Department of Ophthalmology, Far Eastern Memorial Hospital, No. 21, Sec. 2, Nanya South Road, Banqiao, New Taipei City 22056, Taiwan.
Department of Ophthalmology, National Taiwan University Hospital, No. 7, Chung-Shan South Road, Taipei 10002, Taiwan.
Mediators Inflamm. 2017;2017:9601349. doi: 10.1155/2017/9601349. Epub 2017 Dec 24.
This study aimed to determine the effect and roles of microRNA (miRNA, miR) treatment in experimental autoimmune anterior uveitis (EAAU).
Uveitis was induced in Lewis rats by simultaneous injections of bovine melanin-associated antigen into the hind footpad and the intraperitoneal cavity. The animals were injected intravitreally with low-dose (0.5 g) or high-dose (1.5 g) miR-146a. The clinical scores, leukocyte count in the aqueous humor, and histology were assessed. Cytokine changes were evaluated by relative mRNA expression and enzyme-linked immunosorbent assay (ELISA). The expression of nuclear factor kappa B (NF-B) was assessed by immunofluorescence and Western blotting. Evaluation of the DNA-binding activity of NF-B was performed by electrophoretic mobility shift assay (EMSA).
Treatment with miR-146a significantly attenuated clinical scores and leukocyte infiltration in a dose-dependent manner, a result that was compatible with histological findings. Following miR-146a injections, downregulation of interleukin- (IL-) 1, IL-6, and IL-12 and interferon- (IFN-) and upregulation of IL-10 and IL-17 were noted. The decreased NF-B expression on immunofluorescence and Western blotting and reduced DNA-binding activity on EMSA were demonstrated following miR-146a treatment.
miR-146a effectively reduced intraocular inflammation in EAAU through the inhibition of NF-B. miR-146a might be a new treatment choice for uveitis.
本研究旨在确定 microRNA（miRNA，miR）治疗在实验性自身免疫性前葡萄膜炎（EAAU）中的作用和机制。
通过将牛黑色素相关抗原同时注入后脚掌和腹腔，在 Lewis 大鼠中诱导葡萄膜炎。动物通过玻璃体内注射低剂量（0.5μg）或高剂量（1.5μg）miR-146a。评估临床评分、房水中白细胞计数和组织学变化。通过相对 mRNA 表达和酶联免疫吸附试验（ELISA）评估细胞因子变化。通过免疫荧光和 Western blot 评估核因子 kappa B（NF-κB）的表达。通过电泳迁移率变动分析（EMSA）评估 NF-κB 的 DNA 结合活性。
miR-146a 治疗以剂量依赖性方式显著减轻临床评分和白细胞浸润，这与组织学发现一致。miR-146a 注射后，白细胞介素（IL）-1、IL-6 和 IL-12 以及干扰素（IFN）-的下调和 IL-10 和 IL-17 的上调被观察到。miR-146a 治疗后，NF-κB 的免疫荧光和 Western blot 表达减少以及 EMSA 上的 DNA 结合活性降低。
miR-146a 通过抑制 NF-κB 有效减轻 EAAU 中的眼内炎症。miR-146a 可能成为葡萄膜炎的一种新的治疗选择。
