Substance P‐induced cyclooxygenase‐2 expression in human umbilical vein endothelial cells

Substance P (SP) is a neuropeptide involved in neurogenic inflammation and an agonist for NK1, NK2, and NK3 receptors. SP induces prostaglandin (PG) production in various cell types, and these eicosanoids are responsible for numerous inflammatory and vascular effects. Cyclooxygenase (COX) are needed to convert arachidonic acid to PGs. The study evaluated the effect of SP on COX expression in human umbilical vein endothelial cells (HUVEC). COX‐2 protein expression was upregulated by SP with a peak at 100 nM and at 20 h; in the same experimental conditions COX‐1 protein expression was unchanged. A correlation between COX‐2 expression and PGI2 and PGE2 release was detected. Dexamethasone (DEX) inhibited SP‐mediated COX‐2 expression. Mitogen‐activated protein kinases (MAPK) p38 and p42/44 were activated by SP, whereas SB202190 and PD98059, inhibitors of these kinases, blocked COX‐2 expression. 5,5‐dimethyl‐3‐(3‐fluorophenyl)‐4‐(4‐methylsulphonyl)phenyl‐2(5H)‐furanone (DFU), an experimental selective COX‐2 inhibitor, blocked SP‐induced PG release. By RT–PCR and Western blot analysis, we demonstrated that NK1 and NK2 but not NK3 receptors are present on HUVEC. Selective NK1 and NK2 agonists, namely [Sar9, Met(O2)11]SP and [β‐Ala8] NKA(4–10), upregulated COX‐2 protein expression and PG production, whereas senktide (Suc–Asp–Phe–MePhe–Gly–Leu–Met–NH2), a selective NK3 agonist, was ineffective in this respect. The NK1 selective antagonist L703,606 ((cis)‐2‐(diphenylmethyl)‐N‐((2‐iodophenyl)‐methyl)‐1‐azabicyclo(2.2.2)octan‐3‐amine) and the NK2 selective antagonist SR 48,968 ((S)‐N‐methyl‐N‐(4‐(4‐acetylamino‐4‐phenylpiperidino)‐2‐(3,4 dichlorophenyl)butyl) benzamide) competitively antagonised SP‐induced effects. The study shows HUVEC to possess functional NK1 and NK2 receptors, which mediate the ability of SP to induce expression of COX‐2 in HUVEC, thus showing a previously‐undetected effect of SP on endothelial cells. British Journal of Pharmacology (2006) 147, 681–689. doi:10.1038/sj.bjp.0706660