蒙花苷通过调节NLRP3炎症小体改善实验性干眼模型的先天免疫炎性反应
Linarin ameliorates innate inflammatory response in an experimental dry eye model via modulation of the NLRP3 inflammasome
摘要
Abstract
Objective To investigate the effect and mechanism of linarin (LA) in an experimental dry eye model. Methods LA or vehicle was applied in two dry eye models: an in vitro hyperosmotic stress model and an in vivo desiccating stress (DS) murine model. The viability of human corneal epithelial cells (HCECs) was measured using a cell counting kit (CCK-8). Tear secretion was assessed using the phenol red cotton test. The tear break-up time (TBUT) was recorded using 0.1％ liquid fluorescein sodium. Corneal epithelial permeability was evaluated through Oregon green dextran (OGD) staining. Conjunctival goblet cells were counted using periodic acid-Schiff (PAS) staining. Terminal deoxynucleotidyl transfer dUTP nick-end labeling (TUNEL) staining was used to quantify apoptotic cells in both models. The expression of Ki-67 was measured in HCECs in the cell model while that of matrix metalloproteinase (MMP)-3 and -9 was measured in the murine model through immunofluorescence staining. Real-time quantitative PCR (RT-qPCR) was performed to assess the expression of MMP-3 and MMP-9 in the corneal epithelium and NLRP3, ASC, Caspase-1, interleukin (IL)-1β, IL-18, and tumor necrosis factor (TNF)-α in the conjunctiva. The protein expression levels of NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the conjunctiva were assessed via Western blot. Results In the in vitro model, treatment of HCECs with LA showed no toxicity, increased proliferation, and reduced apoptosis. In the murine model, compared to the control, LA significantly increased tear production and TBUT, improved OGD staining, and increased the number of goblet cells. Topical treatment of LA to mice provided decreased expression of MMP-3, MMP-9, TNF-α, and apoptotic corneal epithelium. Topical administration of LA also suppressed the NLRP3 inflammasome in the dry eye disease (DED) murine model by decreasing the expression of NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the conjunctiva.Conclusion Our findings support the safety and efficacy of LA in the treatment of DED. LA alleviated corneal epithelial damage and suppressed NLRP3 inflammasome-mediated immunity in the conjunctiva in a murine model of DED.关键词
蒙花苷/干眼病/NLRP3炎症小体/眼表/先天免疫炎性反应Key words
Linarin/Dry eye disease/NLRP3 inflammasome/Ocular surface/Innate inflammatory response引用本文复制引用
陈梅,彭清华,李洁,彭俊,黄雨,欧阳维杰,刘晓清,沈志斌,李常栋,王一..蒙花苷通过调节NLRP3炎症小体改善实验性干眼模型的先天免疫炎性反应[J].数字中医药（英文）,2021,4(1):42-53,12.基金项目
We acknowledge the assistance and valuable suggestions provided by Prof. LIU Zuguo and Mr. NAN Kaihui for this research. We thank for the funding support from the China Postdoctoral Science Foundation Grant (No. 2018M632973), Sichuan Science and Technology Program (No. 2018JY0388), the First-Class Open Fund for Integrated Chinese and Western Medicine (No. 2018ZXYJH05), Traditional Chinese Medicine First-Class Discipline Open Fund (No. 2018ZYX57), the Construction Project of the Hunan Engineering Technology Research Center for the Prevention and Treatment of Otorhinolaryngo-logic Diseases and Protection of Visual Function with Chinese Medicine (No. 2018YGC02 and No. 2018YGC04), the Research and Innovation Project of Graduate Students in Hunan Province (No. CX20190538). （No. 2018M632973）
