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[目的]观察肌细胞增强因子2A(Myocyte enhancer factor 2A、MEF2A)在肝星状细胞(Hepatic StellateCell、HSC)活化过程中的可能作用及四甲基吡嗪对其影响,进一歩探讨四甲基吡嗪抗肝纤维化的作用机制。[方法]采用SD大鼠肝脏原位灌流消化法获得HSC,并原代及传代培养,分离的HSC在塑料细胞培养皿中培养后逐步活化,分别收集新分离(0d)和经培养(1、2、3、4、5、6、7、8d)的HSC,观察MEF2A与HSC活化指标α-平滑肌肌动蛋白(α-smooth muscle action、α-SMA)表达的关系;取传2～4代HSC,加四甲基吡嗪至终浓度50、100、150、200mg/L,分组培养48h,观察四甲基吡嗪对HSC增殖抑制作用的同时,对MEF2A基因及蛋白表达的影响。[结果]①随着HSC培养逐步活化,细胞内MEF2A及α-SMA逐步增加,二者呈正相关(r=0.90,P<0.05),且MEF2ADNA结合活性同步增强。②随着四甲基吡嗪浓度增加,对HSC增殖的抑制作用增强,同时对MEF2A表达抑制作用逐渐增强。[结论]MEF2A可能参与了HSC活化过程,四甲基吡嗪抑制HSC增殖作用可能与有效抑制MEF2A表达有关。
[Objective] To observe the possible role of Myocyte enhancer factor 2A (MEF2A) in the activation of hepatic stellate cells (HSC) and the influence of tetramethylpyrazine on it, Pyrazine anti-liver fibrosis mechanism of action. [Method] HSCs were harvested from the liver of SD rats by in situ perfusion and digestion. Primary and subculture HSCs were separated and cultured in plastic cell culture dishes. HSCs were collected and harvested separately (0, 2, 3, 4, 5, 6, 7, 8d) HSCs. The relationship between MEF2A and the expression of α-smooth muscle action (α-SMA) HSC plus tetramethylpyrazine to the final concentration of 50, 100, 150, 200 mg / L and cultured for 48 hours. The effects of tetramethylpyrazine on the proliferation of HSC were observed, and the effect on MEF2A gene and protein expression was also observed. [Results] ① With the gradual activation of HSC culture, the intracellular MEF2A and α-SMA gradually increased, the two were positively correlated (r = 0.90, P <0.05), and MEF2ADNA binding activity increased synchronously. ② As the concentration of tetramethylpyrazine increased, the inhibitory effect on HSC proliferation increased, while the inhibitory effect on MEF2A gradually increased. [Conclusion] MEF2A may be involved in the activation of HSC. The inhibitory effect of tetramethylpyrazine on the proliferation of HSC may be related to the inhibition of MEF2A expression.