15-KETE对大鼠肺动脉平滑肌细胞上ERK1／2活性的影响

目的从细胞分子水平上研究15-酮基二十碳四烯酸（15-ketoeicosatetretraenoicacid，15-KETE）对大鼠肺动脉平滑肌细胞上ERKl／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（pulmonaryarterialsmoothcells，PASMCs），使用Westernblot技术观察15-KETE对大鼠PASMCs上ERKl／2活性的影响。结果1）10～mol／L15-KETE作用从0．5h到24h与对照组（未加15-KETE）比较，对ERKl／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、8．0h明显增强P—ERKl／2（P〈0．05）。3）与对照组比较，10、10^-7和10～mol／L15-KETE明显增强p-ERKl／2（P〈0．05），随15-KETE浓度增大，p-ERKl／2增多。4）与15-KETE作用比较，ERKI／2抑制剂PD9805920μmol／L、50txmol／L和U01260．1txmol／L、1btmol／L均能抑制15-KETE对ERKl／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ERKl／2表达没有明显的影响，但能激活肺动脉血管平滑肌细胞ERKI／2．使其磷酸化，并呈时间一剂量依赖关系。ERKl／2通路的特异性抑制剂PD98059、U0126能抑制15-KETE对ERKl／2的磷酸化。

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

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.