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A kind of human adipose stem cell secreted polypeptide adscp2 and its application

A technology of human adipose stem cells and composition, applied in the application, animal cells, vertebrate cells, etc., to achieve the effect of reducing the amount of myofibroblasts and reducing the content of collagen

Active Publication Date: 2022-04-01
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are currently no peptide drugs for the treatment of hypertrophic scars

Method used

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  • A kind of human adipose stem cell secreted polypeptide adscp2 and its application
  • A kind of human adipose stem cell secreted polypeptide adscp2 and its application
  • A kind of human adipose stem cell secreted polypeptide adscp2 and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Entrust Shanghai Keyept Biotechnology Co., Ltd. to synthesize the following sequence shown in SEQ ID NO.1 by solid-phase method:

[0022] Asp Glu Asn Arg Glu Lys Val Asn Asp Gln Ala Lys Leu

[0023] This is a polypeptide composed of thirteen amino acids. The main biological parameters of the aforementioned polypeptide sequence obtained through online tools are as follows:

[0024] The isoelectric point (PI) is 4.78, and the molecular mass (Mw) is 1558.67Da.

[0025] The above sequence is a natural sequence derived from the culture supernatant of human adipose stem cells. In addition to obtaining by chemical solid-phase synthesis, it can also be obtained by enzymatic digestion of long-chain polypeptides, and can also be cloned and expressed by host microorganisms or cells. The DNA fragment carrying the nucleotide sequence encoding one of the polypeptides is prepared by existing recombinant DNA technology. The nucleotide sequence encoding the above polypeptide is: gatga...

Embodiment 2

[0028] 1. Test materials

[0029] The RNA reverse transcription kit and RNA detection kit were purchased from Nanjing Novozyme Company, and the fibroblast culture medium was purchased from Sciencell Company of the United States.

[0030] 2. Synthesis and dilution of ADSCP2 polypeptide

[0031] The polypeptide ADSCP2 obtained by solid-phase synthesis in Example 1 of this experiment is taken as an example. Take 10mg of polypeptide, dilute it with sterile double distilled water to 50mM-40°C and store it for later use.

[0032] 3. Real-time quantitative PCR (RT-PCR) detection of the effect of polypeptide ADSCP2 on the mRNA expression of collagen genes COL1A1, COL1A2, COL3A1 in human scar dermal fibroblasts and the main marker of myofibroblasts ACTA2

[0033] At 37°C, 5% CO 2 In the incubator, cultivate human scar dermal fibroblasts, press 10 5 Seed in 6-well plates at a density of 1 / mL, and when the cell confluency reaches about 60%, add polypeptides (final concentrations of 1...

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Abstract

The invention discloses a human adipose stem cell secreted polypeptide ADSCP2 and its application. A human adipose stem cell secreted polypeptide ADSCP2, the amino acid sequence of which is shown in SEQ ID NO.1. The polypeptide ADSCP2 can inhibit the mRNA expression of collagen genes COL1A1, COL1A2, and COL3A1 in human scar dermal fibroblasts, and significantly reduce the mRNA expression of the main myofibroblast marker ACTA2 in human scar dermal fibroblasts. The human adipose stem cell secreted polypeptide ADSCP2 can be used to prepare drugs or reagents for reducing the collagen content of fibroblasts in scar dermis and the amount of myofibroblasts, providing a new target for inhibiting scar hyperplasia.

Description

technical field [0001] The invention relates to the fields of wound healing and scars, in particular to a human adipose stem cell secreted polypeptide ADSCP2 and its application. Background technique [0002] Hypertrophic scar is a kind of skin fibrotic disease, which is mainly characterized by local excessive deposition of extracellular matrix such as collagen, and it is a huge challenge for burns and plastic surgery. It not only affects the appearance, but also causes symptoms such as itching, pain, contracture and even disability, seriously reduces the quality of life of patients, and brings great pain to patients' body and mind. The incidence rate of hypertrophic scar is 39%-68% in patients with skin injury and surgery, and the incidence rate in burn patients is as high as 91%. At present, local pressure, laser intervention, drug injection, surgical resection and other methods are mostly used to treat hypertrophic scars, but all of them have limitations and adverse comp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C12N15/12C12N5/0775C12N15/85A61K38/17A61P17/02
CPCC07K14/47C12N5/0667A61P17/02A61K38/00
Inventor 李景云马洁桦文娟龙伟祝丽珺
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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