扇贝多肽在中波紫外线诱导的HaCaT细胞凋亡中对p38MAPK-HSP27通路的影响

Effects of polypeptide from Chlamys farreri on p38MAPK-HSP27 pathways in UVB-induced apoptosis of HaCaT cells

目的从p38MAPK-HSP27通路方面研究扇贝多肽（polypeptide from Chlamys farreri,PCF）抑制中波紫外线（UVB）诱导的HaCaT细胞凋亡的作用机制。方法复制UVB诱导的HaCaT细胞凋亡模型;实验设计为6组：对照组、UVB模型组、UVB＋5.69mmol.L-1维生素C阳性对照组、UVB＋5.69mmol.L-1PCF组、UVB＋2.84mmol.L-1PCF组、UVB＋1.42mmol.L-1PCF组。PCF预处理30min,荧光染色（Hoechst33258）结合琼脂糖凝胶电泳分析PCF在p38抑制剂（SB203580）存在和不存在两种条件下对UVB诱导的HaCaT细胞凋亡的影响;蛋白质印迹法检测p38MAPK、HSP27的表达及磷酸化水平的改变,同时分析HSP27在细胞内定位的改变。结果PCF可明显抑制UVB诱导的HaCaT细胞凋亡;PCF在1.42~5.69mmol.L-1范围内,可剂量依赖性抑制p38MAPK和HSP27的磷酸化水平,并抑制HSP27由胞质向胞核中的移位。结论PCF通过阻断p38MAPK-HSP27通路来抑制UVB诱导的HaCaT细胞凋亡,其作用机制与抑制p38MAPK的活化从而阻止HSP27的磷酸化及其细胞内移位有关。

Aim To investigate whether polypeptide from Chlamys farreri（PCF） protected HaCaT cells from UVB-induced apoptosis through p38 MAPK-HSP27pathway. Methods UVB-induced apoptotic model of HaCaT cells was copied by the preliminary study. Experiment design was divided into six groups：control group, UVB model group, UVB ＋ 5.69 mmol· L^-1 vitamin C positive control group, UVB ＋ 5.69 mmol· L^-1 PCF group, UVB ＋ 2. 84 mmol · L^-1 PCF group and UVB ＋ 1.42 mmol · L^-1 PCF group. The HaCaT cells were treated with PCF for 30 rain, using fluorescence staining（ Hoechst 33258 ）and agarose gelelectrophoresis,the effects of PCF on UVB-induced apoptosis of HaCaT cells were investigated in each group with or without p38MAPK inhibitor （SB203580）. The expression levels of p38 MAPK, phosphorylated p38 MAPK （ p-p38 ） and HSP27, phosphorylated HSP27 （ p- HSP27）, the expressions of HSP27 in cytosolic fraction and nuclear fraction were also detected by Western blot. Results PCF significantly protected against UVB-indueed apoptosis of HaCaT cells. PCF inhibited UVB-induced phosphorylation of p38 MAPK, HSP27 and translocation of HSP27 from the cytoplasm to nu- cleus. Conclusions PCF inhibits UVB-indueed apoptosis of HaCaT cells through p38 MAPK-HSP27 path- way. Its inhibitory effeet on apoptosis may attribute to dose-dependently inhibited phosphorylation of p38, HSP27 and transloeation of HSP27 from the cytoplasm to nucleus.