15-KETE对大鼠肺动脉平滑肌细胞上ERK1/2活性的影响  

Effect of 15-KETE on ERK1/2 Activity in Rat Pulmonary Artery Smooth Muscle Cells

作　　者：郭守利[1] 张一飞[2] 郝天军[1] 孙艳[3] 

机构地区：[1]哈尔滨医科大学第二临床医学院实验动物中心,哈尔滨150086 [2]哈尔滨医科大学第二临床医学院药学部,哈尔滨150086 [3]哈尔滨医科大学公共卫生学院职业卫生与职业医学教研室,哈尔滨150086

出　　处：《实验动物科学》2013年第6期26-30,40,共6页Laboratory Animal Science

基　　金：黑龙江省应用技术研究与开发计划项目(No.PC13S011)

摘　　要：目的从细胞分子水平上研究15-酮基二十碳四烯酸(15-ketoeicosatetretraenoic acid,15-KETE)对大鼠肺动脉平滑肌细胞上ERK1/2的活性的影响。方法通过酶法分离、培养SD大鼠肺动脉平滑肌细胞(pulmonary arterial smooth cells,PASMCs),使用Western blot技术观察15-KETE对大鼠PASMCs上ERK1/2活性的影响。结果 1)10-6mol/L 15-KETE作用从0.5 h到24 h与对照组(未加15-KETE)比较,对ERK1/2总量没有明显的影响。2)与对照组比较,15-KETE处理0.5、1.0、2.0、4.0、8.0 h明显增强p-ERK1/2(P<0.05)。3)与对照组比较,10-8、10-7和10-6mol/L 15-KETE明显增强p-ERK1/2(P<0.05),随15-KETE浓度增大,p-ERK1/2增多。4)与15-KETE作用比较,ERK1/2抑制剂PD98059 20μmol/L、50μmol/L和U0126 0.1μmol/L、1μmol/L均能抑制15-KETE对ERK1/2的活化(P<0.05),并且随着抑制剂浓度增大,抑制作用加强。结论 15-KETE对大鼠肺动脉血管平滑肌细胞上的ERK1/2表达没有明显的影响,但能激活肺动脉血管平滑肌细胞ERK1/2,使其磷酸化,并呈时间-剂量依赖关系。ERK1/2通路的特异性抑制剂PD98059、U0126能抑制15-KETE对ERK1/2的磷酸化。Objective To investigate rat pulmonary artery smooth muscle collagen plus elastase digestion, and the effect of 15-ketoeicosatetretraenoic acid（15-KETE） on ERK1/2 cells （PASMCs）. Method PASMCs of SD rats were collected activity m by type I cultured in control and defined media. The effect of 15-KETE on ERK1/2 activity in rat pulmonary artery smooth muscle cells was investigated by Western blot. Result 15-KETE had no effect on ERK1/2＇ s expression and activity at 10 - 6 mol/L from 0.5 h to 24 h. But it could increase the expression of p-ERK1/2 after 0.5, 1, 2, 4 and 8 h incubation （P 〈 0.05） ; 15-KETE, at concentration of 10-s 10-6 mol/L, could significantly increase the expression of p- ERK1/2 （P 〈 0.05） ; PD98059 and U0126, two specific inhibitors of ERK1/2, could significantly inhibit the effect of 15-KETE （at concentration of 10-6 mol/L） on ERK1/2 activity （P 〈 0.05）, when PD98059 was added at 20 and 50 ~mol/L and U0126 at 0. 1 and 1 p^mol/L respectively. Conclusion 15-KETE has no effect on ERK1/2 expression, but can activate ERK1/2 in PASMCs and induce its phosphorylation in a time- and concentration-dependent manner. PD98059 and U0126 can inhibit the effect of 15 - HETE on ERKI/2 phosphorylation.

关 键 词：15-KETE 细胞外信号调节激酶 肺动脉血管平滑肌细胞 

分 类 号：R332[医药卫生—人体生理学]