Tree shrew herpes simplex keratitis (HSK) model building and evaluating method and application thereof

A technology for viral keratitis and herpes simplex, which is applied in the field of animal models of human diseases, can solve problems such as small corneas, large differences in immune responses, and difficult surgical operations, and achieve high success rate, simple operation, and good stability

Inactive Publication Date: 2016-05-04
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, rodents such as mice and rabbits are the main model animals used for the research of herpes virus keratitis, but because the corneas of mice are very small, it is difficult to perform surgical operations, and there are few specimens that can be obtained after infection, and they are not available. Facilitate the observation of the gross morphology of virus infection
Furthermore, the mouse model lacks clinical reactivation capacity for the virus
The immune response between the rabbit model and human infection is very different, and the cost of rabbit experiments is high

Method used

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  • Tree shrew herpes simplex keratitis (HSK) model building and evaluating method and application thereof
  • Tree shrew herpes simplex keratitis (HSK) model building and evaluating method and application thereof
  • Tree shrew herpes simplex keratitis (HSK) model building and evaluating method and application thereof

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Experimental program
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Embodiment 1

[0036] Embodiment 1 A kind of construction method of tree shrew herpes simplex virus keratitis model

[0037] The construction method of a kind of tree shrew herpes simplex virus keratitis model of the present invention is that HSV-1 virus is infected with HSV virus through tree shrew eye inoculation, establishes tree shrew herpes simplex virus keratitis model, comprises the following steps:

[0038] (1) Experimental animal preparation: prepare female adult healthy tree shrews aged 6 months to 3 years;

[0039] (2) Anesthesia and treatment of the tree shrew: fix the tree shrew in a special small cloth bag, weigh and record its weight, inject ketamine hydrochloride into the thick muscle of the inner thigh of the tree shrew, and the dosage of ketamine is determined according to the tree shrew The body weight is 50mg / kg, and after 5 minutes, the tree shrew is in a state of anesthesia;

[0040] (3) Virus infection: Scratch the eyeball surface of the tree shrew ten times horizonta...

Embodiment 2

[0041] Embodiment 2 A kind of construction method of tree shrew herpes simplex virus keratitis model

[0042] Other parts are identical with embodiment 1, difference is:

[0043] In the experimental method, the consumption of ketamine (ketamine) in the step (2) is 100 mg / kg by tree shrew body weight;

[0044] Step (3) in the experimental method: directly convert 1×10 6 The PFU dose of HSV-1 McKrae virus strain was dripped onto the two eye membranes of the tree shrew with a pipette gun to obtain the tree shrew herpes simplex virus keratitis model.

[0045] In addition, in the experiment, the inventor also set up a control group of tree shrew animals, that is, only scratching the eye mask of the animal, without adding the virus, and dropping an equal volume of PBS solution to the tree shrew eye mask (HSV-117 + control group); or without scratching the animal eye mask, directly drop an equal volume of PBS solution to the tree shrew eye mask (HSV-1McKrae control group).

[0046...

Embodiment 3

[0058] Example 3 Evaluation method and application of a tree shrew herpes simplex virus keratitis model

[0059] The present invention uses methods such as slit lamp examination, immunohistochemistry, live virus titer detection, viral genome load determination, and in situ hybridization to identify and evaluate the infection, latency and spontaneous reactivation of HSV-1 in tree shrew eyes. These experimental results All indicated that HSV-1 can effectively infect tree shrew eyes, and can better simulate the symptoms of human herpesvirus keratitis, and the structure of tree shrew cornea is highly similar to human cornea. The establishment of this keratitis model is conducive to further research on the occurrence, development and recurrence of herpes virus keratitis, as well as the mechanism of virus latency and reactivation, and has great value and potential in the application research of the development of antiviral drugs. Therefore, the tree shrew herpes viral keratitis mode...

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Abstract

The invention discloses a tree shrew herpes simplex keratitis (HSK) model building and evaluating method and application. Tree shrews are inoculated and infected with herpes simplex viruses (HSV-1) through eyes, a tree shrew HSK model is built, infection, virus incubation and reactivation of tree shrew eye tissue are researched and analyzed through silt-lamp examination, qPCR, qRT-PCR, immunohistochemistry, in-situ hybridization, in-vitro co-culture experiments, tissue live virus detection and viral genome quantification and other methods, and effectiveness and practicability of the tree shrew HSK model are identified and assessed. The tree shrew HSK model is built successively for the first time, infection of human being viral keratitis can be simulated better, and a better experimental animal model can be provided for a herpes simplex keratitis infection mechanism, antiviral drug evaluation and treatment research. The animal model is good in repeatability and stability; the building and evaluating method is easy to operate, the technology and equipment requirement is low, and the success rate is high.

Description

technical field [0001] The invention relates to a construction, evaluation method and application of a tree shrew herpes simplex virus keratitis model, belonging to the technical field of human disease animal models. technical background [0002] Herpes simplex virus (HSV) is an infectious virus that exists widely in nature and is susceptible to human infection. More than 80% of the global population has been infected with herpes simplex virus. Herpes simplex virus is divided into two genotypes, namely type I and type II herpes simplex virus Virus. Type 1 herpes simplex virus (HSV-1) infection site is the head and neck, which can cause a variety of human diseases, including herpes labialis, gingivostomatitis, keratitis, and even encephalitis. Type II affects the genitals and occasionally causes eye infections. Type Ⅱ infection in newborns, in addition to involving the eyes can also spread to the skin, blood, internal organs and central nervous system, and can be fatal. Hu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00
CPCA61K49/0008
Inventor 李丽红周巨民陈桂俊杨卫郎丰超杨丽萍王二林李欣李卓然
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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