Beermann Silke, Vauth Marcus, Hein Rabea, Seifert Roland, Neumann Detlef
Institute of Pharmacology, Hannover Medical School, Hannover, Germany.
PLoS One. 2014 Sep 22;9(9):e107481. doi: 10.1371/journal.pone.0107481. eCollection 2014.
Histamine (HA) is recognized by its target cells via four G-protein-coupled receptors, referred to as histamine H1-receptor (H1R), H2R, H3R, and H4R. Both H1R and H4R exert pro-inflammatory functions. However, their signal transduction pathways have never been analyzed in a directly comparable manner side by side. Moreover, the analysis of pharmacological properties of the murine orthologs, representing the main targets of pre-clinical research, is very important. Therefore, we engineered recombinant HEK293 cells expressing either mouse (m)H1R or mH4R at similar levels and analyzed HA-induced signalling in these cells. HA induced intracellular calcium mobilization via both mH1R and mH4R, with the mH1R being much more effective. Whereas cAMP accumulation was potentiated via the mH1R, it was reduced via the mH4R. The regulation of both second messengers via the H4R, but not the H1R, was sensitive to pertussis toxin (PTX). The mitogen-activated protein kinases (MAPKs) ERK 1/2 were massively activated downstream of both receptors and demonstrated a functional involvement in HA-induced EGR-1 gene expression. The p38 MAPK was moderately activated via both receptors as well, but was functionally involved in HA-induced EGR-1 gene expression only in H4R-expressing cells. Surprisingly, in this system p38 MAPK activity reduced the HA-induced gene expression. In summary, using this system which allows a direct comparison of mH1R- and mH4R-induced signalling, qualitative and quantitative differences on the levels of second messenger generation and also in terms of p38 MAPK function became evident.
组胺(HA)通过四种G蛋白偶联受体被其靶细胞识别,这些受体被称为组胺H1受体(H1R)、H2R、H3R和H4R。H1R和H4R均发挥促炎功能。然而,它们的信号转导途径从未以直接可比的方式进行过并列分析。此外,对代表临床前研究主要靶点的小鼠直系同源物的药理特性进行分析非常重要。因此,我们构建了重组HEK293细胞,使其以相似水平表达小鼠(m)H1R或mH4R,并分析了这些细胞中HA诱导的信号传导。HA通过mH1R和mH4R均诱导细胞内钙动员,其中mH1R的作用更为有效。虽然cAMP积累通过mH1R得到增强,但通过mH4R则减少。通过H4R而非H1R对这两种第二信使的调节对百日咳毒素(PTX)敏感。丝裂原活化蛋白激酶(MAPK)ERK 1/2在两种受体的下游均被大量激活,并证明在HA诱导的EGR-1基因表达中发挥功能作用。p38 MAPK也通过两种受体被适度激活,但仅在表达H4R的细胞中在功能上参与HA诱导的EGR-1基因表达。令人惊讶的是,在这个系统中p38 MAPK活性降低了HA诱导的基因表达。总之,使用这个能够直接比较mH1R和mH4R诱导信号传导的系统,在第二信使生成水平以及p38 MAPK功能方面的定性和定量差异变得明显。