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HaCaT keratinocytes to copy UVA-induced apoptosis model from of a acidic sphingomyelinase ( acidsphingomyelinase ASMase ) , c - Jun amino-terminal kinase ( c -Jun NH 2 -terminal kinase, JNK ) and cyclooxygenase-2 ( of cyclooxygenase - 2 , COX -2 ) point of view , research PCF (Polypeptide fromChlamys farreri, PCF) inhibit the molecular mechanisms of UVA - induced HaCaT apoptosis . Method to copy 8J · cm -2 < / sup > UVA irradiated HaCaT cells apoptosis model , according to the experimental design is divided into : control group , the UVA model group , positive control group (5.68 mmol · L -1 vitamin C, cell apoptosis was detected when the application of the group ) , the drug group (5.68 mmol · L -1 < / sup> PCF , 2.84 mmol · L -1 IPCF, 1.42 mmol · L -1 PCF), inhibitor group ( at 25μmol · L -1 Desipramine , 20μmol · L -1 < / sup> SP600125, 1μmol · L -1 celecoxib). Agarose gel electrophoresis analysis of the PCF, ASMase inhibitors ( Desipramine ) , JNK inhibitor (SP600125) and COX-2 specific inhibitor (celecoxib) on apoptosis ; with Hochest 33258 fluorescent staining UVA caused apoptosis morphological changes and the PCF apoptosis ; ASMase expression by RT-PCR and immunofluorescence staining ; Western blot ASMase, JNK, phosphorylated JNK protein expression ; Perfect Real Time PCR detection of c-jun mRNA expression ; RT-PCR to detect the expression of COX-2 combined with Western blotting . 1.42 ~~ 5.68 mmol · L -1 < / sup > dose range of PCF can significantly inhibit UVA - induced apoptosis of HaCaT cells ( P <0.05 ) of UVA - induced nuclear pyknosis apoptotic morphology change significantly inhibited ; pre join Desipramine , SP600125, and celecoxib can significantly inhibit the formation of UVA-induced HaCaT cell DNA Ladder ; PCF within the dose range of 1.42 ~~ 5.68 mmol · L (-1 ) dose-dependent inhibition of UVA - induced ASMase c -jun and COX-2 expression and JNK activation (P <0.05, P <0.01). Conclusion PCF inhibit 8J · cm -2 UVA -induced apoptosis of HaCaT cells , its mechanism of action to inhibit the activation of the pathway UVA -induced ASMase---JNK/c-jun---COX-2 of related.
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