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Jin Yuan, Lan Huang, Jing Zhong, State Key Opthalmic Laboratory; Tacrolimus (FK506) suppresses TREM-1 Expression in a murine model of Fungal Keratitis. Invest. Ophthalmol. Vis. Sci. 2014;55(13):6286.
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To investigate whether tacrolimus suppresses triggering receptor expressed on myeloid cells-1(TREM-1) expression in fungal keratitis and its effects.
RAW264.7 macrophages were devided into four groups: group i received zymosan(100ug/ml),group ii received zymosan (100ug/ml)+mTREM-1/Fc(20uM), group iii received zymosan(100ug/ml)+FK506(20uM), and group iv was the control and received no treatment. The expression of TREM-1,Interleukin-1β(IL-1β) and tumor necrosis factor alpha(TNFα) were assayed at 0h,4h,8h,12h,24h and 48h after zymosan treatment using real-time reverse transcriptase polymerase chain reaction (RT-PCR). The protein levels of sTREM-1,IL-1β,and TNFα were tested using ELSIA. An Aspergillus Fumigatus spore preparation (AFSP) was injected intrastromally to establish the C57BL/6J(B6) mouse model of fungal keratitis.Then, the mice were randomized into 2 groups: group A received four doses of FK506 eye drops each day, and group B received four doses of menstruum eye drops each day. The expression of TREM-1,IL-1β and TNFα was determined using real-time RT-PCR and ELISA at different time points. Myeloperoxidase(MPO) protein levels were confirmed by ELISA. Corneal damage was evaluated by clinical scores and histologic examination.
FK506 reduced the expression of TREM-1,IL-1β and TNF-α in RAW264.7 macrophages stimulated by zymosan. TREM-1 expression was significantly reduced in FK506-treated mouse corneas with fungal keratitis at 1 day and 3 days post-infection (P<0.05). There was no change detected at 5 days post-infection between the two groups. The expression levels of IL-1β and TNF-α presented the same trend as TREM-1 at 1 day post infection (P<0.05). The clinical score of group A at 1 day p.i.was 2.8 ±0.63, and the clinical score of group B was 2±0.66 (P<0.05).There was no statistically significant difference between the two groups at 3 and 5 days p.i. Hematoxylin-eosin(HE) staining and MPO ELISA data indicated that polymorphonuclear neutrophil(PMN) infiltration was diminished in group A at 1 day p.i.(P<0.001).The CFU data indicated a significant decrease in fungal viability at 1 and 3 days p.i. in group A (P<0.05).
FK506 suppressed the inflammatory response in a fungal keratitis mouse model by down-regulating TREM-1,IL-1β and TNF-α expression and reducing the infiltration of PMNs.
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