Evidence for the formation of a novel nitrosothiol from the gaseous mediators nitric oxide and hydrogen sulphide

Abstract

The gaseous mediators hydrogen sulphide (H 2S) and nitric oxide ( NO) are synthesised in the body from l-cysteine and l-arginine, respectively. In the cardiovascular system, NO is an important regulator of vascular tone and its over- or under-production has been linked to a variety of diseases. The physiological significance of H 2S is not yet clear but, like NO, it exhibits vasodilator activity and may play a part in septic and haemorrhagic shock, hypertension, regulation of cardiac contractility, and in inflammation. To date, there have been no reports of a chemical interaction between H 2S and NO. Here we show that incubation of the H 2S donor, sodium hydrosulphide, with a range of NO donors and NO gas in vitro leads to the formation of a nitrosothiol molecule as determined by a combination of techniques; electron paramagnetic resonance, amperometry, and measurement of nitrite. We further show that this nitrosothiol did not induce cGMP accumulation in cultured RAW264.7 cells unless NO was released with Cu 2+. Finally, using liver homogenates from LPS treated rats we present evidence for the endogenous formation of this nitrosothiol. These findings provide the first evidence for the formation of a novel nitrosothiol generated by reaction between H 2S and NO. We propose that generation of this nitrosothiol in the body may regulate the physiological effects of both NO and H 2S.