单纯疱疹病毒性角膜炎小鼠模型的建立及鉴定

Establishing and identifying of herpes simplex keratitis mice model

摘要:

目的：探讨建立不同感染时期HSK小鼠动物模型，为HSK的深入研究建立基础。

方法：Balb/c小鼠125只麻醉后在显微镜下用刀片背面尖端于角膜“#”字划痕，其中100只小鼠接种 HSV-Ⅰ病毒，另25只小鼠不接种病毒作为正常对照组。术后每天用10g/L荧光素钠染色后裂隙灯显微镜下观察角膜病变发生情况，并取角膜表面泪液进行HEK293T细胞检测以确定裂隙灯显微镜下有无病毒复制。对潜伏感染期小鼠模型采用紫外线B光照射以诱导HSK复发。

结果：接种HSV-Ⅰ病毒的小鼠模型眼于接种后3d内全部出现急性上皮性角膜炎表现。经阿昔洛韦滴眼液治疗1wk后角膜炎症消失，但角膜和三叉神经节中PCR检测病毒仍为阳性。潜伏感染期小鼠模型经紫外线B光照射后也都在1wk内复发，并表现为以基质型角膜炎为主要临床表现的角膜病变。

结论：采用角膜划痕法对Balb/c小鼠接种HSV-Ⅰ病毒和紫外线B光照射可以成功地制作出原发感染期、潜伏感染期和复发感染期等不同感染时期的HSK模型，而且操作相对简单、方便易行。

Abstract:

AIM: To establish a basis of in-depth study for herpes simplex keratitis through establishing different infection period herpes simplex keratitis mice model.

METHODS: Totally 125 Balb/c mice were scratched the word of "#" in cornea using the back of a razor blade cutting edge after anesthesia under the microscope. Among the total of 100 mice were inoculated with herpes simples virus type-Ⅰ. The rest of 25 mice were not inoculated with HSV-Ⅰas normal control group. Using 10g/L sodium fluorescein stained and slit lamp microscope observed the occurrence of corneal disease after operation everyday. Corneal surface tears were taken to carry on the detecting of human embryonic kidney epithelial cells(HEK293T)to define whether the viral replication. The latently infected mice model were exposed to ultraviolet-B light(UV -B)to induce the recurrence of HSK.

RESULTS: The eyes of 100 mice model which inoculated with HSV-Ⅰwere all appeared acute epithelial keratitis within 3 days after inoculation. The corneal inflammation disappeared after acyclovir eye drops treating for 1 week, but the virus detection of polymerase chain reaction(PCR)in cornea and trigeminal ganglion remained positive. The latently infected mice model were exposed to UV-B to induce the recurrence of HSK within 1 week, recurrent HSK were in the form of stromal HSV keratitis principally.

CONCLUSION: Balb/c mice were inoculated with HSV-I virus by corneal scarification, through UV-B rays establishing infected, latent, recurrent herpes simplex keratitis mice model, and the operation is relatively simple, convenient and easy.