Zhang Li, Jiang Nan, Zhao Gui-Qiu, Peng Xu-Dong, Zhu Guo-Qiang, Jiang Wei, Ma Jing-Jing
Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.
Int J Ophthalmol. 2020 Feb 18;13(2):199-205. doi: 10.18240/ijo.2020.02.01. eCollection 2020.
To observe the expression and role of aryl hydrocarbon receptor (AhR) in the immune response of mouse cornea infected with .
Murine models of keratitis were established by scraping the central epithelium of mouse cornea, daubing on the cornea and covering with a contact lens. The mice were randomly divided into the control group and the -infected (A.F.) group for 1, 3 and 5d respectively, which corneas were daily monitored by a slit lamp microscope and the clinical scores were also recorded timely after infection. In this study, immunofluorescence staining was used to detect the expression and localization of AhR in mouse corneas, and the mRNA and protein of AhR were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. In addition, mouse peritoneal macrophages were stimulated by with or without the pretreatment of AhR antagonist CH223191 and AhR agonist FICZ, and the tumor necrosis factor alpha (TNF-α), inducible nitric oxide synthase (iNOS), interleukin-10 (IL-10) and Arg-1 mRNA were detected by RT-PCR.
According to the results of the slit light photography, it was clearly indicated that the corneal inflammation were the most severe and the clinical score became the highest as well on the 3 day after the infection of . Contrasted with the control group, the expression of AhR in the corneal epithelial cells infected with was significantly increased detected by immunofluorescence staining. AhR mainly expressed in the nucleus and cytoplasm of corneal epithelial cells. Consistent with the transcriptional level of AhR mRNA, the expression level of AhR protein reached the peak on the 3 day after infection which was detected by Western blot. Furthermore, RT-PCR showed that CH223191 up-regulated the expression of TNF-α and iNOS and down-regulated the expression of IL-10 and Arg-1 in peritoneal macrophages; inversely, FICZ reduced the expression of TNF-α and iNOS while elevated the expression of IL-10 and Arg-1.
AhR is involved in the pathogenesis of keratitis and induced immune protection in anti- immune response by inhibiting M1 and increasing M2 phenotype macrophage-related inflammatory factors.
观察芳烃受体（AhR）在感染 的小鼠角膜免疫反应中的表达及作用。
通过刮除小鼠角膜中央上皮、在角膜上涂抹 并覆盖隐形眼镜建立小鼠角膜炎模型。将小鼠随机分为对照组和 感染（A.F.）组，分别于感染后1、3和5天进行观察，每天用裂隙灯显微镜监测角膜情况，并在感染后及时记录临床评分。本研究采用免疫荧光染色检测AhR在小鼠角膜中的表达及定位，通过逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹法检测AhR的mRNA和蛋白。此外，用 刺激小鼠腹腔巨噬细胞，同时用AhR拮抗剂CH223191和AhR激动剂FICZ进行预处理，通过RT-PCR检测肿瘤坏死因子α（TNF-α）、诱导型一氧化氮合酶（iNOS）、白细胞介素-10（IL-10）和精氨酸酶-1（Arg-1）的mRNA。
根据裂隙灯摄影结果，明确显示感染 后第3天角膜炎症最严重，临床评分也最高。免疫荧光染色检测显示，与对照组相比，感染 的角膜上皮细胞中AhR的表达明显增加。AhR主要表达于角膜上皮细胞的细胞核和细胞质中。蛋白质印迹法检测结果与AhR mRNA转录水平一致，感染后第3天AhR蛋白表达水平达到峰值。此外，RT-PCR显示CH223191上调了腹腔巨噬细胞中TNF-α和iNOS的表达，下调了IL-10和Arg-1的表达；相反，FICZ降低了TNF-α和iNOS的表达，同时升高了IL-10和Arg-1的表达。
AhR参与 角膜炎的发病机制，并通过抑制M1型巨噬细胞和增加M2型巨噬细胞相关炎症因子在抗 免疫反应中诱导免疫保护。
