葛根素对溶血磷脂酰胆碱诱导血管平滑肌细胞基质金属蛋白酶-9表达的影响

Effect of Puerarin on the expression of matrix metalloproteinase-9 induced by lysophosphatidylcholine in vascular smooth muscle cells

目的:观察葛根素对血管平滑肌细胞基质金属蛋白酶-9(MMP-9)表达的影响,探讨葛根素抗动脉粥样硬化(AS)的作用机制。方法:用不同浓度(1.0～10.0mg/L)的溶血磷脂酰胆碱(LPC)孵育体外培养的大鼠胸主动脉平滑肌细胞24h,观察培养上清液中MMP-9含量的变化。不同浓度(10-6～10-4mol/L)的葛根素预先孵育平滑肌细胞4h,然后加入LPC(5.0mg/L)作用24h,用酶联免疫吸附法(ELISA)检测上清液中MMP-9含量,实时荧光定量PCR(real-time PCR)检测平滑肌细胞MMP-9 mRNA的表达,蛋白印迹法(Western blot)检测NF-κB p65蛋白的表达。结果:LPC能明显增加上清液中MMP-9的含量,而预先应用不同浓度的葛根素干预后,MMP-9含量、mRNA表达以及NF-κB p65蛋白的表达水平均明显降低,并且具有剂量依赖性。结论:LPC通过激活NF-κB p65而使平滑肌细胞MMP-9表达增强,而葛根素对上述效应具有抑制作用。

Objective：To observe the effect of Puerarin on the expression of matrix metalloproteinase-9 （MMP-9） induced by lysophosphatidylcholine （LPC） and investigate the anti-atherosclerosis mechanism of Puerarin. Methods：The vascular smooth muscle cells （VSMCs） were incubated with different concentrations of LPC （1.0-10.0 mg/L） for 24 h to detect the level of MMP-9 in supernatants. The VSMCs were treated with LPC （5.0 mg/L） for 24 h following pretreatment with various concentrations of Puerarin （10^-6-10^-4 mol/L） for 4 h. The level and mRNA expression of MMP-9 were determined by enzyme linked immunosorbent assay （ELISA） and real-time quantitative reverse transcription-polymerase chain reaction （real-time RTPCR） respectively. Western blot analysis was used to investigate the activity of nuclear factor kappa B p65 （NFκB p65）. Results：LPC significantly increased the level of MMP-9. The increased level and mRNA expression of MMP-9,and the activity of NF-κB p65 were markedly inhibited by different concentrations of Puerarin in a dose-dependent manner. Conclusion：LPC enhances the expression of MMP-9 by activating NF-κB p65 in VSMCs,while Puerarin is able to inhibit these effects.