fragment and C-terminal deletion fragment of AIMP1

[0066] N-terminal deletion fragments and C-terminal deletion fragments of AIMP1 were prepared respectively, namely, AIMP1-(6-46; SEQ ID NO: 12), AIMP1-(1-46; SEQ ID NO: 13), AIMP1-(1-53; sequence No. 14), AIMP1-(193-312; Sequence No. 15), AIMP1-(1-192; Sequence No. 27) and AIMP1-(6-192; Sequence No. 28) fragments. The above-mentioned fragments all use the cDNA of AIMP1 as a template, and use the specific primer set (Table 1) for each fragment to carry out PCR amplification and synthesis for each fragment. The PCR reaction conditions are as follows: heat at 95°C for 2 minutes to pre-denature the template DNA; then perform 25 cycles as follows: 30 seconds at 95°C; 30 seconds at 56°C; and 1 minute at 72°C; The extension reaction was carried out at 72°C for 5 minutes. The above PCR product was digested with EcoRI and XhoI, and ligated into pGEX4T3 vector (Amersham Biosciences) diges...

Embodiment 1

[0070] Stimulation of collagen synthesis by different concentrations of AIMP1

[0071] Foreskin fibroblast culture and AIMP1 treatment

[0072] In order to measure the collagen transcription RNA induced by AIMP1, foreskin fibroblasts (5×10 4 Cells / well, company name: MTT, deposit number: MC1232) were cultured on a 6-well plate in DMEM medium containing 10% serum for 12 hours, and then cultured in a serum-free medium for about 3 hours. Then, the cultured cells were treated with different concentrations of AIMP1 protein (SEQ ID NO: 1) (0, 20, 50, 100 and 200 nM) for 6 hours (RT-PCR) or 12 hours (immunoblotting).

[0073] RT-PCR analysis

[0074] After the cells treated with different concentrations of AIMP1 in the above-mentioned Example were recovered, they were dissolved with TRIzol (invitrogen). 10 wt% chloroform was added to the cell lysate and mixed well. The mixture was centrifuged at 12,000 g for 15 minutes, and the supernatant was taken. Ethanol was added to the...

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Abstract

The present invention relates to a method for stimulating collagen synthesis and / or KGF expression, and more particularly, to a method for stimulating collagen synthesis and / or KGF expression using the AIMPl or its fragment. The AIMPl or its fragment can be effectively used for the stimulation of collagen synthesis and / or KGF expression in a subject in need thereof, the treatment of skin aging in the subject, the treatment of the flaccid and / or wrinkled skin in the subject, the promoting the smoothing and / or firming of the skin in the subject, the treatment of adverse cutaneous effects of menopause in the subject, and the treatment of adverse effects of menopause on the collagen.

Description

technical field [0001] This application claims priority from patent application number PCT / KR2005 / 000300 filed on February 1, 2005. [0002] The present invention relates to a method for stimulating collagen synthesis and / or KGF expression, more specifically, a method for stimulating collagen synthesis and / or KGF expression using AIMP1 protein or a fragment thereof. Background technique [0003] The human body is covered by skin. Therefore, the skin is considered the largest organ of the human body and is generally composed of 2 layers, the epidermis and the dermis. The epidermis is the thinnest layer located on the outermost part of the skin, and plays an important role in moisturizing and protecting the skin. In detail, the epidermis prevents the skin from drying out, and also, prevents the intrusion of harmful substances such as ultraviolet rays, viruses, and bacteria. Furthermore, it naturally emulsifies the fat of the sebaceous glands and the water of the sweat gland...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/64
CPCA61K8/64A61Q19/08A61P17/00A61P17/02A61P17/16A61P43/00A61K8/65C07K14/00
Inventor 金圣勋
Owner 伊玛吉恩有限公司
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