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Nur77 phosphorylated derived peptide and application thereof in preparation of medicine for promoting embryo implantation

A technology of embryo implantation and phosphorylation, applied in the field of biomedicine, can solve undetermined problems, achieve the effect of promoting cell adhesion, good application potential, and promoting embryo implantation

Active Publication Date: 2022-05-03
NANJING DRUM TOWER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Phosphorylation of Nur77 is found in multiple cell types and phosphorylation regulates Nur77 function, but it has not been determined whether Nur77 expression or phosphorylation status in endometrial epithelial cells has an effect on epithelial-embryonic adhesion

Method used

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  • Nur77 phosphorylated derived peptide and application thereof in preparation of medicine for promoting embryo implantation
  • Nur77 phosphorylated derived peptide and application thereof in preparation of medicine for promoting embryo implantation
  • Nur77 phosphorylated derived peptide and application thereof in preparation of medicine for promoting embryo implantation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Preparation of Nur77 Phosphorylated Derivative Peptides and Non-Phosphorylated Derivative Peptides

[0019] The 361st to 371st amino acids of Nur77 (Nur77 short peptide) were artificially synthesized in vitro by the solid-phase peptide synthesis method in the chemical synthesis method. Its sequence is shown in SEQ ID NO.2, and the derivative peptide of Nur77 short peptide is Phosphorylated threonine at position 366 is obtained. In order to promote the entry of the short peptide into the cell to play a biological role, an amino acid (sequence shown in SEQ ID NO.2) with the function of penetrating the cell membrane is fused to the amino terminal of the Nur77 short peptide, and finally the Nur77 phosphorylated derivative peptide ( Phosphorylated peptide for short) and Nur77 non-phosphorylated derivative peptide (non-phosphorylated peptide for short) sequences are as follows figure 1 shown.

Embodiment 2

[0020] Example 2: The Nur77 phosphorylated derivative peptide of Example 1 promotes the adhesion of Ishikawa cells to BeWo cell spheres in vitro

[0021] The trophoblast cell line BeWo cell suspension culture is used to form BeWo cell spheres with a diameter of 150-200um. The endometrial epithelial cell line Ishikawa cells were adhered to form a monolayer of cells, and different concentrations of Nur77 phosphorylated peptides and non-phosphorylated peptides were added to the cell culture medium to incubate for 24 hours, and 100 BeWo cell spheres were mixed with the The monolayer Ishikawa cells in each group after treatment with the derived peptide were co-cultured for 1.5 hours, the medium was changed and the unadhered BeWo cell spheres were discarded, and the number of adhered BeWo cells was counted under a microscope.

[0022] The results are shown in Table 1. Compared with the blank control group, the adhesion percentage of Ishikawa cells treated with phosphorylated peptide...

Embodiment 3

[0027] Example 3: Using an in vitro model of mouse embryo adhesion to evaluate the effect of the Nur77 phosphorylated derivative peptide of Example 1 on promoting embryo adhesion

[0028] The endometrium epithelial cell line Ishikawa cells were used to form a single layer of cells, and different concentrations of Nur77 phosphorylated peptides and non-phosphorylated peptides were added to the cell culture medium to incubate for 24 hours, and five mouse blastocysts were mixed with Monolayer Ishikawa cells were co-cultured for 24 hours, and the degree of adhesion between blastocysts and Ishikawa cells treated with different derived peptides was evaluated under a microscope (grade 1-4).

[0029] The results are shown in Table 2. Compared with the blank control group, the degree of adhesion between Ishikawa cells treated with phosphorylated peptides and mouse blastocysts was significantly increased (P<0.05), while the treatment with non-phosphorylated peptides at the same dose The ...

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Abstract

The invention discloses an Nur77 phosphorylated derived peptide and application thereof in preparation of a medicine for promoting embryo implantation. The sequence of the phosphorylated derived peptide is shown as SEQ ID NO.1. Through a mouse in-vitro embryo adhesion test, an in-vivo epithelium-embryo adhesion test for delaying implantation into a mouse model and correlational research on the molecular cell level, the Nur77 phosphorylated derived peptide can be used for preparing a medicine for promoting embryo implantation. It is proved that the Nur77 phosphorylated derived peptide can enhance the uterine epithelium-embryo adhesion effect by promoting the beta 3 integrin mediated cell adhesion effect, so that embryo implantation is promoted; the medicine prepared by using the phosphorylated derived peptide can improve repeated implantation failure (RIF) caused by unknown reasons of women.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a Nur77 phosphorylated derivative peptide and its application in the preparation of medicines for promoting embryo implantation. Background technique [0002] Successful embryo implantation into the maternal endometrium is a critical step in mammalian reproduction, as successful embryo implantation requires embryo attachment to the endometrial epithelium and subsequent invasion of the decidualized endometrium. In humans, the natural conception rate in one menstrual cycle is approximately 30%, and embryo implantation failure accounts for approximately 75% of pregnancy losses. In the treatment of human assisted reproductive technology (ART), the incidence of repeated implantation failure (RIF) also reaches 10%, so it is effective for promoting embryo implantation, preventing and treating unexplained repeated implantation failure (RIF). Drugs are in great demand [1] . [00...

Claims

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Application Information

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IPC IPC(8): C07K14/705A61K38/17A61P37/06
CPCC07K14/70567A61P37/06A61K38/00
Inventor 颜桂军孙海翔姜瑞伟王海滨
Owner NANJING DRUM TOWER HOSPITAL
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