Laengle Ulrich W, Markstein Rudolf, Cazaubon Cecile, Roman Danielle
Department of Toxicology & Pathology, Novartis Pharma AG, Basel, Switzerland.
Department of Toxicology & Pathology, Novartis Pharma AG, WKL-126.1.16, Klybeckerstr. 141, CH-4057, Basel, Switzerland.
Jpn J Ophthalmol. 2009 Mar;53(2):159-163. doi: 10.1007/s10384-008-0625-8. Epub 2009 Mar 31.
GLC756, a putative antiglaucoma drug with dopamine D(2) agonist and D(1) antagonist properties, significantly decreases tumor necrosis factor alpha (TNF-alpha) levels in lipopolysaccharide (LPS)-induced rats. The present study describes the effects of GLC756 on cellular adenosine 3', 5'-cyclic monophosphate (cAMP) in relation to TNF-alpha production on LPS-stimulated human acute monocytic leukemia cells.
A human peripheral blood acute monocytic leukemia cell line (THP-1) was activated via LPS. THP-1 cells were incubated with GLC756 or betamethasone (positive control) at concentrations of 1, 10, and 30 microM. The TNF-alpha concentration in supernatant and cAMP levels in cellular extract were measured by enzyme-linked immunosorbent assay 0,1, 2.5, 4.5, 7, and 24 h post-activation.
Compared with LPS controls, both GLC756 at 30 muM and betamethasone at > or =1 microM had a significant inhibitory effect on TNF-alpha release from THP-1 cells 2.5 to 24 h post-activation. Parallel to the TNF-alpha decrease, GLC756 induced significant increases of cellular cAMP 2.5 and 7 h post-activation. Betamethasone had no effect on the cellular cAMP level.
Intracellular signaling pathway leading to inhibition of the production of the proinflammatory cytokine TNF-alpha after GLC756 treatment might be mediated through the second messenger cAMP.
GLC756是一种具有多巴胺D(2)激动剂和D(1)拮抗剂特性的潜在抗青光眼药物，可显著降低脂多糖(LPS)诱导的大鼠体内肿瘤坏死因子α(TNF-α)水平。本研究描述了GLC756对LPS刺激的人急性单核细胞白血病细胞中细胞腺苷3',5'-环磷酸(cAMP)的影响及其与TNF-α产生的关系。
通过LPS激活人外周血急性单核细胞白血病细胞系(THP-1)。将THP-1细胞与浓度为1、10和30μM的GLC756或倍他米松(阳性对照)孵育。在激活后0、1、2.5、4.5、7和24小时，通过酶联免疫吸附测定法测量上清液中TNF-α浓度和细胞提取物中cAMP水平。
与LPS对照组相比，30μM的GLC756和≥1μM的倍他米松在激活后2.5至24小时对THP-1细胞释放TNF-α均有显著抑制作用。与TNF-α降低平行，GLC756在激活后2.5和7小时诱导细胞cAMP显著增加。倍他米松对细胞cAMP水平无影响。
GLC756处理后导致促炎细胞因子TNF-α产生受到抑制的细胞内信号通路可能通过第二信使cAMP介导。
