Fungal Keratitis (FK) is the main corneal infection that is resistant to antifugi drugs. Exploring new agent will be important for the drug resistance and new infectious diseases. All-Trans-Retinoic Acid (ATRA), a bioactive derivative of vitamin A, is immunoregulatary and anti-inflammation agent by decreasing the expression of the pro-inflammatory cytokines and increasing the expression of antiinflammatory factors. The study was to investigate the potential role of ATRA to be a supplementary method for the therapy of FK. The aim of this study was to investigate the role of ATRA Nanostructured Lipid Carriers (NLC) in the zymosan induced cytokines (IL-17, ICAM-1, MIP-2, MCP-1 and Ip-10) by Corneal Fibroblasts (CFs) and to investigate the characters of ATRA-NCL. ATRA-NLC was prepared by the method of Emulsification process from scratch. ATRA-NCL concentration was calculated by High Performance Liquid Chromatography (HPLC) method. ATRA-NCL physical stability was observed. ATRA-NCL Cytotoxicity assay was performed by the detection of Lactate Dehydrogenase (LDH). The role of ATRA-NCL on the release of IL-17, ICAM-1, MIP-2, MCP-1, IP-10 induced by zymosan was examined by ELISA. The relative NF-KB-p65, P-ERK1, 2, P-IκBα cell signal pathway was assayed by immune blot analysis. The mean diameter of ATRA-NLC was 200 nm. ATRA-NCL physical stability study showed well at 4°C without precipitation and crystallization. The NLC particles were dispersed well under the scanning electron microscope (S-4800). Scanning electron microscope image of the ATRA particles were shown. ATRA-NCL showed no cytotoxic effect on KCs. ATRA-NCL inhibited zymosan-induced IL-17, ICAM-1, MIP-2, MCP-1, Ip-10 release by CKs. ATRA-NCL inhibited NF-KBp65, P-ERK, P-IκBα signalling pathways induced by zymosan in KCs. ATRA-NLC can inhibit the releasing of cytokines (IL-17, ICAM-1, MIP-2, MCP-1, Ip-10) induced by zymosan in CFs. It was supplementary selection for the therapy of fungal keratitis.