Role of Zn(2+) in oxidative stress caused by endotoxin challenge.

The role of Zn(2+) in oxidative stress during endotoxemia was investigated. In rats fed a Zn(2+)-deficient diet (Zn(2+) concentration of less than 1.5 mg/kg) for 8 weeks, the Zn(2+) level in the serum was about 62% lower than that in rats fed a Zn(2+)-adequate diet (Zn(2+) concentration, 50 mg/kg). The Zn(2+) level in serum 18 h after administration of endotoxin (6 mg/kg, i.p.) to Zn(2+)-deficient diet rats was markedly lower than that of the endotoxin/Zn(2+)-adequate diet group. Lipid peroxide formation in the liver of Zn(2+)-deficient diet rats was markedly increased 18 h after endotoxin injection compared with that in the endotoxin/Zn(2+)-adequate diet group. Metallothionein in the liver of endotoxin/Zn(2+)-adequate diet rats was increased more than 17-fold by endotoxin administration, while a markedly lower level of metallothionein was observed in the endotoxin/Zn(2+)-deficient diet group. On the other hand, treatment with ZnSO(4) (100 microM) significantly increased endotoxin (1 microg/ml)-induced tumor necrosis factor-alpha (TNF-alpha) production in J774A.1 cells. Our results clearly demonstrated that treatment with ZnSO(4) significantly inhibited the endotoxin-induced increase in intracellular Ca(2+) level in J774A.1 cells. However, a cell membrane-permeable Zn(2+) chelator, N,N,N',N'-tetrakis (2-pyridylmethyl) ethylenediamine (TPEN, 1 microM), did not affect the endotoxin-induced TNF-alpha production or Ca(2+) level in J774A.1 cells. In addition, we investigated whether Zn(2+) can suppress nitric oxide (NO) generation and cytotoxicity in endotoxin-treated cells. Treatment with ZnSO(4) (50 microM) significantly inhibited endotoxin-induced NO production in J774A.1 cells, but did not affect endotoxin-induced cytotoxicity. These findings suggest that zinc may play an important role, at least in part, in the oxidative stress during endotoxemia.

Preventive effects of a verapamil against tumor necrosis factor-alpha-induced shock symptoms: approached from lipoprotein metabolic disorders.

We examined the role of intracellular Ca2+ in the mechanism of the preventive effects of the Ca2+-channel blocker verapamil against lipoprotein disturbances during tumor necrosis factor (TNFa)-induced shock syndrome. The heparin-releasable lipoprotein lipase (LPL) activity in plasma of TNFalpha (5 X 10(4) units/mouse, i.v.)-injected mice was markedly lower at 4 h post-intoxication than in the controls. In mice treated with verapamil (10 mg/kg, s.c.), the activity of LPL 4 h after TNFalpha injection was significantly higher than in mice treated with TNFalpha alone. On the other hand, on polyacrylamide gel disk electrophoresis, very low density lipoprotein (VLDL) and high density lipoprotein (HDL) fractions in the sera of TNFalpha-injected mice were increased and reduced, respectively, relative to the controls. The administration of verapamil clearly prevented the lipoprotein damage arising from TNFalpha challenge. We investigated whether verapamil could suppress TNFalpha generation in endotoxin-treated J774A.1 cells. Treatment with verapamil (30 microM) markedly inhibited endotoxin (1 microg/ml)-induced TNFalpha production in these cells. These findings suggest that the concentration of intracellular Ca2+ may contribute to the extent of lipoprotein disturbances in plasma, which results from LPL suppression in TNFalpha-induced shock syndrome. Verapamil may, therefore, protect against some of the various disturbances caused by changes in Ca2+ mobilization through its ability to inhibit TNFalpha production in septic shock.

Significant improvement of insulin resistance of GK rats by treatment with sodium selenate.

We studied the effect of sodium selenate on insulin resistance of Goto-Kakizaki (GK) rats. Rats were kept on standard laboratory chow with and without i.p. injections of sodium selenate (0.173 mg/kg body weight) for 14 days, and then subjected to the glucose clamp. The glucose clamp studies confirmed an improvement in insulin-stimulated glucose disposal in GK rats treated with sodium selenate, with respect to both insulin sensitivity and responsiveness. This amelioration of insulin resistance may be partly due to a direct effect of the sodium selenate on peripheral tissues. 2-Deoxyglucose uptake in sodium selenate-treated adipocytes was increased and the insulin findings suggest that sodium selenate increases not only insulin sensitivity but also insulin responsiveness. Sodium selenate also accelerated glucose incorporation into adipocytes of rats, suggesting that the action of sodium selenate is peripheral. Interestingly, sodium selenate at a high concentration (about 1 mmol/L) was more effective than insulin in enhancing glucose uptake. The present study suggested that sodium selenate treatment led to substantial improvement in peripheral insulin resistance.

Preventive effects of a traditional Chinese medicine (Sho-saiko-to) on septic shock symptoms; approached from heme metabolic disorders in endotoxemia.

Sho-saiko-to, one of the the most frequently prescribed Kampo medicines, is used to treat chronic hepatitis and has shown confirmed clinical efficacy. The present study was performed with respect to heme metabolism to study the preventive effects of Sho-saiko-to against endotoxemia. Endotoxin was injected intraperitoneally at a dose of 6 mg/kg into Sho-saiko-to (500 mg/kg/d, p.o.)-pretreated rats, and its administration clearly prevented the endotoxin-induced hypoferremia. In rats pretreated with Sho-saiko-to, the activity of hepatic delta-aminolevulinate synthetase and cytochrome P-450 level 18 h after endotoxin injection were significantly increased as compared to rats treated with endotoxin alone. Similarly, Sho-saiko-to significantly depressed the endotoxin-induced increase in heme oxygenase activity in liver microsomes. These findings suggested that the extent of shock syndrome caused by endotoxin may be due, at least in part, to changes in heme metabolic disturbance during endotoxemia. Sho-saiko-to may therefore protect rats against lethality caused by endotoxin through its ability to regulate the heme metabolism in septic shock.