摘要
目的观察佛波酯(PMA)慢性处理大鼠血管平滑肌细胞各亚型蛋白激酶C(PKCs)表达的影响。方法原代培养大鼠血管平滑肌细胞,①按照PMA处理浓度将细胞随机分为空白组、0.25‰DMSO组和PMA1、5、10μmol/L组,各组细胞均处理4 h;②按照PMA处理时间将细胞随机分为空白组、0.25‰DMSO组和PMA 1、4和24 h组,PMA组的药物浓度均为10μmol/L。采用Western blotting技术检测各亚型PKCs蛋白的表达。结果 PMA浓度<10μmol/L处理细胞时间<4 h不影响PKC-α的表达(P>0.05),10μmol/L PMA处理24 h能抑制PKC-α的表达(P<0.05);PMA浓度>5μmol/L处理细胞时间>4 h可明显抑制PKC-δ的表达(P<0.01),PMA浓度<5μmol/L处理细胞时间<4 h对PKC-ε的表达无显著影响(P>0.05),10μmol/L PMA处理细胞1 h即可明显抑制PKC-ε的表达(P<0.01);PMA浓度>5μmol/L处理细胞时间>4 h可以明显抑制PKC-θ的表达(P<0.01);10μmol/L的PMA处理细胞24 h对PKC-ξ的表达无明显影响(P>0.05)。结论 PMA慢性处理大鼠血管平滑肌细胞可抑制传统型PKC-α和新型PKC-δ、ε、θ的表达,对非典型PKC-ζ的表达没有影响。
Objective To investigate the changes of protein kinase C （PKC） isoforms in rat vascular smooth muscle cells （VSMCs） treated with prolonged incubation of phorbol-12-myristate-13-acetate （PMA）. Methods Primary rat vascular smooth muscle cells cultured in vitro were divided into blank group, 1, 5, and 10 μmol/L PMA-treated groups, and 0.25%v DMSO-treated group, and all cells were treated for 4 h. The other cells were treated with 10 μmol/L PMA for 1, 4, and 24 h and with 0.25% DMSO for 24 h. The levels of PKCs were measured using Western blotting. Results In 1, 5, and 10 μmol/L PMA-treated groups for 4 h, the levels of PKC-α had no significant changes （P 〉 0.05）, while in 10 μmol/L PMA-treated group for 24 h, PKC-α was down-regulated significantly （P 〈 0.05）. PKC-α expression was down-regulated obviously in VSMCs treated with PMA （〉5 μmol/L for over 4 h）（P 〈 0.01）. PKC-α expression had no changes in PMA- treated groups （〈 5 μmol/L shorter than 4 h） （P 〉 0.05）, while PKC-α decreased remarkably in PMA group （ 10μmol/L for 1 h） （P 〈 0.01）. PKC-0 expression was down-regulated obviously in VSMCs treated with PMA （ 〉 5 μmol/L for over 4 h）（P 〈 0.01）. Level of PKC-μ had no changes in PMA group （10 μmol/L for 24 h）（P 〉 0.05）. Conclusion Prolonged treatment with PMA down-regulated classical PKC-α and novel PKC- δ, ε, and θ in VSMCs, while had no effect on atypical PKC-ζ.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2013年第12期1560-1565,1572,共7页
Journal of Shanghai Jiao tong University:Medical Science
基金
国家自然科学基金(30972842)~~
关键词
佛波酯
血管平滑肌细胞
蛋白激酶C
phorbol-12-myristate-13-acetate
vascular smooth muscle cells
protein kinase C
