摘要
目的从p38促细胞分裂剂激活性蛋白激酶(p38MAPK)通路和半胱天冬酶-3(caspase-3)的角度,研究扇贝多肽(poly-peptidefromChlamysfarreri,PCF)抑制紫外线A波(UVA)引起的HaCaT细胞凋亡的分子机制。方法实验分为6组:对照组、UVA模型组、UVA+5.68mmol.L-1维生素C阳性对照组、UVA+5.69mmol.L-1PCF组、UVA+2.84mmol.L-1PCF组、UVA+1.42mmol.L-1PCF组。以正交实验设计确立UVA诱导HaCaT细胞凋亡模型;琼脂糖凝胶电泳分析PCF、p38MAPK抑制剂(SB203580)及caspase-3特异性抑制剂(Ac-DEVD-CHO)对细胞凋亡的影响;蛋白质印迹法检测p38MAPK及磷酸化p38MAPK表达;流式细胞术检测caspase-3的活性。结果PCF能明显抑制UVA引起的HaCaT细胞凋亡;SB203580和Ac-DEVD-CHO对UVA诱导的HaCaT细胞凋亡有抑制作用;1.425.69mmol.L-1内的PCF可剂量依赖性抑制UVA引起的p38MAPK磷酸化及caspase-3的活化。结论PCF可抑制UVA诱导的HaCaT细胞凋亡,其作用机制与抑制p38MAPK通路和caspase-3活性有关。
OBJECTIVE To investigate the inhibiton of polypeptide from Chlamys farreri （PCF） on UVA-induced HaCaT cells apoptosis through p38 mitogen activated protein kinase （MAPK） pathway and caspase-3. METHODS Experiments were divided into six groups： control group, UVA model group, UVA ＋ 5.68 mmol·L^-1 vitamine C positive control group, UVA ＋ 5.69 mmol·L^-1 PCF group, UVA ＋ 2.84 mmol·L^-1 PCF group, UVA ＋ 1.42 mmol·L^-1 PCF group. UVA-induced apoptotic model of HaCaT cells was established by orthogonal design. Using agarose gel electrophoresis, the effects of PCF, p38 MAPK inhibitor SB203580 and caspase-3 inhibitor Ac-DEVD-CHO on UVA-induced apoptosis were investigated. Expression levels of p38 MAPK and phosphorylated p38 MAPK were determined by Western blot analysis. Caspase-3 activity was assayed by flow cytometry. RESULTS PCF significantly protected UVA-induced apoptosis. SB203580 and Ac-DEVD-CHO had inhibitory effects on UVA-induced apoptosis of HaCaT cells. PCF inhibited UVA-induced phosphorylation of p38 MAPK and activation of caspase-3 on a dose-dependented manner. CONCLUSION PCF can protect HaCaT cells from UVA-induced apoptosis. Its inhibitory effect on apoptosis may attribute to inhibition of activation of p38 MAPK and caspase-3.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2007年第2期116-120,共5页
Chinese Pharmaceutical Journal
基金
国家自然科学基金(30471458)
山东自然科学基金(Y2003c02)
