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目的:探讨葡萄籽中的原花青素(GSP)对同型半胱氨酸(HCY)诱导的血管平滑肌细胞(VSMC)增殖、迁移的抑制作用及其机制。方法:采用细胞计数及[3H]-TdR检测细胞增殖,用DCFH-DA、Western blotting法检测GSP对VSMC增殖、迁移的抑制作用及分析其相关的分子信号转导通路。结果:HCY浓度依赖性地诱导VSMC的增殖与迁移,1 mmol/L HCY使VSMC的增殖与迁移分别比对照组升高3.9倍及4.1倍(P<0.01);DCFH-DA结果显示,1 mmol/L HCY使VSMC的活性氧(ROS)水平比对照组升高4倍(P<0.01)。而在不同浓度的GSP(5-20 g/L)处理组,HCY诱导VSMC的增殖、迁移以及活性氧水平都被显著抑制(P<0.01)。在GSP 20 g/L处理组,VSMC的增殖、迁移以及活性氧水平都接近空白对照组。与HCY组相比,GSP还显著降低HCY诱导VSMCMCP-1、IL-6及TNF-α的表达水平(P<0.01)。GSP还阻断HCY诱导的NF-κB的激活及显著降低NF-κB的活性(P<0.01)。结论:GSP通过抑制NF-κB的激活而抑制HCY诱导的血管平滑肌细胞增殖、迁移及炎症反应。
Objective: To investigate the inhibitory effect of proanthocyanidins (GSP) from grape seeds on homocysteine (HCY)-induced proliferation and migration of vascular smooth muscle cells (VSMCs) and its mechanism. Methods: The cell proliferation was detected by cell counting and [3H]-TdR. The inhibitory effect of GSP on the proliferation and migration of VSMC was detected by DCFH-DA and Western blotting and the related molecular signal transduction pathways were analyzed. RESULTS: HCY induced the proliferation and migration of VSMCs in a concentration-dependent manner. The proliferation and migration of VSMCs were increased by 3.9-fold and 4.1-fold compared with the control group, respectively, with 1 mmol/L HCY (P<0.01). The results of DCFH-DA showed that 1 mmol/L HCY. /L HCY increased the reactive oxygen species (ROS) level of VSMC by 4 times compared with the control group (P<0.01). However, in different concentrations of GSP (5-20 g/L) treatment group, HCY-induced VSMC proliferation, migration and active oxygen levels were significantly inhibited (P <0.01). In GSP 20 g/L treatment group, the proliferation, migration, and reactive oxygen species of VSMC were similar to those of the blank control group. Compared with HCY group, GSP also significantly decreased the expression of MCP-1, IL-6 and TNF-α in VSMCs induced by HCY (P<0.01). GSP also blocked HCY-induced activation of NF-κB and significantly decreased NF-κB activity (P<0.01). CONCLUSION: GSP inhibits HCY-induced proliferation, migration and inflammatory response of vascular smooth muscle cells by inhibiting the activation of NF-κB.