The Regulatory Effect of miR-146a Overexpression on Corneal Inflammatory Response in a Mouse Model of Dry Eye Disease and Its Relationship with the IRAK1/TRAF6/NF-κB Signaling Pathway

ZHONG Jun-mu; LIN Xiao-yan; LAI Mei-hua; LIN Cui-rong

Abstract：Objective: To investigate the regulatory effect of miR-146a overexpression on corneal inflammatory response in a mouse model of dry eye, and to analyze its relationship with the IRAK1/TRAF6/NF-κB signaling pathway. Methods: A total of 50 SPF-grade BALB/c mice were randomly divided into five groups, with 10 mice in each group. Except for the control group, the other four groups were treated with 0.2% benzalkonium chloride (BAC) solution in both eyes to construct a dry eye model. After successful modeling, the control group and model group received NC agomir; the miR antagonist group received miR-146a antagomir; the miR agonist group received miR-146a agomir; and the pathway agonist group received miR-146a agomir + NF-κB activator 2. After four weeks of treatment, the expression levels of miR-146a, inflammatory factors, and IRAK1/TRAF6/NF-κB signaling pathway proteins were observed and compared among the five groups. Results: After four weeks of treatment, there was a statistically significant difference in the relative expression of miR-146a in the five groups (F=61.058, P<0.001), which was significantly higher in the miR agonist group than in the other four groups. After 4 weeks of treatment, there were statistically significant differences in the expression levels of IL-1β, IL-6, IL-8 and TNF-α in the five groups (F=84.757, 103.658, 55.477, 46.762; P<0.001). After four weeks of treatment, there were statistically significant differences in the protein expression levels of IRAK1, TRAF6, NF-κB and IκB α in the five groups (F=62.975, 77.173, 67.108, 29.381; P<0.001). Except for the control group, the expression levels of IRAK1, TRAF6 and NF-κB proteins in the miR agonist group were significantly lower than those in the other three groups, and the expression levels of IκBα protein were significantly higher than those in the other three groups. Conclusion: Overexpression of miR-146a can negatively regulate corneal inflammatory response in dry eye mice through the IRAK1/TRAF6/NF-κB signaling pathway, which can provide new insights for the clinical treatment of dry eye disease.