Sjögren's syndrome, an inflammatory disease affecting the lacrimal and salivary glands, is the leading cause of aqueous tear-deficient type of dry eye. We previously showed that interleukin-1beta (IL-1beta) protein is up regulated in the lacrimal gland of a murine model of Sjögren's syndrome and that exogenous addition of this cytokine inhibits neurotransmitter release and lacrimal gland protein secretion. In the present study we investigated the role of c-Jun NH2-terminal kinase (JNK) in IL-1beta-mediated inhibition of lacrimal gland secretion and tear production. In vitro, IL-1beta induced a time-dependent activation of JNK with a maximum 7.5-fold at 30 min. SP600125, a JNK inhibitor, inhibited, in a concentration-dependent manner, IL-1beta-induced activation of JNK with a maximum of 87% at 10(-4) m. In vivo, IL-1beta stimulated JNK and the expression of the inducible isoform of nitric oxide synthase (iNOS). IL-1beta inhibited high KCl and adrenergic agonist induced protein secretion by 85% and 66%, respectively. SP600125 alleviated the inhibitory effect of IL-1beta on KCl- and agonist-induced protein secretion by 79% and 47%, respectively, and completely blocked the expression of iNOS. Treatment for 7 days with SP600125 increased tear production in a murine model of Sjögren's syndrome dry eye. We conclude that JNK plays a pivotal role in IL-1beta-mediated inhibition of lacrimal gland secretion and subsequent dry eye.