TGF-β1-induced Artificial Skin To UV Light Damage Inflammatory Cytokine IL-1α, IL-6, TNF-α Impact Study

Objective:Making artificial skin on exposure to simulated UV light damage model, to intervene in TGF-β1, TGF-β1 on the study of artificial UV-induced skin light injury.Method:1.To cultivate artificial skin, HE staining:Cultured fibroblasts and keratinocytes to collagen matrix, the construction of artificial skin;2.The establishment of light damage model:MTT assay of cell activity of UV damage, selection of appropriate UV dose;.3.TGF-β1-induced artificial skin to UV light damage model of intervention:the artificial skin were divided into negative control group, UV irradiation, TGF-β1 i intervention group, UV TGF-β1 intervention group;4.HE staining:to observe the morphology of artificial skin changes in each group;5.ELISA (Enzyme-linked immunosorbent assay, ELISA):to detect the intervention group after 24h cytokine IL-1α, IL-6, TNF-αsecretion of the situation;Results:1.HE staining results showed:Artificial skin from the epidermis and dermis layers, reticular dermis arranged, dyed pink collagen, during interspersed evenly distributed fibroblasts. Epidermis from the layered composition of keratinocytes, basal keratinocytes were columnar, tight connection between cells,3-4 layers visible on its round, oval closely connected keratinocytes2.Construction of light damage model:comparison with MTT method was found 30J/cm~2 inhibition rate irradiation there were significant differences (p<0.01);3.HE staining showed:Compared with the negative control group, UV irradiation epidermal layer disappeared, the cell gap widened, with nucleus fragmentation, showing apoptosis. Dermal layer of loose collagen matrix structure changes, fibroblasts reduced. TGF-β1 in the intervention group have no significantly changes. UV TGF-β1 intervention group epidermal layer disappeared, the cell gap widened, nucleus fragmentation, cell apoptosis, dermal collagen matrix structure, loose change, fibroblasts reduced.4.ELISA assay showed:Compared with negative group,UV irradiation disease factor IL-1α, IL-6, TNF-αalso significantly increased, there was significant difference (p<0.01). UV TGF-β1 intervention group IL-1α, IL-6, TNF-αwas significantly increased, there was significant difference (p<0.01). However, UV TGF-β1 intervention group compared with the UV radiation is the IL-1α, IL-6, TNF-αdecreased (p<0.01).Conclusions:1. Successfully constructed artificial skin;2.With 30J/cm~2 (UVA and UVB)build a successful model of light damage;3. For the first time in the light damage model of artificial skin based on, showed that TGF-β1 inhibit inflammatory factors IL-1α, IL-6, TNF-αproduction of the artificial skin to UV light damage model, may be the protective effect of light damage model.The mechanism of light damage and the development of new drug targets for selection of some significance...