Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of antimicrobial peptide Protegrin-1 for preventing and controlling porcine reproductive and respiratory syndrome

A technology of porcine blue ear disease and antibacterial peptides, which is applied in the field of medicine, can solve the problems of short immune period, incomplete inactivation, scattered toxins, etc., and achieve the effect of good antiviral effect.

Inactive Publication Date: 2014-12-31
SUN YAT SEN UNIV
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(5) Mixed infection. At present, clinical mixed infection, especially the mixed infection of circovirus, Haemophilus parasuis and PRRSV, makes the prevention and control of PRRSV even more difficult.
Among them, the inactivated vaccine has the following disadvantages: (1) It requires large doses of vaccination or the application of concentrated antigens, and the immunity period is short, often requiring booster vaccination; (2) It cannot cause local immunity, so that the effect of cellular immunity is weak; (3) It produces complete immunity It takes 2-3 weeks, which is not conducive to emergency vaccination and the reduction of vaccine costs; (4) There is a possibility of incomplete inactivation and loose virus
Whether antimicrobial peptide PG-1 has antiviral effect on PRRSV virus has not been reported so far

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 antimicrobial peptide Protegrin-1 for preventing and controlling porcine reproductive and respiratory syndrome
  • Application of antimicrobial peptide Protegrin-1 for preventing and controlling porcine reproductive and respiratory syndrome
  • Application of antimicrobial peptide Protegrin-1 for preventing and controlling porcine reproductive and respiratory syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 PG-1 chemical synthesis and anti-Escherichia coli activity experiment

[0037] 1. First search the amino acid sequence of porcine antimicrobial peptide PG-1 through the antimicrobial peptide database (APD) The Antimicrobial Peptide Database (http: / / aps.unmc.edu / AP / main.php), as shown in SEQ ID NO.1 showed that the amino acid sequence was sent to Shanghai Gilbetter Biochemical Co., Ltd. (http: / / glbetter.cn.1688.com / ) for synthesis, because the β-sheet structure of PG-1 is crucial to its biological activity, and β- The formation of the folding structure depends on the 2 pairs of disulfide bonds in PG-1, so when synthesizing PG-1, it is necessary to ensure the successful synthesis and accurate positioning of the disulfide bonds. The two pairs of disulfide bonds are the 6th and 15th Cys, and the 8th and 13th Cys, respectively, and in order to enhance the biological activity of PG-1, its C-terminus was modified by amidation. HPLC 95% purity, synthesized 30mg. A...

Embodiment 2

[0042] Example 2 PG-1 Cytotoxicity Test

[0043] AlamarBlue (purchased from Invitrogen) is used as an indicator of living cell metabolism. Under the mitochondrial enzymatic reduction reaction, a measurable fluorescent metabolite will be produced, and the cell activity can be monitored by measuring its fluorescence intensity. Cultivate Marc-145 cells with DMEM medium containing 10% fetal bovine serum to 60-70%, discard the medium, add nutrient solution containing PG-1 times dilution for 36 hours, set PBS control group, and then add 10 % (V / V) ratio AlamarBlue continued to incubate for 3 hours, read the fluorescence values ​​of 540nm excitation light and 590nm emission light respectively with a multi-functional microplate reader, and make PG-1 cytotoxicity graph (see attached figure 2 ). Taking the cell activity of the PBS control group as 100%, the fluorescence value of the cells treated with PG-1 in multiple dilutions is compared with the fluorescence value of the PBS cont...

Embodiment 3

[0044] Embodiment 3 PG-1 antiviral test

[0045] 1. Cultivate Marc-145 cells in a 6-well plate containing DMEM medium containing 10% fetal bovine serum until the cell confluence reaches 70%, discard the culture medium, wash 3 times with PBS, and inoculate HP-PRRSV with MOI=0.1 , continue to culture at 37°C for 5h in DMEM medium with 2% fetal bovine serum.

[0046] 2. Wash 3 times with PBS, add 2% fetal bovine serum DMEM medium with a concentration of 20, 30, 40 mg / L PG-1 to the cells and incubate at 37°C for 36 hours, set up a PBS control group, wash 3 times with PBS, collect Supernatant for TCID 50 test, see attached image 3 ,Depend on image 3 It can be seen that PG-1 can significantly reduce the virus production in the cell supernatant and inhibit the release of the virus into the cell supernatant; in addition, add 400 μL Trizol to each well to extract RNA and perform qRT-PCR detection, see attached Figure 4 ,Depend on Figure 4 It can be seen that at the transcrip...

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 belongs to the technical field of medicines, and in particular discloses application of an antimicrobial peptide Protegrin-1 for preventing and controlling porcine reproductive and respiratory syndrome. According to the invention, an antimicrobial peptide Protegrin-1 is chemically synthesized according to an amino acid sequence of reported Protegrin-1 firstly; then, antimicrobial activity of the antimicrobial peptide Protegrin-1 is proved through an escherichia coli resistant DH5 alpha experiment, and researches on the antivirus activity of the antimicrobial peptide to HP-PRRSV are made through qRT-PCR, Western-Blot, cell supernatant TCID50 detection, IFA and other methods; and further the antivirus mechanism of the antimicrobial peptide is explained in virus adsorption and entrance into cell processes. The antimicrobial peptide Protegrin-1 is expected to be used as a novel medicine for preventing and controlling porcine reproductive and respiratory syndrome.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of antibacterial peptide Protegrin-1 in preventing and treating pig blue-ear disease. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS), also known as porcine blue ear disease, is caused by porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV). The disease first broke out in the United States in 1987, and then spread to Europe. The virus was isolated in my country in 1996. Currently, PRRSV is divided into two genotypes based on genome sequence and antigenicity differences, one is the European type represented by the Lelystad Virus (LV) strain, and the other is the American type represented by the ATCC VR-2332 strain . In my country, the American type of PRRSV is the main type, but it is reported that European type strains have also been isolated. In 2006, pig hypertherm...

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 Patents(China)
IPC IPC(8): A61K38/10A61K9/19A61K9/20A61K9/48A61K9/16A61P31/14
CPCY02A50/30
Inventor 刘小红郭春和莫德林陈瑶生高进涛
Owner SUN YAT SEN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com