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本文旨在观察血管紧张素II (angiotensin II, Ang II)对血管平滑肌细胞核转录因子-κB (nuclear factor-κB, NF-κB)的活性及骨形成蛋白-2 (bone morphogenetic protein-2, BMP-2)表达的影响,以探讨Ang II参与动脉粥样硬化的机制,并探讨川芎嗪是否能抑制Ang II的促动脉粥样硬化作用。采用Western blot、免疫组化和原位杂交等方法分别检测Ang II刺激和川芎嗪干预后NF-κB活性、BMP-2 蛋白和 mRNA 表达的变化。结果显示:(1) Ang II 刺激激活 NF-κB。Ang II 刺激 15 min 即有 NF-κB p65核转移,30 min 达高峰(P<0.01),1 h 后减退。川芎嗪抑制 Ang II诱导的NF-κB激活,与Ang II组比较,川芎嗪 + Ang II组NF-κB活性显著降低 (P<0.01)。 (2) Ang II 刺激 6 h 时 BMP-2 表达增强(P<0.05),12 h 时减弱(P<0.01),24 h 时更弱(P<0.01)。川芎嗪 + Ang II组中,川芎嗪干预6 h 时BMP-2 表达亦增强,12 与24 h 时保持正常水平。 (3) 川芎嗪对正常细胞的NF-κB活性和BMP-2 表达无影响。以上结果表明,Ang II刺激后激活NF-κB并最终使生长抑制因子BMP-2 表达下降,这可能是其参与动脉粥样硬化发生的机制之一。BMP-2 一过性增高可能不依赖NF-κB通路的激活。川芎嗪可抑制Ang II诱导的NF-κB激活与 BMP-2 表达降低,提示它在抗动脉粥样硬化形成中起重要作用。
The purpose of this study was to observe the activity of angiotensin II (Ang II) on nuclear factor-κB (NF-κB) and bone morphogenetic protein-2 (BMP-) in vascular smooth muscle cells. 2) The effect of expression to explore the mechanism of Ang II involvement in atherosclerosis, and to explore whether tetramethylpyrazine can inhibit Ang II’s atherosclerosis. Western blot, immunohistochemistry and in situ hybridization were used to detect the changes of NF-κB activity, BMP-2 protein and mRNA expression after Ang II stimulation and Ligustrazine treatment. The results showed that: (1) Ang II stimulated the activation of NF-κB. The NF-κB p65 nuclear transfer occurred 15 min after Ang II stimulation and peaked at 30 min (P<0.01), and decreased after 1 h. Ligustrazine inhibited the activation of NF-κB induced by Ang II. Compared with Ang II group, the activity of NF-κB in Ligustrazine + Ang II group was significantly decreased (P<0.01). (2) BMP-2 expression was increased at 6 h after Ang II stimulation (P<0.05), decreased at 12 h (P<0.01), and was weaker at 24 h (P<0.01). In the tetramethylpyrazine + Ang II group, the expression of BMP-2 was increased at the 6th hour after the intervention of tetramethylpyrazine, and it remained at the normal level at 12 and 24 hours. (3) Ligustrazine has no effect on the activity of NF-κB and BMP-2 in normal cells. These results indicate that Ang II stimulates NF-κB activation and ultimately decreases the expression of the growth inhibitory factor BMP-2, which may be one of the mechanisms involved in atherosclerosis. Transient increase in BMP-2 may not depend on activation of the NF-κB pathway. Ligustrazine can inhibit Ang II-induced activation of NF-κB and decrease of BMP-2 expression, suggesting that it plays an important role in the development of anti-atherosclerosis.