紫外线A诱导的人真皮成纤维细胞光老化及小鼠皮肤光老化模型的构建与评价

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[Abstract]ObjectiveTo establish photoaging modelsofhuman dermal fibroblasts（HDFs）and mouseskin induced byultravioletA（UVA），soas toprovidetheoptimizedmodelsforexploring mechanismsofskinphotoagingndpreventingandtreatingskin photoaging.MethodsEnzyme digestion was used to isolate HDFsand surface markers were examined.UVAatadose of 5 J/cm2 was utilizedtoirradiateHDFsfor15minuteseachdayforthreeconsecutivedays，andtheninvitroHDFphotoagingmodelswereestablished.Twelvefemale Kunming mice weredividedintoacontrol groupandamodel group（UVAiradiationgroup）withsix micein each group. A dose of 20J/cm2 of UVA was applied for 1 hour daily，5 times a week，which was continued for eight weeks to establish amouseskinphotoagingmodel.Furthermore，themodeleficacieswereaessedbybservingthecelllarmorphology，detectingthe cell proliferation ability and the activityof senescence-related β -galactosidase，observing the gross appearance of the dorsal skin of mice，performing hematoxylin-eosin staining and Masson staining of the skin tisues，detecting the expression of Pl6INK4a and P21 mRNA viaRT-qPCR，andexaminingtheexpressonoftypeIcollagen（Collagen-I）andmatrix metallprotenase1（MMP）byWesternblotandimmunohistochemicalstaining.ResultsHDFswerelongfusiformshaped，whichexpressedvimentinbutdidnotepress E-cadherin.AfterUVAiradiation，thesizeofHDFsbecamelargerwithspreading morphology，longwithaenuatedproliferatioability.And the positiverateof senescent cellsand the expressionlevels of Pl6INK4a and P21 mRNA were up-regulated. Meanwhile， Collagen-IexpressonasindhilexpreionwaslevatdCompardwiheontrolroup，thedoalsinficewas roughwithpigmentationafterUVAiradiation.UVAexposuregaverisetotickeningoftheepidermisandthiningofthederiswith decreased and disorganized collagen. Moreover，the expression levels of Pl6INK4a and P21 mRNA in UVA irrdiation group were dramaticallyhigherthantoseintecontrolgroup.AndteexpressionofCllge-Idminshed，whileeexpressoofMPnhanced.ConclusionThe UVA-induced HDFphotoaging modeland mouse skinphotoaging modelaresucessfullestablished，which provide an experimental basis for exploring the mechanism of skin photoaging and developing therapeutic strategies.

Keywords] Skinphotoaging；UltravioletA；Fibroblasts；Animal model；Evaluatiol

皮肤作为人体最大且位于最外层的器官，由表皮、真皮和皮下脂肪构成，在保护、排泄和感觉知觉方面发挥着关键作用[1]。（剩余12110字）