Expression Of Immune Cytokines In The Skin Of Ranu Dybowskii

Rana dybowskii is an important food and medicinal economic animal in China.Its skin contains a large number of biologically active substances.It has a special molecular structure and complex and diverse functions,and has great development potential and important application value.At present,Rana dybowskii skin products have been developed and utilized in the food industry.For example,its extracts are used as preservatives and health products for fruits and vegetables.Rana dybowskii skin plays an important role in defense against the immune defense against pathogenic microorganisms,and cytokines are important regulatory factors in the immune response.This study mainly studied the expression of immune factors in the skin of Rana dybowskii,and provided basis for elucidating the immune regulatory function of Rana dybowskii It is of great significance for the scientific,effective,and rational use of Rana dybowskiiIn this study,Rana dybowskii was selected as the object of study,and dorsal skin tissue was taken to examine the concentrations of IL-1?,IL-6 and TNF-a by enzyme linked immunosorbent assay and the expressions of IL-1?,IL-6 and TNF-a using immunohistochemistry,Western blotting and Real-Time PCR analysis.In order to explore the pathways by which these cytokines are generated,we evaluated the expression changes of the pathogen associated molecular pattern receptors(TLR4)and MyD88 signaling pathway factors(MyD88,IRAK4 and NF-?B)in dorsal skinThe results showed:(1)the skin of Rana dybowskii consists of epidermis and dermis.The epidermis is composed of stratum corneum,stratum granulosum,stratum spinosum and stratum germinativum.The dermis is distributed with stratum spongiosum and stratum compactum Pigment cells are located between the epidermis and the dermis.(2)Immunohistochemistry showed that IL-1?,IL-1R1,IL-6 and TNF-? were mainly expressed in epidermal cells and glandular cells of the Rana dybowskii skin during breeding period and pre-hibernation.Western blotting results showed that the expressions of IL-1? had no significant difference during these two period.Expressions of IL-1R1,IL-6 and TNF-a were significantly higher in the pre-hibernation.Ezyme linked immunosorbent assay detected the concentrations of IL-1?,IL-6 and TNF-a.Real-Time PCR analysis proved that the mRNA expressions of IL-1,IL-6,TNF-a and mRNA in the skin.(3)TLR4,MyD88,IRAK4 and NF-?B were expressed at the protein and molecular levels,and the expression of NF-?B in the skin was significantly increased in the pre-hibernation.In conclusion,the Rana dybowskii skin has the ability to secrete IL-1?,IL-6 and TNF-? and can synthesis IL-1?.The expressions of TLR4,MyD88,IRAK4 and NF-?B showed that the production of the three cytokines may be regulated by the TLRs and MyD88 signaling pathways,and ultimately participate in the inflammatory process and modulate the immune response.