红细胞膜仿生纳米颗粒通过减轻炎症反应在真菌性角膜炎中发挥保护作用

Red Blood Cell Membrane-Camouflaged Biomimetic Nanoparticle Plays a Protective Role in Fungal Keratitis by Reducing the Inflammatory Response

摘要: 目的：探究红细胞膜仿生纳米颗粒在真菌性角膜炎中的保护作用及机制。方法：首先通过角膜荧光素钠染色实验探究红细胞膜仿生纳米颗粒对小鼠角膜的毒性。在体内，用烟曲霉菌孢子感染C57BL/6小鼠角膜，构建烟曲霉菌性角膜炎小鼠模型，用PBS或者红细胞膜仿生纳米颗粒处理，用裂隙灯显微镜拍摄感染后1、3、5天角膜照片并进行临床评分，用RT-PCR检测感染后第3天角膜TNF-α、CCL-2的mRNA表达。在体外，用烟曲霉菌菌丝刺激HCECs后，用红细胞膜仿生纳米颗粒处理，通过RT-PCR检测HCECs中TNF-α、CCL-2的mRNA表达。结果：红细胞膜仿生纳米颗粒对小鼠角膜安全无毒，点眼后角膜未出现荧光素钠染色。较PBS对照组相比，红细胞膜仿生纳米颗粒组小鼠角膜溃疡程度显著减轻，炎症反应水平下降。红细胞膜仿生纳米颗粒降低TNF-α、CCL-2的mRNA表达。结论：红细胞膜仿生纳米颗粒通过减轻炎症反应在烟曲霉菌性角膜炎中发挥保护作用。

Abstract: Objective: To explore the protective effect and mechanism of red blood cell membrane-camouflaged biomimetic nanoparticle on fungal keratitis. Methods: Firstly, the toxicity of red blood cell membrane-camouflaged biomimetic nanoparticle to the cornea of mice was investigated by and corneal fluorescein sodium staining experiment. In vivo, a mouse model of Aspergillus fumigatus keratitis was constructed by infecting the corneas of C57BL/6 mice with Aspergillus fumigatus spores and treated with PBS or red blood cell membrane-camouflaged biomimetic nanoparticle. The photos of the corneas were taken with slit lamp microscope at 1, 3 and 5 days after infection and clinical scores were performed. Meanwhile, PT-PCR was used to detect the mRNA expression of TNF-α and CCL-2 in cornea on day 3 after infection. Then, in vitro, the HCECs were infected with Aspergillus fumigatus and treated with red blood cell membrane-camouflaged biomimetic nanoparticle. And RT-PCR was used to detect the mRNA expression of TNF-α and CCL-2 in HCECs. Results: The red blood cell membrane-camouflaged biomimetic nanoparticle was safe and non-toxic to mouse corneas, and there was no sodium fluorescein staining was observed in corneas after implantation. Compared with PBS control group, the red blood cell membrane-camouflaged biomimetic nanoparticle group significantly reduced the degree of corneal ulcer and the level of inflammation. Meanwhile, the red blood cell membrane-camouflaged biomimetic nanoparticle could decrease the mRNA expression of TNF-α and CCL-2. Conclusion: The red blood cell membrane-camouflaged biomimetic nanoparticle plays a protective role in Aspergillus fumigatus keratitis by reducing inflammatory response.