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Application of calcium lactate in preparation of medicine for preventing and treating infection caused by spring viremia of carp virus (SVCV)

A technology of carp spring viremia and virus infection, applied in the direction of antiviral agents, anhydride/acid/halide active ingredients, aluminum/calcium/magnesium active ingredients, etc., can solve the problems of immature aquatic animal virus diseases, etc. No drug residues, single ingredients, no toxic and side effects

Active Publication Date: 2020-04-21
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the prevention and treatment methods for viral diseases in aquatic animals are not mature enough, and the research on SVCV virus vaccines and therapeutic drugs is still in its infancy, and effective and rapid prevention and treatment methods are urgently needed to effectively control the outbreak of the virus

Method used

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  • Application of calcium lactate in preparation of medicine for preventing and treating infection caused by spring viremia of carp virus (SVCV)
  • Application of calcium lactate in preparation of medicine for preventing and treating infection caused by spring viremia of carp virus (SVCV)
  • Application of calcium lactate in preparation of medicine for preventing and treating infection caused by spring viremia of carp virus (SVCV)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Calcium lactate enhances the function of cells against SVCV infection

[0020] 1. Fish cells (EPC) were prepared at 1×10 per well 5 After seeding the number of cells into 24-well plate of cell culture plate and culturing overnight, add 1 μl SVCV (MOI=1) to each well, and add 40mM CaLa / CuCl respectively at the same time 2 / FeCl 2 / MgCl 2 / MnCl 2 / ZnCl 2 The cells were treated for 24 hours, and the cytopathic effect was observed under the microscope. The cells were fixed with PFA and stained with crystal violet. The proportion of dead cells was significantly reduced ( figure 1 A), which shows that calcium lactate treatment of cells can significantly enhance the antiviral ability of cells. In order to further confirm this result, after EPC cells were infected with SVCV, different concentrations of calcium lactate were added to treat the cells for 48 hours, and the occurrence of cytopathic effect was observed under a microscope ( figure 1 B), collect the s...

Embodiment 2

[0022] Example 2: Expression of apoptosis factor p53 after calcium lactate treatment of fish cells infected with SVCV

[0023] The previous research work of the inventors showed that SVCV-infected cells are closely related to the expression of p53. In order to detect the expression levels of endogenous and exogenous p53 after calcium lactate treatment, ZFL cells or EPC cells were seeded in 6-well plates and cultured for 24 hours. Wherein EPC cells were transfected with 2 μg pCMV-Myc or Myc-p53, then treated with different concentrations of calcium lactate (20, 40 mM) for 24 h, and the cells were collected to detect the expression of p53 protein in the cells by Western Blot ( image 3 A and 3B), endogenous p53 was detected with p53 antibody, and exogenous p53 was detected with Myc tag antibody. The results showed that after calcium lactate treatment, the expression of endogenous or exogenous p53 protein increased significantly and had a dose-dependent effect. In addition, in or...

Embodiment 3

[0024] Example 3: Calcium lactate treatment of ubiquitination modification of p53 protein in fish cells infected with SVCV

[0025] 1. As a reversible protein modification, ubiquitination is mainly involved in regulating the stability, activity and localization of target proteins. Calcium lactate treatment of cells can promote the expression of p53, so whether calcium lactate treatment affects the ubiquitination of p53 was detected. First inoculate EPC cells in 10cm 2 Cell culture dishes were cultured overnight and transfected with 5 μg Myc-p53 and 1 μg HA-Ub. After 24 hours, they were treated with 20 and 40 mM calcium lactate for 18 hours, and then treated with MG132 for 6 hours. Cells were collected for ubiquitination experiments and Western Blot analysis ( Figure 4 A). The results showed that calcium lactate treated cells could promote the ubiquitination of p53 in a dose-dependent manner.

[0026] 2. The previous research work of the inventors showed that SVCV-infecte...

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Abstract

The invention discloses an application of calcium lactate in preparation of a medicine for preventing and treating infection caused by spring viremia of carp virus (SVCV). An experimental animal infection model proves that calcium lactate can play a role in treating SVCV-caused infection of zebra fish, virus proliferation and qPCR experiments are combined to find that the expression quantity of virus-related genes after calcium lactate treatment is obviously reduced, so that calcium lactate can be used as a medicine for preventing and treating SVCV and has the advantages of low dosage, safetyand no toxic or side effect.

Description

technical field [0001] The invention belongs to the technical field of aquaculture, and in particular relates to the medical application of calcium lactate, which can be used for the preparation, prevention and treatment of aquatic animal diseases caused by carp spring viremia virus infection. Background technique [0002] Calcium is an essential element for living organisms. Whether in the muscles, nerves, body fluids or bones of organisms, there are 2+ bound protein. Calcium participates in metabolism and is an essential element in processes such as nerve transmission, muscle contraction, blood clotting, and hormone release. It is also the main component of bones, and its importance is evident. [0003] Calcium usually exists in the form of calcium ions in organisms, and participates in various biological processes in this form. Among them, in terms of resisting external stimuli, generally, foreign antigens activate T cell receptors, which will initiate calcium ion-media...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/19A61K33/06A61P31/14
CPCA61K31/19A61K33/06A61P31/14
Inventor 李顺陆龙凤
Owner INST OF AQUATIC LIFE ACAD SINICA
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