Dirsch Verena M, Keiss Hans-Peter, Vollmar Angelika M
Dept. of Pharmacy, Center of Drug Research, Butenandtstr. 5-13, 81377, Munich, Germany.
Eur J Nutr. 2004 Feb;43(1):55-9. doi: 10.1007/s00394-004-0437-7. Epub 2004 Jan 6.
The transcription factor NF-kappaB is suggested to play a pivotal role in atherogenesis by regulating the expression of proinflammatory gene products such as adhesion molecules. Garlic is consumed as food or dietary supplement in order to benefit from its antiatherogenic effect.
This study addresses the question whether the garlic metabolites diallyldisulfide (DADS) and allylmercaptane (AM) influence the TNF-alpha-induced activation of NF-kappaB and the NF-kappaB-regulated endothelial gene product E-selectin in human umbilical endothelial cells (HUVECs).
A putative effect of DADS and AM on NF-kappaB activity was examined by measuring NF-kappaB DNA binding activity by an electrophoretic mobility shift assay (EMSA) and on the NF-kappaB transactivation activity by a luciferase reporter gene assay. In addition, an effect of DADS and AM on the expression of the NF- kappaB-regulated gene product E-selectin was investigated by flow cytometry.
We show that neither DADS nor AM (up to 100 micro M) inhibit TNF-alpha (1-10 ng/ml)-induced NF-kappaB DNA binding nor NF-kappaB transactivation activity. The TNF-alpha-induced expression of the endothelial adhesion molecule E-selectin was not reduced in response to DADS or AM.
This study suggests that NF-kappaB is not a major target of garlic metabolites such as DADS or AM.
转录因子核因子-κB(NF-κB)被认为通过调节促炎基因产物如黏附分子的表达在动脉粥样硬化形成中起关键作用。大蒜作为食物或膳食补充剂食用,以从其抗动脉粥样硬化作用中获益。
本研究探讨大蒜代谢产物二烯丙基二硫化物(DADS)和烯丙基硫醇(AM)是否影响肿瘤坏死因子-α(TNF-α)诱导的人脐静脉内皮细胞(HUVECs)中NF-κB的激活以及NF-κB调节的内皮基因产物E-选择素。
通过电泳迁移率变动分析(EMSA)测量NF-κB DNA结合活性以及通过荧光素酶报告基因分析测量NF-κB反式激活活性,来检测DADS和AM对NF-κB活性的假定作用。此外,通过流式细胞术研究DADS和AM对NF-κB调节的基因产物E-选择素表达的影响。
我们发现,DADS和AM(高达100μM)均不抑制TNF-α(1-10 ng/ml)诱导的NF-κB DNA结合或NF-κB反式激活活性。TNF-α诱导的内皮黏附分子E-选择素的表达对DADS或AM无反应性降低。
本研究表明,NF-κB不是DADS或AM等大蒜代谢产物的主要作用靶点。