Effect of Endotoxin Administration in Trypan Blue Pretreated Mice

Abstract

In a previous paper (7) we reported that administration of endotoxin plus lead acetate to mice resulted in marked formation of lipid peroxide, and the activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in the liver were lower than those in mice treated with endotoxin alone. It has been noted that a blockade of the reticuloendothelial system (RES) increases the sensitivity of animals to the lethal effect of endotoxin (12). However, because of its numerous biological effects on cell metabolism, lead acetate, an RES depressor, can be considered to be a specific blocker different from the other agents. The present study was carried out to observe the effect of endotoxin administration under RES depression by trypan blue.Male ddY mice (Shizuoka Experimental Animal Farm Co., Japan), 6 to 7 weeks old and weighing 22 to 25 g, were purchased and maintained in this laboratory for at least one week. Salmonella typhimurium endotoxin (Westphal preparation obtained from Difco, Detroit, MI, U.S.A.) was used throughout this study. This toxin was suspended in normal saline prior to use. A dose of 50 mg/kg or 200 mgjkg of 1.25% trypan blue (Wako Pure Chemical Industries Ltd., Osaka) was injected intravenously into mice 24 hr before endotoxin administration unless otherwise noted. Control mice were injected intraperitoneally with 0.2 ml of a 0.9% saline solution. Mice were fasted after endotoxin administration.Lipid peroxide in liver tissue homogenates (in 8.1 % sodium dodecyl sulfate[SDS] containing 20% acetate buffer, pH 3.5) was determined fluorometrically with thiobarbituric acid (TBA) by a modification (4) of Vagi's method (14). Isozymes of lactate dehydrogenase were estimated electrophoretically according to the method by which NADH is formed in the presence ofphenazine methosulfate and NBT is reduced to a strongly colored formazan using the "LDH Isozyme-Test Kit" (Wako Pure Chemical Industries Ltd., Osaka). Serum lipoprotein was separated by disk-gel electrophoresis in 3.75% polyacrylamide gel performed with a previously stained lipoprotein according to the method of Narayan et al (3). Table 1 shows the percentage of survival after endotoxin injection in trypan blue (TB)-pretreated mice. TB was injected intravenously into mice at doses of 50 mg/kg, 100 mg/kg, and 200 mg/kg. Twenty-four hours after TB injection, 18 mg/kg of endotoxin was administered intraperitoneally to the mice. TB (200 973

Effect of Calcium Ion on Lipid Peroxide Formation in Endotoxemic Mice

“…The major mechanism of detoxification of endotoxin may be considered to be performed by the reticuloendothelial system (RES), particularly Kupffer cells, in the liver. It is well known that ischemia causes functional and structural damage to liver cells (1,3,20).…”

Preventive Effects of a Chinese Herb Medicine (Sho‐saiko‐to) against Lethality after Recombinant Human Tumor Necrosis Factor Administration in Mice

“…Male ddY (Shizuoka Laboratory Animal Center, Hamamatsu, Japan), 4 weeks old and 18 to 20 g, were purchased and maintained in Tohoku College of Pharmacy Experimental Animal Center for at least one week before the experiment. The From our previous reports (1,18), we demonstrated that the major mechanism of endotoxin clearance from blood may be caused by RES, especially by the Kupffer cells in the liver. We assumed that the Chinese medicines enhanced the host defense mechanism by increasing the RES functions.…”

“…TNF and cachectin have severally been reported to cause a shock syndrome similar to endotoxin shock, and it has been suggested that TNF may be the major mediator of endotoxin shock (3,10). From a series in our study on metabolic response in endotoxemia (1,(15)(16)(17)(18)(19)(20), we paid attention to the pharmacological response of Kampo-Sho-saiko-to (14) in order to improve endotoxin shock. The present study was initially carried out to observe the effects of Sho-saiko-to on the lethal and antitumor activities of rhTNF in mice.…”

“…The From our previous reports (1,18), we demonstrated that the major mechanism of endotoxin clearance from blood may be caused by RES, especially by the Kupffer cells in the liver. We assumed that the Chinese medicines enhanced the host defense mechanism by increasing the RES functions.…”

Metabolic Aspects of Endotoxin as a Model of Septic Shock —Approached from Oxidative Stress—

“…筆者らは長年，ショックモデルとしてのエンドト キシン血症の病態生理学の研究に興味を持ってい た．この間，生化学の進歩を基にしたエンドトキシ ン及びショック研究は目覚しく，また，筆者らもエ ンドトキシン血症の病態機構を明らかにするために フリーラジカル産生， [4][5][6][7][8][9] RES 機能， 10,11) リポプロ テイン代謝， [12][13][14] グリコーゲン枯渇機構， 15,16) 細胞 内 Ca 2＋ 動態， [17][18][19] サイトカイン， 20,21) 一酸化窒素 (nitoric oxide, NO)， [22][23][24] 必須微量元素 25,26) そして 漢方方剤小柴胡湯のショック防止効果 ] i in an individual vital cell was shown by computer graphics using the ARGUS 100CA ststem. B: a: molecular weight of protein (140 kDa) of Ca 2＋  ATPase 40) in liver plasma membrane, b: region of endotoxin-induced membrane damage.…”