【摘要】 目的探讨扇贝多肽(PCF)对中波紫外线(UVB)辐射损伤HaCaT细胞的作用及对细胞外调节激酶-丝裂原活化蛋白激酶(ERKs/MAPKs)通路的影响。方法建立UVB(20 mJ/cm2)对体外培养的HaCaT细胞辐射损伤的病理模型,实验分为对照组、模型组、UVB+5.69 mmol/L PCF组、UVB+2.84 mmol/L PCF组、UVB+1.42 mmol/L PCF组和UVB+5.69 mmol/L维生素C组。琼脂糖凝胶电泳观察各组不加入或分别加入Caspase-3抑制剂(DEVD-FMK)、ERKs通路抑制剂(PD98059)后的细胞凋亡情况;Western blot检测ERKs、MEKs磷酸化与非磷酸化的活性。结果PCF能剂量依赖性地减少UVB所致的HaCaT细胞DNA片段的出现;预先应用ERKs通路抑制剂可使DNA片段增加;预先应用Caspase-3抑制剂使DNA片段减少;PCF还能提高磷酸化ERKs、MEKs的活性,但不影响总的ERKs、MEKs的含量。结论PCF能抑制UVB辐射诱导的细胞凋亡,对UVB辐射损伤的细胞具有保护作用。其作用通过调节ERKs/MAPKs的信号通路和Caspase级联反应实现。更多还原

【Abstract】 Objective To investigate whether PCF could inhibit apoptosis of HaCaT cells induced by ultraviolet B and to examine the effect of PCF on the ERKs/mitogen-activated protein kinases(ERKs/MAPKs) cascade and Caspase-3 in vitro.(Methods Cellular) experiments were designed into 6 groups(control,model,UVB+5.69 mmol /L PCF,UVB+2.84 mmol/L PCF,UVB+1.42 mmol/L PCF and UVB+5.69 mmol/L Vit C).Gel electrophoresis was used to observe the DNA ladder of each group with or without Caspase-3 inhibitor,ERKs pathways inhibitor;and Western blot was used to detect the activity of p-ERKs,P-MEKs,ERKs,and MEKs. Results PCF inhibited UVB-induced DNA ladder in HaCaT cells;ERKs pathways inhibitor increased the fragment of DNA by UVB;but the inhibition of Caspase-3 reduced the fragment of DNA;PCF enhanced the activity of P-ERKs and P-MEKs,but did not affect the activity of ERKs or MEKs. Conclusion PCF has protective effects against UVB-induced apoptotic cell death in HaCaT cells and PCF is likely to exert its cytoprotective effect in HaCaT cells through ERKs/MAPKs activation and caspase cascade.更多还原