Application of Chinese softshell turtle Cathelicidin-Ps-CATH4 peptide in preparation of anti-inflammatory drug

A technology of ps-cath4 and Chinese soft-shelled turtle, which is applied in the field of biomedicine, can solve the problems of no reports, etc., and achieve the effect of simple structure, good stability, convenient chemical synthesis and genetic engineering preparation

Active Publication Date: 2015-06-24
DALIAN UNIV OF TECH
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese soft-shelled turtle belongs to the genus Sino soft-shelled turtle of the family Turtleidae of the order Turtle, and there is no report on the research and application of the cathelicidin host defense peptide of the Chinese soft-shelled turtle.

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 Chinese softshell turtle Cathelicidin-Ps-CATH4 peptide in preparation of anti-inflammatory drug
  • Application of Chinese softshell turtle Cathelicidin-Ps-CATH4 peptide in preparation of anti-inflammatory drug
  • Application of Chinese softshell turtle Cathelicidin-Ps-CATH4 peptide in preparation of anti-inflammatory drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Ps-CATH4 Peptide Inhibits LPS-Induced Expression of Inflammatory Cytokines and NO Synthase (INOS) Gene

[0058] Mouse macrophage RAW264.7 was cultured in DMEM medium containing 10% fetal bovine serum at 37°C CO 2 Culture in an incubator. When the cells grow to the logarithmic growth phase in good condition, digest with 0.25% trypsin, discard the trypsin, and blow down the adherent cells with DMEM medium with 10% fetal bovine serum. After pipetting evenly, count the cells and adjust the cell concentration to 1x10 6 / ml. Add the cells evenly pipetted into a 24-well plate, 1ml per well, in 37°C CO 2 Cultivate overnight in the incubator to allow the cells to adhere to the wall, then suck out the supernatant and discard it, add 960ul of fresh DMEM medium containing 10% fetal bovine serum, add 20ul of LPS at a certain concentration to the positive control group and the drug-dosed group, and then add 20ul was diluted with DMEM and incubated with different con...

Embodiment 2

[0060] Example 2 Ps-CATH4 peptide regulates LPS-induced secretion of inflammatory cytokines and NO in vitro

[0061] Mouse macrophages RAW264.7 were cultured in DMEM medium containing 10% fetal bovine serum to logarithmic phase of growth, digested with trypsin, and seeded in 96-well plates, 1x10 per well 6 indivual. CO at 37 °C 2 Cells were cultured overnight in an incubator. After the cells adhere to the wall, discard the supernatant, add 180ul of fresh DMEM medium containing 10% fetal bovine serum to each well, then add 10ul of different concentrations of Ps-CATH4 and 10ul of a certain concentration of LPS to each well, and add the blank 20ul serum-free DMEM medium, treated for 24h. The samples treated with drugs for 24 hours were used to collect the supernatant to detect the release of NO and inflammatory factors with Griess kit and ELISA kit, respectively, and 5 replicates were set for each experimental group. The Griess kit was purchased from Biyuntian Bioreagent Co...

Embodiment 3

[0062] Example 3 Western blot method to detect the effect of Ps-CATH4 on MAPK inflammatory signaling pathway

[0063] Mouse macrophages RAW264.7 were cultured in DMEM medium containing 10% fetal bovine serum to the logarithmic phase of growth, digested with trypsin, and mixed with 3x10 6 cells / ml inoculated in 6-well plate, 2ml per well. CO at 37°C 2 Cultivate overnight in the incubator to allow the cells to adhere to the wall, then discard the supernatant culture medium of the cells and wash with phosphate buffered saline (PBS: 0.8g NaCl; 0.2g KCl; 2.9g NaCl 2 HPO 4 .12H 2 O; 0.27g KH 2 PO 4 Dissolve in 800ml deionized water, stir to dissolve, dilute to 1L, adjust pH to 7.4 with concentrated HCl, and autoclave. ) Wash the cells 2-3 times, and replace with serum-free DMEM medium for starvation treatment for 16 hours. Add 20ul of samples of different concentrations to each well (to make the final concentration of each well 5, 10, 20ug / ml), add 20ul of serum-free DMEM...

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 discloses application of Chinese softshell turtle Cathelicidin-Ps-CATH4 peptide in preparation of an anti-inflammatory drug, and belongs to the technical field of biomedicine. Ps-CATH4 is linear peptide, and contains 32 amino acid residues; RT-PCR displays that the peptide has an inhibition effect on expression of critical inflammatory factors induced by lipopolysaccharide and an inducible NO synthase gene; and on the protein level, the peptide has a negative regulation effect on LPS-induced inflammatory factors and NO release, has an ascending effect on expression of inflammatory factors, and is mainly used for inhibiting release of the inflammatory factors by inhibiting activation of a TLR4 acceptor and a downstream MAPK signal channel. The peptide is simple in structure, does not contain a disulfide bond or a cyclic structure; chemical synthesis and genetic engineering preparation are facilitated; the primary medicinal evaluation shows low cytotoxicity and hemolytic activity; and the peptide is good in stability on salt, blood serum, temperature and time, and is applied to preparation of the anti-inflammatory drug.

Description

technical field [0001] The invention provides a kind of soft-shelled soft-shelled turtle ( Pelodiscus sinensis ) cathelicidin-Ps-CATH4 and its encoding gene, amino acid sequence and application in the preparation of anti-inflammatory drugs, belonging to the field of biomedical technology. Background technique [0002] Cathelicidin is a family of multifunctional immunomodulatory peptides composed of an N-terminal signal peptide region, a middle conserved cathelin domain and a C-terminal highly specific mature peptide region. It is found in almost all kinds of animals (mammals, birds, Both amphibians and fish) have been found. Like defensin, Cathelicidin is a host defense peptide unique to most vertebrates including humans. It constitutes a key component of the natural immune response of vertebrates and is a link The bridge between natural immunity and specific immunity plays an extremely important role in the animal innate immune system. In addition to its broad-spectrum an...

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): C07K14/46C12N15/12A61K38/17A61P29/00
CPCA61K38/00C07K14/46
Inventor 于海宁王义鹏乔雪鲁一灵王晨
Owner DALIAN UNIV OF TECH
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