通心络对同型半胱氨酸损伤的内皮细胞的基因表达谱的影响

Effect of Tongxinluo on gene expression profiles of endothelial cells treated with homocysteine

目的:探讨通心络对同型半胱氨酸(Hcy)损伤的人脐静脉内皮细胞(HUVECs)的基因表达谱的影响。方法:原代培养的HUVECs,随机分为对照组、Hcy组和通心络组,提取总RNA并纯化,反转录合成荧光分子(Cy3/Cy5)标记的cDNA探针,采用高通量Affymetric人类全基因组寡核苷酸微阵列芯片(含47 000个基因或基因片段)杂交,杂交信号经扫描和数字化处理,通过生物信息学数据分析推测通心络对同型半胱氨酸损伤的HUVECs细胞功能的影响以及可能的信号通路。结果:与对照组相比,Hcy组有4 343个基因表达上调,316个基因表达下调;与Hcy组相比,通心络组有3 409个基因表达上调,121个基因表达下调。这些共同的差异基因生物功能涉及转录因子调控、激酶、细胞骨架与蛋白合成、转运功能、细胞因子、细胞-细胞受体相互作用和细胞黏附因子等。参与的信号通路主要是与血管新生、凋亡、氧化应激、凝血纤溶和炎症等相关的信号通路,如mTOR信号通路、MAPK信号通路和Toll样受体信号通路。结论:通心络能导致同型半胱氨酸损伤内皮细胞基因表达谱的改变,这些基因改变可能参与了细胞凋亡、氧化应激和凝血纤溶等过程。

AIM：To explore the effects of Tongxinluo on the gene expression profiles of homocysteine-treated human umbilical vein endothelial cells（HUVECs）.METHODS：The cell injury model was established by treating the HUVECs with homocysteine.Cultured HUVECs were divided into 3 groups：control group（without any treatment）,homocysteine group（treated with homocysteine at the concentration of 0.5 mol/L） and Tongxinluo group（trated with homocysteine＋Tongxinluo）.The total RNA isolated from the cells with Trizol method was purified and reversely transcribed for synthesizing Cy3/Cy5 labeled cDNA probe.The probe was hybridized with microarray-based Affymetric high throughput gene expression profile（47 000 genes or gene fragments） to screen the differentially expressed genes among the cells with different treatment.The effect of Tongxinluo on the biological function of HUVECs and the possible signal pathway were analyzed.RESULTS：Compared with the control cells,4 659（9.91%） genes were detected to have marked changes with 4 343 up-regulated and 316 down-regulated expression in homocysteine treated cells.Compared with the cells treated with homocysteine,3 530（7.51%） genes were detected to have marked changes with 3 409 up-regulated and 121 down-regulated expression in the cells treated with homocysteine＋Tongxinluo.These genes were involved in transcription activators,cytoskeleton and protein biosynthesis,transport function,cytokines,cell adhesion molecule and metabolism.Signal pathway analysis indicated that Tongxinluo played roles in certain pathways related to vascular proliferation,apoptosis,oxide stress,coagulation,fibrinolysis and inflammation,such as mTOR,MAPK and Toll-like receptor signaling pathways.CONCLUSION：Tongxinluo alters the gene expression profiles of HUVECs injury induced by homocysteine,resulting in the change and modification of cell apoptosis,oxide stress,coagulation and fibrinolysis.