扇贝多肽经由aSMase-JNK通路抑制UVA诱导HaCaT细胞凋亡  

Polypeptide from Chlamys Farreri Inhibits UVA-induced Apoptosis of HaCaT Cells Involving of aSMase-JNK Pathway

作　　者：邢雁霞[1] 苏爱[2] 杜卫[2] 王春波[3] 

机构地区：[1]青岛大学医学院生物化学与分子生物学教研室 [2]青岛大学医学院细胞与分子生物学实验室,青岛266071 [3]青岛大学医学院药理教研室,青岛266071

出　　处：《高等学校化学学报》2008年第4期757-761,共5页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:30471458);国家“八六三”计划(批准号:2003AA625070)资助

摘　　要：建立紫外线A（UVA）辐射损伤HaCaT细胞的病理模型，从酸性鞘磷脂酶-JNK信号通路的角度研究扇贝多肽（Polypeptide from Chlamys farreri，PCF）抑制UVA诱导HaCaT细胞凋亡的分子机制．采用Hoechst 33258染色结合琼脂糖凝胶电泳分析细胞凋亡；用RT—PCR法和细胞免疫荧光染色检测胞内酸性鞘磷脂酶（acid sphingomyelinase，aSMase）的表达；蛋白印迹法检测细胞内JNK及磷酸化JNK的蛋白水平．结果表明，PCF可明显地抑制UVA诱导的HaCaT细胞凋亡；aSMase抑制剂Desipramine和JNK抑制剂SP600125均可阻断UVA引起的细胞凋亡；PCF的浓度在1．42～5．68mmol／L范围内可依赖性地抑制UVA辐射后细胞内aSMase的表达量以及JNK蛋白的磷酸化；预先加入Desipramine则抑制UVA引起的JNK蛋白的磷酸化．表明PCF通过阻断aSMase—JNK通路来抑制UVA诱导HaCaT细胞凋亡．A pathological model of UVA-induced HaCaT cells was established to investigate whether polypeptide from Chlamysfarreri（PCF） protects HaCaT cells from apoptosis induced by UVA through acid sphingomy- elinase （aSMase） -JNK pathway and to explore its related molecular mechanism. Apoptosis of cells were determined by Hoechst 33258 staining and agarose gel electrophoresis. The expression of aSMase in HaCaT cells was detected via RT-PCR and immunofluorescence. Using Western Blot analysis, expression levels of JNK and phosphorylated JNK were assayed. The results showed that PCF can significantly protect against UVA-induced apoptosis of HaCaT cells. PCF can also inhibit expression of aSMase and phosphorylated of JNK in HaCaT cells radiated by UVA in a doseependent manner. ASMase inhibitor Desipramine and JNK inhibitor SP600125 had inhibitory effects on UVA-induced apoptosis, the former blocked phosphorylated of JNK. So it is concluded that PCF can protect HaCaT cells from apoptosis induced by UVA and its protective effects may attribute to blocking aSMase-JNK pathway.

关 键 词：扇贝多肽 酸性鞘磷脂酶 JNK 紫外线A 凋亡 HACAT细胞 

分 类 号：Q514[生物学—生物化学] Q257