Application of soybean peptide in promoting osteogenic activity

A technology of soybean peptide and osteoblasts, which is applied in the direction of cell culture active agents, bone/connective tissue cells, and microbial measurement/testing, and can solve problems such as unclear osteogenesis mechanisms

Pending Publication Date: 2022-05-13
YANTAI UNIV
View PDF4 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In previous studies, our research group has found that fermented yogurt has osteogenic activity in in vitro osteoblast experiments, and used ultrafiltration and metal ion chelation affinity chromatography to separate and purify peptides with the ability to bind calcium ions , its amino acid composition was identified as DEDEQIPSHPPR by LC-MS / MS, but its osteogenesis mechanism is still unclear. Therefore, this project aims to evaluate the formation of soybean peptides in zebrafish larvae and preosteoblast MC3T3-E1 Bone activity and osteogenesis mechanism, providing scientific basis and theoretical basis for the application of soybean peptide in functional foods as a potential osteogenic ingredient

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of soybean peptide in promoting osteogenic activity
  • Application of soybean peptide in promoting osteogenic activity
  • Application of soybean peptide in promoting osteogenic activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Effect of Soybean Peptide CBP on Osteoblast Proliferation

[0025] In this study, MC3T3-E1 was 2 × 10 by MTT method 4 / cm 2 The density was inoculated in a 96-well plate, and the proliferation of 1, 10, 100 μg / mL soybean peptide and osteoblasts were measured for 24, 48, and 72 hours of co-incubation to analyze whether soybean peptide had a proliferation effect on osteoblasts.

[0026] see results figure 1 As shown, it can be seen that soybean peptide CBP can promote the proliferation of osteoblasts in a dose- and time-dependent manner.

Embodiment 2

[0027] Example 2 Effect of Soybean Peptide CBP on Osteoblast Differentiation

[0028] The differentiation of osteoblasts is crucial to the anabolic metabolism of bone. During the differentiation of osteoblasts, alkaline phosphatase (ALP) and osteocalcin (OCN) are characteristic phenotypic markers in the early and late stages of differentiation, respectively. , we used ELISA kits to measure the activity of alkaline phosphatase and the secretion of osteocalcin during the differentiation process.

[0029] 1) Effect of soybean peptide CBP on alkaline phosphatase ALP

[0030] MC3T3-E1 cells were 2 × 10 4 / cm 2 Inoculate in a 24-well plate at a density of 24 wells, culture in complete medium for 2 days, change to differentiation medium and continue to culture for 4 days, change to serum-free medium on the 7th day, and add 100 μg / mL soybean peptide to co-incubate with osteoblasts After 72 hours, the cells were broken, and the protein was extracted by centrifugation. The protein co...

Embodiment 3

[0035] Example 3 Effect of Soybean Peptide on Osteoblast Mineralization

[0036] Osteoblast differentiation was induced by differentiation medium for 35 days, during which the differentiation medium was replaced every two days, 100 μg / mL soybean peptide was added at the same time during the differentiation, and osteoblasts were specifically stained by alizarin staining solution, under the microscope The staining was observed and photographed, and the effect of soybean peptide on the mineralization of osteoblasts was analyzed according to the staining area. The specific steps are:

[0037] The osteoblasts were stained with Alizarin-S stain to measure the level of matrix mineralization and comprehensively measure the differentiation level of osteoblasts. Cells were treated with different concentrations (0.7, 7, and 70 μM) of CBP for different days (7, 14, 21, and 28 days) in a medium containing ascorbic acid and β-glycerophosphate, and then stained with Alizarin Red , and capt...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides an application of a soybean peptide in promoting osteogenic activity, which comprises the following steps: detecting the proliferation level of osteoblasts, the activity of a marker ALP in the early stage of differentiation and a marker OCN in the later stage of differentiation, and detecting that the soybean peptide CBP promotes the proliferation and differentiation of the osteoblasts on the biochemical level; in the process of promoting osteoblast differentiation by the soybean peptide CBP, three classical signal pathways, namely ERK, JNK and p38 bypasses, of an MAPK signal pathway participate in the process and a key signal transduction step; the soybean peptide CBP promotes gene expression and phosphorylation level of a key factor Runx2 of an MAPK signal channel in osteoblasts.

Description

technical field [0001] The invention relates to the application of soybean peptide in promoting bone activity and belongs to the field of biomedicine. Background technique [0002] As a natural active protein derived from soybean, soy protein has always been the focus of attention of researchers in the field of nutrition. Numerous literatures have reported the functional activities of soybean protein, such as: regulating immunity and promoting the balance of intestinal flora , anti-oxidation, anti-cancer effects, etc. However, the research on small molecule soybean peptides is relatively lagging behind, which limits the further development and application of soybeans. Therefore, it is urgent to elucidate the osteogenesis mechanism of soybean peptides from the cellular and molecular levels. [0003] In terms of in vitro experiments, Kyungho et al. reported in 2014 that soybean extracts can indirectly inhibit osteoclast differentiation by regulating osteoclast factors produce...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12Q1/02G01N33/68
CPCC12N5/0654G01N33/5005G01N33/68C12N2501/998
Inventor 甘晶程永强王郐田王振华于天英孔潇李子家
Owner YANTAI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap