肉毒杆菌B诱导大鼠干眼模型中泪腺炎症因子表达研究

Study on the expression of inflammatory factors in the botulium toxin B-induced rats dry eye model

摘要:

目的：建立肉毒杆菌B诱导大鼠炎症干眼模型，通过观察泪腺炎症因子表达变化为干眼病实验研究提供实验依据。

方法：健康8周龄雌性SD大鼠，30只，随机分为2组，20只为实验组，10只为对照组。右侧泪腺注射肉毒杆菌B 0.1mL(20mU)，生理盐水0.1mL。分别于实验前1d及实验后第3d； 1，4，6wk，观察基础泪液分泌量、角膜荧光染色，并且酶联免疫吸附试验(ELISA)检测泪腺组织中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和巨噬细胞移动抑制因子(MIF)变化。

结果：术后实验组从第3d时出现泪液分泌减少，4wk达到高峰，6wk后恢复； 角膜荧光染色第3d时出现，持续6wk； 泪腺组织IL-1β从第3d开始出现增加，持续6wk； TNF-α表达水平未增加，MIF检测从第4wk开始表达增加，持续6wk。

结论：泪腺注射肉毒杆菌B可以成功诱导大鼠炎症干眼模型，炎症因子的表达变化具有一定的特征性，为干眼病实验提供较理想的动物模型。

Abstract:

AIM: To explore the changes of macrophage migration inhibitory factor(MIF), interleukin 1(IL-1β), tumor necrosis factor-alpha(TNF-α)levels in the botulium toxin B-induced murine dry eye model.

METHODS: The rats were randomly divided into two groups, 20 mice received injection of botulium toxin B(0.1mL), and 10 mice were injected physiological saline(0.1mL). Schirmer I test, corneal fluorescein staining and the levels of MIF, IL-6,TNF-α were performed before and 3, 7, 28, 42 days after injection. The levels of MIF, IL-6, TNF-α were detected by enzyme-linked immunosorbent assay(ELISA).

RESULTS: The tear production was significantly decreased at 4 points and the corneal fluorescein staining increased at 5 points in BTX-B-injected mice compared with control mice. In the BTX-B-injected mice, the level of IL-1β increased significantly at the 3 days and 1, 4, 6 week, and the level of MIF in lacrimal gland increased significantly since the 4^th week compared with control mice. The level of TNF-α has no difference between the two groups.

CONCLUSION: Injection botulium toxin B can successfully established the mice model of dry eye. This model has the characteristic changes of the expression of inflammatory factors, which is an ideal animal model for dry eye experiment.