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Endomorphine 1 and Endomorphine 2 modulates nitric oxide release from mouse macrophages in vitro and in vivo

Introduction: Endomorphine 1 and Endomorphine 2 are novel opioide tetra peptides widely distributed throughout nervous system and highly selective to &micro ; ; ; -opioid receptor. Under inflammatory conditions various types of immune cells produce opioide peptides like enkephalins and endorphins. In addition, macrophages express &micro ; ; ; -opioid receptors indicating the possible involvement of endomorphins in modulation of macrophage inflammatory response and release of reactive radicals, like nitric oxide. The main interest of the present study is the effect of endomorphin 1 and endomorphin 2 on mouse macrophages J774 cell line or peritoneal macrophages on nitric oxide (NO) release. We are measured the release of NO from peritoneal macrophages isolated from mouse treated in vivo with i.p. injection of endomorphine 1 and 2 and challenged in vitro with lipopolisaccharide (LPS). Material and Methods: Nitric oxide in cell supernatants was measured using ISO NO MKII amperometer for real time nitric oxide release measurement, and Griees method for measuring nitrites and nitrates in cell culture supernatants. Results: Results showed that in vitro neither endomorphine 1 nor endomorphine 2 effect NO release in J774 and peritoneal macrophages (Griess method of determination). However, priming or suppressive effect (depending of endomorphine 1 and endomorphine 2 dose) on macrophages challenged with LPS after i.p. injection of endomorphine was noticed. On contrary using real time amperometric method for detection of nitric oxide we noticed that only endomorphine 1 stimulate early activation of iNOS in J774 macrophages in vitro. This effect is inhibited with iNOS specific inhibitor (1400W), while &micro ; ; ; -opioid receptor antagonist (beta-funaltrexamine) did not affect this activation. Conclusion : Endomorphines 1 and 2 differently modulates the release of nitric oxide from peritoneal macrophages treated in vivo with endomorphins, in dose dependent manner. In vitro J774 cell line and peritoneal macrophages did not release NO after stimulation with endomorphin as measured with Griess method. However early activation of iNOS with endomorphin 1 was demonstrated using real time measurement amperometric method.

Endomorphine 1 and Endomorphine 2; nitric oxide

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