The first observation of O₂⁻ generation at real time in vivo from non‐Kupffer sinusoidal cells in perfused rat liver during acute ethanol intoxication

Abstract

Experimental procedures Animal investigationsThe study protocol was approved by the Committee on Ethics of animal experimentation of the Japan Immunoresearch Laboratories and extra care was taken to avoid animal suffering throughout the study. Male Wistar rats (Charles River, Japan) weighing 250-300 g were fed on standard laboratory animal chow and tap water ad libitum. Before operation, the rats were anaesthetized by i,p. injection of 50 mg/kg sodium pentobarbital and then were given 1000 units/kg sodium heparin by i.v. injection. ReagentsMCLA, a Cypridina luciferin analogue was obtained from Tokyo Kasei Co. Ltd. It was dissolved in sterile saline (pH 7.4) prior to use. The MCLA concentration was based on e430 = 9,600 M -~ .cm -~ [5]. PMA (Sigma) was dissolved in DMSO and then diluted with KrebsHensleit bicarbonate buffer (KHB), pH 7.4. GdC13 (Aldrich) was dissolved in saline (pH 7.4) just before use. Heat inactivated SOD was prepared by treating 50/,tM Cu, Zn SOD (Sigma) in an oven at 120°C for 30 rain.

Ethanol Consumption and Liver Mitochondria Function

“…In earlier studies it was suggested that the Kupffer cells and endothelial cells generated increased concentrations of ROS as a result of acute ethanol exposure [41,42], but little evidence existed for a similar response by hepatocytes. However, Nakano et al [42] recognized that significant increases in ROS occurred that could not be associated with Kupffer cells, although they associated those elevations to endothelial cells.…”

“…However, Nakano et al [42] recognized that significant increases in ROS occurred that could not be associated with Kupffer cells, although they associated those elevations to endothelial cells. It is now well established that hepatocytes also generate significant levels of ROS in the presence of ethanol.…”

“…Several studies, utilizing either perfused liver preparations [41][42][43][44][45] or hepatocytes [26,[46][47][48][49], have provided evidence for increased ROS production which accompanies acute exposure to ethanol. Most of these studies have reported increased ROS production by monitoring for superoxide anion or hydrogen peroxide, but others have measured increased lipid peroxidation after an acute dose of ethanol [50,51].…”

Nonselective cation channels as effectors of free radical–induced rat liver cell necrosis

“…38,39 Gd 3ϩ has been shown to have no short-term effect (Ͻ3 hours) on free-radical production by liver cells exposed to oxidants. 42,43 As predicted from the selectivity of this channel, removal of Na ϩ from the bathing medium mimicked the protective effect of the NSCC blockers (Fig. 3C).…”

Localization of tumor by chemiluminescence probe during photosensitization action

“…FCLA recently developed by Goto and co-workers, is a Cypridinaluciferin analogue and is characterized by emitting light at the longest wavelength (532 nm) among the CL substances known up to now [8][9][10]. Conventional photosensitized reactions of HpD were performed in vitro and in vivo.…”