ERK参与调节人脐动脉平滑肌细胞内SMAD2/3蛋白及其mRNA的表达

ERK regulates the expression of SMAD2/3 protein and mRNA in human umbilical artery smooth muscle cells

目的了解ERK信号转导通路对人脐动脉平滑肌细胞(hUASMC)内SMAD2/3蛋白及其mRNA表达的影响。方法原代培养hUASMC,实验分四组:1)对照组,2)PDGF组,3)ERK阻断剂组,4)PDGF+ERK阻断剂组。分别用免疫细胞化学和RT-PCR法测hUASMC内磷酸化SMAD2/3蛋白和SMAD2/3mRNA的表达。结果1)与对照组相比,PDGF组hUASMC细胞内磷酸化SMAD2/3蛋白的表达增强(P<0·01);2)四组hUASMC细胞内SMAD2/3mRNA的表达无明显差异。结论在hUASMC中,ERK通路可促进SMAD2/3蛋白的磷酸化,但对SMAD2/3mRNA的表达没有影响。

Objective To investigate the effects of extracelluar signal regulated kinase （ERK） pathway on the expression of SMAD2/3 protein and mRNA in human umbilical artery smooth muscle cells （hUASMC）. Methods hUASMC were primary cultured in vitro ,then the hUASMC were divided into four groups ： 1 ） control group ,2 ） PDGF group,3） ERK blocking agent group, 4） PDGF ＋ ERK blocking agent group. The expression of phosphorylated SMAD2/3 （p-SMAD2/3） protein in hUSMC were detected by inmunohistoehemistry method and the expression of SMAD2/3 mRNA were detected by RT-PCR technology. Results 1 ） The expression of p-SMAD2/3 protein in hUSMC of PDGF group was higher than that of control（P 〈0.01 ） ; 2）There was no difference on the expression of SMAD2/3 mRNA in hUSMC between the four groups. Conclusion The ERK pathway could promote the expression of p-SMAD2/3 protein and have no effection of the expression of SMAD2/3 mRNA in hUASMC.