This study was aimed to investigate the anti-inflammatory effects of the chitosan/gelatin membrane cross-linked with genipin (GP). Fourier transform infrared spectrophotometry (FT-IR) was used to characterize the cross-linking reaction. The XTT assay was applied to evaluate the cytotoxicity of the chitosan/gelatin membrane with RAW264.7 cells. Production of nitric oxide (NO) was measured by the Griess colorimetric method. The gene expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and IL-6 were detected by quantitative real-time reverse-transcription polymerase chain reaction (real-time RT-PCR). These results suggested that the anti-inflammatory effects of genipin cross-linked chitosan/gelatin membrane might be the results from modulating the NO/iNOS pathway and inhibiting the mRNA expression of COX-2, IL-6 in activated macrophages.