吴茱萸碱抑制大鼠血管平滑肌细胞增殖及对细胞外信号调节激酶-1表达的影响

Inhibition of evodiamine on rat vascular smooth muscle cell proliferation and impact on ERK-1 expression

目的明确吴茱萸碱对血管紧张素Ⅱ(AngⅡ)诱导血管平滑肌细胞(VSMC)增殖的抑制作用及对细胞外信号调节激酶-1(ERK-1)和丝裂原活化蛋白激酶磷酸酶-1(MKP-1)表达的影响。方法贴壁法培养大鼠胸主动脉VSMC,MTT比色法测定吴茱萸碱对AngⅡ诱导大鼠VSMC增殖的抑制作用,Real-time PCR检测VSMC中增殖细胞核抗原(PCNA)、ERK-1和原癌基因c-myc m RNA的表达,Western blot检测VSMC中MKP-1蛋白的表达。结果吴茱萸碱0.1μmol/L和1.0μmol/L明显抑制AngⅡ(1.0μmol/L)诱导VSMC光密度值的增加(P<0.05,P<0.01),抑制AngⅡ上调ERK-1(P<0.01)、PCNA(P<0.05)和c-myc m RNA(P<0.05)的表达,上调MKP-1蛋白的表达(P<0.05,P<0.01)。结论吴茱萸碱明显抑制AngⅡ诱导的VSMC增殖,其机制与下调ERK m RNA、上调MKP-1蛋白的表达有关。

[Objective ] To study the inhibition of evodiamine, a component of Evodia rutaecarpa （Juss.） Benth, on angiotensin Ⅱ （Ang Ⅱ）-induced proliferation of rat vascular smooth muscle cells （VSMC） and the impact on extraeellular signal-regulated kinase-1 （ERK-1） and mitogen-activated protein kinase phosphatase-1 （MKP-1） expressions. [Methods] Sprague Dawley rats VSMC cultured by the tissue explants method were randomly divided into control, model （Ang Ⅱ 1.0μmol/L） and evodiamine （0.01, 0.10 and 1.0 μmol/L） groups. MTT assay was used to detect the effects of evodiamine on VSMC proliferation. The mRNA expressions of ERK-1, proliferating cell nuclear antigen （PCNA） and c-myc and the protein expression of MKP-1 were as- sessed using RT-PCR and western blotting assay, respectively. [Results] Ang Ⅱ significantly induced VSMC proliferation, as evidenced by the increased VSMC absorbance through MTT assay. Evodiamine （0.10μmol/L, 1.0 μmol/L） markedly decreased Ang Ⅱ （1.0 μmol/L）-enhanced VSMC absorbance （P〈0.05, P〈0.01）, suppressed Ang Ⅱ-induced mRNA over-expressions of ERK-1 （P〈0.01）, PCNA （P〈0.05） and c-myc and increased MKP-1 protein expression （P〈0.05, P〈0.01）. [Conclusions] Evodiamine is effective against Ang-Ⅱ induced VSMC proliferation. The mechanisms might be related to the down-regulation of ERK-1 mRNA expression and the up-regulation of MKP-1 protein expression.