Alkylamides from Echinacea are a new class of cannabinomimetics -: Cannabinoid type 2 receptor-dependent and -independent immunomodulatory effects
Alkylamides from Echinacea are a new class of cannabinomimetics -: Cannabinoid type 2 receptor-dependent and -independent immunomodulatory effects
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DOI：
10.1074/jbc.m601074200
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发表时间：
2006-05-19
影响因子：
4.8
通讯作者：
Gertsch, Juerg
中科院分区：
文献类型：
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作者：
Raduner, Stefan;Majewska, Adriana;Gertsch, Juerg
来自紫锥菊的烷基酰胺（alkamides）通过大麻素2型（CB 2）受体调节人单核细胞/巨噬细胞中的肿瘤坏死因子α mRNA表达（Gertsch，J.，Schoop河，昆茨勒，美国，和Suter，A. 04 The Lett.（2004）577，563-569）。在这里，我们表明，烷基酰胺十二碳-2E，4 E，8 Z，10 Z-四烯酸异丁基酰胺（A1）和十二碳-2E，4 E-二烯酸异丁基酰胺（A2）比内源性大麻素更强地结合到CB 2受体。通过置换合成的高亲和力大麻素配体[H-3] CP-55，940来测定A1和A2的Ki值（CB 2类似于60 nM; CB 1> 1500 nM）。分子模拟表明，烷基酰胺结合在溶剂可访问的腔在CB 2，由H-键合和π-π相互作用。在用49种其他与CB 2和CB 1相关的受体进行的筛选中，可以显示A1和A2特异性结合CB 2和CB 1。A1和A2升高CB 2阳性但不升高CB 2阴性早幼粒细胞HL 60细胞中的总细胞内Ca 2+，这种作用被CB 2拮抗剂SR 144528抑制。在50 nM时，A1、A2和内源性大麻素anandamide（CB 2 Ki> 200 nM）以看似CB 2依赖性的方式上调人全血中的组成性白细胞介素（IL）-6表达。A1、A2、大麻素、CB 2拮抗剂SR 144528（Ki < 10 nM）以及非CB 2结合烷基酰胺十一碳-2E-烯、8，10-二炔酸异丁基酰胺均以CB 2非依赖性方式显著抑制脂多糖诱导的肿瘤坏死因子α、IL-1 β和IL-12 p70表达（5-500 nM）。烷基酰胺和大麻素对人全血中抗CD 3刺激的细胞因子表达与抗CD 28刺激的细胞因子表达也显示出弱的差异效应。总体而言，烷基酰胺、花生四烯酸和SR 144528可有效抑制人全血中脂多糖诱导的炎症，并对细胞因子表达产生调节作用，但这些作用并不完全与CB 2结合相关。
Alkylamides (alkamides) from Echinacea modulate tumor necrosis factor alpha mRNA expression in human monocytes/ macrophages via the cannabinoid type 2 (CB2) receptor (Gertsch, J., Schoop, R., Kuenzle, U., and Suter, A. (2004) FEBS Lett. 577, 563-569). Here we show that the alkylamides dodeca-2E,4E,8Z,10Z- tetraenoic acid isobutylamide (A1) and dodeca-2E, 4E-dienoic acid isobutylamide (A2) bind to the CB2 receptor more strongly than the endogenous cannabinoids. The K-i values of A1 and A2 (CB2 similar to 60 nM; CB1 > 1500 nM) were determined by displacement of the synthetic high affinity cannabinoid ligand [H-3] CP-55,940. Molecular modeling suggests that alkylamides bind in the solvent-accessible cavity in CB2, directed by H-bonding and pi-pi interactions. In a screen with 49 other pharmacologically relevant receptors, it could be shown that A1 and A2 specifically bind to CB2 and CB1. A1 and A2 elevated total intracellular Ca2+ in CB2-positive but not in CB2-negative promyelocytic HL60 cells, an effect that was inhibited by the CB2 antagonist SR144528. At 50 nM, A1, A2, and the endogenous cannabinoid anandamide (CB2 K-i > 200 nM) up-regulated constitutive interleukin (IL)-6 expression in human whole blood in a seemingly CB2-dependent manner. A1, A2, anandamide, the CB2 antagonist SR144528 (Ki < 10 nM), and also the non-CB2-binding alkylamide undeca-2E-ene,8,10-diynoic acid isobutylamide all significantly inhibited lipopolysaccharide-induced tumor necrosis factor alpha, IL-1 beta, and IL-12p70 expression (5-500 nM) in a CB2-independent manner. Alkylamides and anandamide also showed weak differential effects on anti-CD3- versus anti-CD28-stimulated cytokine expression in human whole blood. Overall, alkylamides, anandamide, and SR144528 potently inhibited lipopolysaccharide-induced inflammation in human whole blood and exerted modulatory effects on cytokine expression, but these effects are not exclusively related to CB2 binding.