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Application of Metformin in inhibiting bovine viral diarrhea virus infection

A technology for bovine viral diarrhea and virus infection, applied in the direction of antiviral agents, organic active ingredients, medical preparations containing active ingredients, etc.

Active Publication Date: 2021-06-25
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether Metformin affects BVDV-infected host cells has not been reported yet.

Method used

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  • Application of Metformin in inhibiting bovine viral diarrhea virus infection
  • Application of Metformin in inhibiting bovine viral diarrhea virus infection
  • Application of Metformin in inhibiting bovine viral diarrhea virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment one: the purposes of Metformin in suppressing bovine viral diarrhea virus infection

[0035] The first aspect of the present invention provides the application of Metformin in the preparation of drugs for preventing and / or treating bovine viral diarrhea virus infection.

[0036] According to the present invention, the concept of "prevention and / or treatment" refers to any measure suitable for the treatment of bovine viral diarrhea virus-related diseases, or for the prophylactic treatment of this manifested disease or the manifested symptoms, or to avoid such Relapse of a disease, such as relapse after the end of the treatment period or treatment of symptoms of a disease that has already occurred, or pre-intervention to prevent or inhibit or reduce the occurrence of such diseases or symptoms.

[0037] At the same time, in the drug for preventing and / or treating bovine viral diarrhea virus infection, the drug concentration of Metformin is not lower than 10 mM. ...

Embodiment 2

[0047] Example 2: MTT detects the effect of Metformin on the proliferation of MDBK cells

[0048] The MDBK cells grown to the logarithmic phase were digested and collected, and 5000 cells per well were passaged to a 96-well plate, and 100 μL of cell culture medium (90% DMEM + 10% FBS + 1% double antibody) was added to each well, and placed in the cell After culturing in the incubator for 24 hours, add 0, 5, 8, 10, 15, 20 and 50 mM Metformin for 0, 12, 24, 36 and 48 hours respectively; discard the supernatant carefully at the end of the treatment, and add 100 μL to each well Cell culture medium and 10 μL MTT solution (final concentration: 0.01M); place in a cell culture incubator and incubate in the dark for 4 h; discard the culture medium, add 150 μL DMSO solution to each well, shake on a shaker at 100 r / min in the dark for 30 min, and The absorbance at OD570 nm was measured on an enzyme-linked immunosorbent assay.

[0049] In order to detect the effect of Metformin on the pr...

Embodiment 3

[0050] Example 3: Immunofluorescence staining detects the effect of Metformin on the content of BVDV dsRNA

[0051] Spread the cell slides on a 24-well cell culture plate, add 500 μL of 0.1% gelatin solution to coat for 30 minutes; discard the gelatin solution, and inoculate 1×10 5 MDBK cells were placed in a cell culture incubator; when the confluence of the cells reached 60%, the final concentrations of 0, 5, 10, and 20 mM Metformin were added to treat the MDBK cells for 8 hours; the culture medium containing the drug was discarded, washed once with PBS, and added 1000TCID 50 MDBK cells were treated with BVDV suspension for 2 hours and then replaced with 5% FBS cell culture medium; after 24 hours of infection, the culture medium was discarded, and gently washed twice with sterile PBS, 3min each time; 4% paraformaldehyde was added to fix at room temperature 15min; discard the fixative, wash with PBS 3 times, 3min / time; add blocking solution (0.5% TritonX-100, 1%BSA, 1%Donke...

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Abstract

The invention provides application of Metformin in preparation of a medicine for preventing and treating bovine enterovirus infection. The compound Metformin provided by the invention can effectively inhibit the proliferation of bovine enterovirus, experiments prove that the accumulation of mRNA and dsRNA of BVDV can be obviously inhibited by using Metformin to pretreat MDBK cells, the formation of virus particles can be obviously inhibited, the CPE phenomenon is obviously weakened, the final 10mM use effect of Metformin on the MDBK cells is better, BVDV challenge is performed after the Metformin is used for intragastric administration of a BALB / c mouse, the Metformin can obviously reduce the content of mRNA of the BVDV in visceral tissues of the mouse, through observation of pathological sections, the Metformin can effectively relieve tissue lesions caused by BVDV infection, which indicates that the Metformin inhibits replication of BVDV in cells and in mice, and the Metformin has high efficiency in the aspect of virus resistance. The Metformin has the prospect of being developed into the medicine for preventing and / or treating bovine enterovirus infection, a new medicine application is developed for the Metformin, an experimental foundation is laid for developing the efficient and specific anti-bovine enterovirus medicine, and a new view is provided for developing the efficient and specific anti-bovine enterovirus medicine.

Description

technical field [0001] The invention belongs to the technical field of the application of Metformin, specifically, the invention relates to the technical field of the application of Metformin in inhibiting bovine viral diarrhea virus infection. Background technique [0002] Bovine viral diarrhea / mucosal disease (BVD / MD) is a contagious infectious disease of cattle, sheep, pigs, deer and other animals caused by bovine viral diarrhea virus (BVDV). It can cause clinical symptoms such as high fever, diarrhea, chronic mucosal disease, and mucosal congestion. It can also cause a decrease in milk production in dairy cows, abortion in female animals, stillbirths or deformed fetuses, and reproductive disorders. In particular, non-cytopathogenic effect (NCP) BVDV infection in early pregnancy cows causes persistent infection (Persistent infection, PI) in calves, which carry the virus for life and detoxify to the outside world through nasal mucus, saliva, urine, and milk. , become a po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/155A61K45/06A61P1/12A61P31/14
CPCA61K31/155A61K45/06A61P1/12A61P31/14
Inventor 付强史慧君贺渊秀李泽宇陈俊贞杨莉冉多良
Owner XINJIANG AGRI UNIV
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