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Placenta-derived endogenous polypeptide and application thereof

An endogenous, placental technology, applied in the field of biomedicine, to achieve the effect of promoting trophoblast cell invasion, promoting cell proliferation, and promoting placental trophoblast cell invasion

Active Publication Date: 2022-03-18
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, the research progress of pravastatin, melatonin, nicotinamide, etc. in improving vascular endothelial cell injury has not obtained the exact clinical efficacy and practical application, but it has provided new clues for the treatment of preeclampsia

Method used

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  • Placenta-derived endogenous polypeptide and application thereof
  • Placenta-derived endogenous polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Screening of endogenous polypeptides derived from human placenta

[0027] The placentas of severe preeclampsia patients (3 cases) and healthy pregnant women (3 cases) were collected at 30-39 weeks of gestation and delivered by cesarean section, and the maternal surface tissue pieces (about 1 cm in diameter) were collected and washed with sterile phosphate-buffered saline Blood is removed and the tissue is homogenized, ultrafiltered and desalted to obtain peptides. Peptides with significant differences between the two groups of placentas were screened (P value ≤ 0.05), and Blast2go software was used for GO bioinformatics analysis, through the KEGG website (KEGG; http: / / www.genome.jp / kegg ) to conduct pathway analysis on the polypeptide precursor protein, and use the ExPASy online tool (https: / / web.expasy.org / compute.pi / ) to calculate the isoelectric point (pI) and molecular weight (MW) of the polypeptide. Selecting two groups of placenta with high background level and ...

Embodiment 2

[0029] Entrust Shanghai Keyept Biotechnology Co., Ltd. to synthesize the polypeptide shown in SEQ ID NO.1 by solid phase method: ArgAsn Asp Glu Glu Leu Asn Lys Leu Leu Gly Gly Val Thr Ile Ala Gln Gly Gly ValLeu Pro Asn Ile (SEQ ID NO. 1)

[0030] This is a polypeptide consisting of 27 amino acids. The main biological parameters of the aforementioned polypeptide sequence obtained through online tools are as follows: the isoelectric point (PI) is 4.68, and the molecular mass (Mw) is 2819.21Da.

[0031] Take 10mg of polypeptide, dilute it with sterile double distilled water to 50mM-40°C and store it for later use.

Embodiment 3

[0032] Example 3 CCK-8 method to detect the effect of polypeptide on the proliferation of human chorionic trophoblast cells (HTR-8 / Svneo)

[0033] In a 37°C, 5% CO2 incubator, human chorionic trophoblast cells were cultured, seeded in a 96-well culture plate at a density of 2000 cells / well, added 100 μl of medium / well, and treated with peptides (final concentration 0.5, 1.0, 5.0, 10.0 μM), the control group was treated with sterile double distilled water, and each group had 5 replicate wells, cultured in a 37°C incubator.

[0034] After 24 hours, 48 ​​hours, and 72 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.

[0035] The result is as figure 1 As shown, it can be seen that the polypeptide can obviously promote the proliferation o...

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Abstract

The invention discloses a placenta-derived endogenous polypeptide and application thereof. The invention relates to a placenta tissue endogenous polypeptide APEP1 (Anti-Pre-eclampsia peptide 1). The amino acid sequence of the placenta tissue endogenous polypeptide APEP1 (Anti-Pre-eclampsia peptide 1) is as shown in SEQ ID NO. 1. The polypeptide APEP1 can promote the proliferation capacity of placental trophoblasts. The human placenta tissue endogenous polypeptide APEP1 can be used for preparing drugs or reagents for promoting placenta trophoblast cell proliferation capacity of preeclampsia patients, and is expected to provide a novel treatment strategy for preeclampsia treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a placenta-derived endogenous polypeptide and its application. Background technique [0002] Although pre-eclampsia (PE) is an idiopathic disease during pregnancy, it can cause important organ damage (hypertensive encephalopathy, renal function damage, heart failure), placental abruption, premature delivery, and fetal growth impairment in pregnant women. Complications such as limitation are harmful. The global incidence of PE is about 4.6%, causing about 70,000 maternal deaths every year, while the incidence in some areas of my country is even as high as 10.49%. The current clinical treatment of PE is mainly to reduce blood pressure and prevent convulsions, but the effect is limited. Termination of pregnancy is often the last resort, so it is often accompanied by iatrogenic premature birth. In addition, the risk of long-term adverse cardiovascular events in PE mothers and their offsprin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N5/073A61K38/17A61P15/00
CPCC07K14/47A61P15/00C12N5/0605C12N2501/998A61K38/00
Inventor 龙伟李春艳吕明明尤梁惠李景云夏卿张艳榕
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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