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Application of glycyrrhizin in resistance to PEDV (porcine epidemic diarrhea virus) infection

A porcine epidemic diarrhea and virus infection technology, applied in the direction of antiviral agents, medical preparations containing active ingredients, organic active ingredients, etc., to achieve the effect of low drug residues

Inactive Publication Date: 2017-02-22
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has not been reported that liquiritigenin inhibits the infection of PEDV

Method used

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  • Application of glycyrrhizin in resistance to PEDV (porcine epidemic diarrhea virus) infection
  • Application of glycyrrhizin in resistance to PEDV (porcine epidemic diarrhea virus) infection
  • Application of glycyrrhizin in resistance to PEDV (porcine epidemic diarrhea virus) infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The assay result of the cytotoxicity of embodiment 1 Liquiritigenin on Vero cells

[0037] After Vero cells were counted, they were diluted with DMEM nutrient solution containing 10% fetal bovine serum to an appropriate density at 2×10 3 The concentration per well was added dropwise to a 96-well plate and placed at 37°C, 5% CO 2 After the cells adhere to the wall to form a monolayer (about 17-18h) in the incubator, dilute the liquiritigenin with DMEM nutrient solution: 0.1mM, 0.2mM, 0.4mM and 0.8mM, and do six repetitions for each concentration, at 96 After treating the Vero cells with 100 μl / well of the diluted glycyrrhizin sample on the well cell culture plate for 24 hours, take 50 μl of the supernatant, centrifuge for 5 minutes (250 × g), and then transfer to the microplate well, and then use CytoTox Non-RadioactiveCytotoxicity Assay kit (promega company) detects, at OD490nm, read data ( figure 1 ),Depend on figure 1 It can be seen that the amount of LDH (lactate...

Embodiment 2

[0038]Example 2 Liquiritigenin inhibits the activity of porcine epidemic diarrhea virus infection on Vero cells (Western blot and fluorescent quantitative PCR, plaque formation test)

[0039] (1) Western blot assay for the activity of liquiritigenin in inhibiting porcine epidemic diarrhea virus infection on Vero cells

[0040] After Vero cells were counted, they were diluted with DMEM nutrient solution containing 10% calf serum to an appropriate density at 5×10 5 The concentration per well was added dropwise to a 6-well plate and placed at 37°C, 5% CO 2 Wait for the cells to adhere to the wall and form a monolayer in the incubator (about 17-18h). Liquiritigenin was diluted to a specific concentration with serum-free DMEM: 0.1mM-0.8mM, and after pre-incubating with Vero cells at 37°C for 2h, inoculated porcine epidemic diarrhea virus HLJBY strain (MOI=0.1) in Vero cells, 37°C, 5 %CO 2 Incubate in the incubator for 1 hour with the presence of glycyrrhizin, wash three times wi...

Embodiment 3

[0050] The influence of embodiment 3 liquiritigenin on porcine epidemic diarrhea virus infection process (Western blot, fluorescent quantitative PCR and plaque formation test)

[0051] (1) Effect of liquiritigenin on cell entry and replication of porcine epidemic diarrhea virus (Western blot)

[0052] After Vero cells were counted, they were diluted with DMEM nutrient solution containing 10% calf serum to an appropriate density at 5×10 5 The concentration per well was added dropwise to a 6-well plate and placed at 37°C, 5% CO 2 Wait for the cells to adhere to the wall and form a monolayer in the incubator (about 17-18h). Liquiritigenin was diluted to a specific concentration with serum-free DMEM: 0.1mM-0.8mM, and after pre-incubating with Vero cells at 37°C for 2h, inoculated porcine epidemic diarrhea virus HLJBY strain (MOI=0.1) in Vero cells, 37°C, 5 %CO 2 Incubate in an incubator for 1 hour with the presence of liquiritigenin, wash three times with PBS, add 2% calf serum...

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Abstract

The invention mainly relates to activity of a drug glycyrrhizin with anti-PEDV (porcine epidemic diarrhea virus) activity in resistance to host cells infected with a PEDV HLJBY strain, and belongs to the field of antivirus applications. The drug glycyrrhizin is an extract of traditional Chinese medicine liquorice and can effectively inhibit endocytosis and duplication of PEDV, thereby effectively inhibiting PEDV infection. Vero cells verify that glycyrrhizin performs the anti-virus effect mainly by inhibiting virus endocytosis and duplication.

Description

[0001] 1. Technical field [0002] The invention belongs to the field of antiviral applications, and relates to the application of anti-porcine epidemic diarrhea virus drug liquiritigenin. [0003] 2. Background technology [0004] Porcine epidemic diarrhea (porcine epidemic diarrhea, PED) is caused by porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV), an infectious disease of pigs characterized by watery diarrhea, vomiting, and dehydration. The disease often breaks out into a pandemic in winter, and pigs of all ages can be infected with PEDV, but suckling piglets are the most susceptible, with a morbidity rate of up to 100% and a mortality rate of 80%-100%. Clinically, it is co-infected with porcine transmissible gastroenteritis and porcine rotavirus diarrhea. Although PED first occurred in Europe, it has become a serious infectious disease in pig farming in Asian countries including China, Japan, Korea, Thailand and the Philippines, causing huge economi...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P1/12A61P31/14
CPCA61K31/352
Inventor 郇长超茅翔钱莺娟
Owner NANJING AGRICULTURAL UNIVERSITY
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