Expressions of IL-6, TNF-α and NF-κB in the skin of Chinese brown frog (Rana dybowskii).

The cytokine interleukin-6 (IL-6) mediates a wide range of inflammatory and immune responses. Tumor Necrosis Factor α (TNF-α) has a myriad of pro-inflammatory effects on the skin. Nuclear factor κB (NF-κB) is a transcriptional factor that regulates a battery of genes that are critical to immune system. In this study, we investigated the localizations and expression levels of IL-6, TNF-α and NF-κB in the skin of Rana dybowskii during the breeding period and pre-hibernation. Histologically, the skin of Rana dybowskii consisted of epidermis and dermis. Four kinds of cells were identified in the epidermis, while the dermis was composed of homogenous gel, mucous glands and granular glands. IL-6, TNF-α and NF-κB were immunolocalized in the epithelial and glandular cells in both periods. Western blotting showed that IL-6, TNF-α and NF-κB were significantly higher in the pre-hibernation compared to the breeding period. Real- Time PCR revealed that the relative mRNA levels of IL-6 and NB-κB in the pre-hibernation increased significantly compared with the breeding period, while the TNF-α mRNA expression levels were not significantly different between these two periods. These results suggested that IL-6, TNF-α and NF-κB might collectively be involved in the skin immune system of Rana dybowskii during the breeding period and pre-hibernation.