【摘要】 为观察葛根素诱导兔血管平滑肌细胞凋亡过程中相关基因mRNA的差异表达 ,探讨葛根素促进血管平滑肌细胞凋亡的分子机制。应用葛根素体外诱导兔血管平滑肌细胞凋亡模型和mRNA差异显示技术观察葛根素诱导与正常细胞间基因差异表达 ,将葛根素作用后高表达的cDNA片段克隆、测序 ,与GeneBank进行同源性分析 ,并用32 P标记探针 ,再与不同浓度葛根素作用的血管平滑肌细胞总RNA进行Northern印迹分析。结果发现 ,1、差异显示图谱出现一些仅在葛根素组高表达而在对照组无表达的cDNA片断。2、将其序列与GeneBank进行同源性比较 ,证实其中 3个序列均与兔葡萄糖调节蛋白 94基因高度同源。3、Northern杂交证实兔葡萄糖调节蛋白 94基因在葛根素作用的血管平滑肌细胞中高表达 ,与差异显示图谱一致。4、灰度扫描分析 ,兔葡萄糖调节蛋白 94基因在血管平滑肌细胞中表达量与葛根素浓度成正比。上述结果提示 ,葛根素诱导血管平滑肌细胞凋亡过程中上调了兔葡萄糖调节蛋白 94基因表达 ,该基因在此过程中可能起到一定作用更多还原

【Abstract】 Aim To investigate the effects of one of the flavone puerarin on the apoptosis of the vascular smooth muscle cell (VSMC) and the molecular mechanism of this effect and observe the influence of puerarin on the expression ofdthe relevant genes of apoptosis. Methods By using the method of differential display RT PCR on the puerarin induced rabbit aorta smooth muscle cell (SMC) apoptosis model, the different expressed fragments were recovered, cloned,sequenced and compared with GeneBank databases. Then labeled the probe by 32 P, and observed the gene expression by northern hybridization with the VSMC mRNA stimulated by puerarin in different concentration. Results 1. Some ESTs showed significant higher level of expression in puerarin group than in control group. 2. Three ESTs were isolated from VSMCs stimulated by puerarin, similar to the gene of glucose regulated protein 94 (GRP94) in the rabbit. 3. Northern hybridization showed the high level expression in puerarin treated VSMC. 4. Gray scale scan proved that GRP94 was positive ratio with puerarin concentration. Conclusion The molecular mechanism of the function of promoting apoptosis of VSMC by puerarin perhaps results from promoting the gene expression of the GRP94 of VSMC.更多还原