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A human skin tissue endogenous polypeptide pdhps1 and its application

An endogenous, dermal technology, applied in applications, skin diseases, tissue culture, etc., can solve problems such as pain, localized adverse complications, and affecting the quality of life of patients

Active Publication Date: 2019-03-08
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinically, hypertrophic scars often cause local itching, pain, possible appearance damage and functional limitation, which bring great harm to the patient's body and mind, and seriously affect the quality of life of the patient.
At present, although surgical resection, local hormone injection, and laser intervention are important methods for the treatment of hypertrophic scars, they all have limitations or adverse complications, and there is still a lack of effective prevention and treatment methods

Method used

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  • A human skin tissue endogenous polypeptide pdhps1 and its application
  • A human skin tissue endogenous polypeptide pdhps1 and its application
  • A human skin tissue endogenous polypeptide pdhps1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Commissioned Shanghai Keyept Biotechnology Co., Ltd. to synthesize the polypeptide PDHPS1 shown in SEQ ID NO.1 as follows by solid-phase method:

[0029] Ile Ala Thr Thr Thr Thr Ala Ser Ala Ala Thr Ala Ala Ala Ile Gly Ala ThrPro Arg Ala Lys

[0030] This is a polypeptide composed of 21 amino acids. The main biological parameters of the aforementioned polypeptide sequence obtained through online tools are as follows:

[0031] The isoelectric point (PI) is 11.0, and the molecular mass (Mw) is 1915.18Da.

[0032] Take 10mg of polypeptide, dilute it with sterile double distilled water to 50mM-40°C and store it for later use.

Embodiment 2

[0033] Example 2 CCK-8 method to detect the effect of polypeptide PDHPS1 on the proliferation of human scar dermal fibroblasts

[0034] At 37°C, 5% CO 2 In the incubator, human scar dermal fibroblasts were cultured, seeded in a 96-well culture plate at a density of 2,000 cells / well, and 100 μl of medium / well was added. After 24 hours, the polypeptide PDHPS1 was added for treatment (final concentrations were 0.1 , 1, 10, 100 μM), the control group was treated with sterile double-distilled water, and each group had 5 replicate wells, cultured in a 37°C incubator.

[0035] After 24 hours, take out the 96-well plate, add 10 μl CCK-8 to each well to avoid the formation of air bubbles, and return to the 37-degree incubator to continue incubation for 2 hours. The absorbance was measured by a microplate reader, counted at a wavelength of 450nm, and the growth curve was drawn.

[0036] The result is as figure 1 As shown, it can be seen that the polypeptide PDHPS1 has the function of...

Embodiment 3

[0037] Example 3 Real-time quantitative PCR (RT-PCR) detection of the effect of polypeptide PDHPS1 on the mRNA expression of collagen genes COL1A1 and COL1A2 in human scar dermal fibroblasts

[0038] At 37°C, 5% CO 2 In the incubator, cultivate human scar dermal fibroblasts, press 10 5 Seed in a 6-well plate at a density of / ml. When the cell confluence reaches about 60%, add the polypeptide PDHPS1 (final concentration 100 μM) into the human scar dermal fibroblasts, and use the RNA extraction kit (TRIzol method) to extract the cells after 24 hours total RNA.

[0039] The human scar dermal fibroblasts in the polypeptide treatment group and the control group were collected respectively, and the total RNA was extracted using the RNA extraction kit (TRIzol method). Primers were used to detect the expression of human COL1A1 and COL1A2. The Realtime PCR reaction system is: 10 μl of 2XSybr Mix, 1 μl of cDNA, 1 μl of primers, 8 μl of double distilled water, a total of 20 μl, run on...

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PUM

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Abstract

The invention discloses a human skin tissue endogenous polypeptide PDHPS1 and an application thereof. The amino acid sequence of the human skin tissue endogenous polypeptide PDHPS1 is shown in SEQ IDNO.1. The polypeptide PDHPS1 inhibits the propagation of a scar dermal fibroblast, inhibits the expression of mRNA of collagen genes COL1A1 and COL1A2 in the scar dermal fibroblast, and obviously reduces the protein expression of a main marker alpha-SMA of a myofibroblast in the scar dermal fibroblast. The human skin tissue endogenous polypeptide PDHPS1 can be used to prepare a drug or agent whichcan inhibit the propagation of the scar dermal fibroblast, reduce the collagen content and reduce the amount of the myofibroblast, and provides a new target spot for inhibiting the scar hypertrophy.

Description

technical field [0001] The invention relates to the fields of wound healing and scars, in particular to an endogenous polypeptide PDHPS1 of human skin tissue and its application. Background technique [0002] Hypertrophic scar is a common pathological scar, which is the result of excessive tissue repair and excessive deposition of extracellular matrix such as collagen in the lesion during the healing process after the damaged dermis of the skin, and its incidence rate is as high as 70%. Clinically, hypertrophic scars often cause local itching, pain, possible appearance damage and functional limitation, which bring great harm to the patient's body and mind, and seriously affect the quality of life of the patient. At present, although surgical resection, local hormone injection, and laser intervention are important methods for the treatment of hypertrophic scars, they all have limitations or adverse complications, and there is still a lack of effective prevention and treatment...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C12N15/12C12N5/00A61K38/17A61P17/00
CPCA61K38/00A61P17/00C07K14/47C12N5/0018C12N2501/999
Inventor 李俊李景云季晨博曾新崔县伟付子毅
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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