Lipid peroxidation and total glutathione after different intensities of resistance exercise in trained men.

AIM: Since studies on resistance exercise and oxidative stress markers show contradictory results, it is not clear whether different intensities of exercise are the determinant of changes in such markers. The objective of this study was to investigate the acute effects of different intensities of resistance exercise on lipid peroxidation and total glutathione in previously resistance trained men. METHODS: Eight male subjects with at least 2 years of resistance training experience performed two different resistance exercise protocols: low-intensity (LI), 60% of one repetition maximum (1RM) and high-intensity (HI), 85% of 1RM. Both protocols involved seven exercises and subjects performed one set of each exercise. Blood samples were obtained before and immediately after exercise for lipid peroxidation and total glutathione analysis. RESULTS: The results indicated a significant difference in total workload (load multiplied by repetitions performed) between the LI and HI protocols (P<0.05) and no differences on lipid peroxidation and total glutathione after both LI and HI protocols. CONCLUSION: This study suggests that resistance exercise protocols composed of a single set of seven exercises, regardless of the intensity or total workload do not induce to oxidative stress, suggesting that volume is the main variable to induce oxidative stress in previously resistance trained individuals.

Evidence that macrophages transfer arachidonic acid and cholesterol to tissues in vivo.

Our previous studies have shown that [(14)C]-labelled cholesterol (CHOL) and arachidonic acid (AA) are transferred from macrophages (Mphi) to lymphocytes (LY) when these cells are co-cultured. In this study, we investigated whether these lipids can be transferred from control and thioglycollate-elicited Mphi (THIO-elicited Mphi) to various tissues and organs in vivo. For this purpose, control and THIO-elicited Mphi were pre-treated with [(14)C]-AA and [(3)H]-CHOL and then injected into the jugular vein of adult rats. More than 75% of the radioactivity injected was found in the liver of rats treated with [(14)C]-AA labelled-Mphi either control and THIO-stimulated. The radioactivity of [(3)H]-CHOL labelled Mphi was transferred mainly to the liver (51% in the control Mphi and 23% in the thioglycollate Mphi7) but it was also found in the kidney, lung and spleen. These results support the proposition that the transfer of lipids between cells also occurs in vivo. The full significance of this phenomenon however remains to be elucidated.

Antiproliferative prostaglandins and the MRP/GS-X pump role in cancer immunosuppression and insight into new strategies in cancer gene therapy.

A dramatic complication in late-stage cancer patients is host immunosuppression. Cyclopentenone prostaglandins (CP-PGs) overproduced in cancer may impair the function of the immune system. These agents, if produced at high concentrations, are powerful cytostatic and cytotoxic compounds that may arrest cell proliferation and immune response in cancer. Lymphoid tissues of tumor-bearing animals accumulate large amounts of CP-PGs, whereas the tumor tissue does not. This may be because cancer cells are able to overexpress multidrug resistance-associated protein (Mg(2+)-dependent vanadate-sensitive GS-conjugate export ATPase, MRP/GS-X pump), which extrudes CP-PGs to the extracellular space as glutathione S-conjugates. In contrast, MRP/GS-X pump activity is disproportionately low in lymphocytes. This led us to propose the transfection of lymphocytes with multidrug resistance-associated protein genes (MRP) for further autologous transfusion or direct in vivo delivery to lymphocytes by using adenovirus-retrovirus chimeras in order to restore immune system function in cancer, at least partially. We are currently evaluating MRP-transfected lymphocyte (MTL) therapy, using Walker 256 tumor-bearing rats as a model.

Oxidative stress in the latissimus dorsi muscle of diabetic rats

The purpose of the present study was to investigate the effects of experimental diabetes on the oxidant and antioxidant status of latissimus dorsi (LD) muscles of male Wistar rats (220 +/- 5 g, N = 11). Short-term (5 days) diabetes was induced by a single injection of streptozotocin (STZ, 50 mg/kg, iv; glycemia >300 mg/dl). LD muscle of STZ-diabetic rats presented higher levels of thiobarbituric acid reactive substances (TBARS) and chemiluminescence (0.36 +/- 0.02 nmol/mg protein and 14706 +/- 1581 cps/mg protein) than LD muscle of normal rats (0.23 +/- 0.04 nmol/mg protein and 7389 +/- 1355 cps/mg protein). Diabetes induced a 92 percent increase in catalase and a 27 percent increase in glutathione S-transferase activities in LD muscle. Glutathione peroxidase activity was reduced (58 percent) in STZ-diabetic rats and superoxide dismutase activity was similar in LD muscle of both groups. A positive correlation was obtained between catalase activity and the oxidative stress of LD, as evaluated in terms of TBARS (r = 0.78) and by chemiluminescence (r = 0.89). Catalase activity also correlated inversely with glutathione peroxidase activity (r = 0.79). These data suggest that an increased oxidative stress in LD muscle of diabetic rats may be related to skeletal muscle myopathy

Metabolic response of macrophages to injury promoted by the activated complement system.

In nucleated cells, the swelling promoted by a complement system (CS) attack is not enough to promote cell death, because unlike erythrocytes these cells are able to eliminate cytolytic complement channels from the plasma membrane, by processes that include endocytosis. Several studies have demonstrated that the resistance of nucleated cells to the injury promoted by the CS is related to the cellular metabolism. Despite this, to the present day, no study has clearly related cell survival capacity to injury by the CS to its energetic metabolic status. In macrophages, the challenge imposed by the CS provoked an increase in the total amount of glucose incorporated into fatty acids, including phospholipids and cholesterol; substrates for membrane synthesis. The inhibition of cholesterol synthesis promoted an increase of the cell death rate. These data support the importance of cholesterol metabolism for macrophage resistance to necrosis induced by the activated complement system.

Effects of prostaglandins and nitric oxide on rat macrophage lipid metabolism in culture: implications for arterial wall-leukocyte interplay in atherosclerosis.

Macrophages/foam cells have a pivotal role in atherogenesis although little is known about the way lipid imbalance, a hallmark of atherosclerosis, leads to lipid accumulation in these cells. Modified low-density lipoproteins are associated with macrophage lipid dysfunction in atherosclerosis, but a possible role for altered lipogenesis leading to lipid accumulation remains to be elucidated. Since endothelium-derived nitric oxide (NO) and prostaglandins (PGs) are physiological autacoids whose production may be impaired in atherosclerosis, the effects of these mediators on de novo lipid synthesis in 24-h cultured rat peritoneal macrophages is investigated. In resident (unstimulated) cells, 1 microM PGE2 and the stable analog of PGI2 carbaprostacyclin (cPGI2, 1 microM) deviated the overall [1-14C]acetate from incorporation into cholesterol, free fatty acids and triacylglycerols favoring the formation of phospholipids. In inflammatory (thioglycollate-elicited) macrophages, these eicosanoids likewise reduced 14C-incorporations into all the lipid fractions tested. Also, cPGI2 and PGE2 reduced [4-14C]cholesterol uptake from inflammatory cells but did not interfere in 14C-cholesterol export. The PGE2-derivative PGA2 (10-20 microM) reduced 14C-incorporations into all the lipids in resident cells while it enhanced phospholipid synthesis by up to 129% at the expense of reduced incorporations into the other test lipids. The NO donor S-nitroso-N-acetylpenicillamine (SNAP, 1-10 microM), when added to macrophages in the presence of superoxide dismutase (SOD, to avoid the reaction of superoxide with NO), significantly reduced lipogenesis especially in inflammatory cells. These findings suggest that endothelium-derived NO and PGs may be associated with macrophage lipid accumulation by modulating lipogenesis and cholesterol uptake within these cells.

Effect of trolox C on cardiac contracture induced by hydrogen peroxide.

Hydrogen peroxide (H2O2) perfused into the aorta of the isolated rat heart induces a positive inotropic effect, with cardiac arrhythmia such as extrasystolic potentiation or cardiac contractures, depending on the dose. The last effect is similar to the "stone heart" observed in reperfusion injury and may be ascribed to lipoperoxidation (LPO) of the membrane lipids, to protein damage, to reduction of the ATP level, to enzymatic alterations and to cardioactive compounds liberated by LPO. These effects may result in calcium overload of the cardiac fibers and contracture ("stone heart"). Hearts from male Wistar rats (300-350 g) were perfused at 31 degrees C with Tyrode, 0.2 mM trolox C, 256 mM H2O2 or trolox C + H2O2. Cardiac contractures (baseline elevation of the myograms obtained) were observed when hearts were perfused with H2O2 (Tyrode: 5.9 +/- 3.2; H2O2: 60.5 +/- 13.9% of the initial value); perfusion with H2O2 increased the LPO of rat heart homogenates measured by chemiluminescence (Tyrode: 3,199 +/- 259; H2O2: 5,304 +/- 133 cps mg protein-1 60 min-1), oxygen uptake (Tyrode: 0.44 +/- 0.1; H2O2: 3.2 +/- 0.8 nmol min-1 mg protein-1) and malonaldehyde (TBARS) formation (Tyrode: 0.12 +/- 0; H2O2: 0.37 +/- 0.1 nmol/ml). Previous perfusion with 0.2 mM trolox C reduced the LPO (chemiluminescence: 4,098 +/- 531), oxygen uptake (0.51 +/- 0) and TBARS (0.13 +/- 0) but did not prevent the H2O2-induced contractures (33.3 +/- 16%). ATP (Tyrode: 2.84 +/- 0; H2O2: 0.57 +/- 0) and glycogen levels (Tyrode: 0.46 +/- 0; H2O2: 0.26 +/- 0) were reduced by H2O2. Trolox did not prevent these effects (ATP: 0.84 +/- 0 and glycogen: 0.27 +/- 0). Trolox C is known to be more effective than alpha-tocopherol or gamma-tocopherol in reducing LPO though it lacks the phytol portion of vitamin E to be fixed to the cell membranes. Trolox C, unlike vitamin A, did not prevent the glycogen reduction induced by H2O2. Trolox C induced a positive chronotropic effect that resulted in higher energy consumption. The reduction of energy level seemed to be more important than LPO in the mechanism of H2O2-induced contracture.

Insulin and prostaglandins as signals between tumor and the host immune system.

The development of malignant tissue in vivo is partially favored by the immunosuppression that occurs in cancer patients. However, the signals between tumor and immune tissues remain to be identified. We present evidence that prostaglandins may act as one of these signals by a direct action on cells of the immune system, or by inhibition of insulin secretion which in turn suppresses immune function, or both.

The effect of propionate on lipid synthesis in rat lymphocytes.

1. The effect of propionate on lipid synthesis in lymphocytes cultured for 24 hr and incubated for 2 hr was investigated. 2. [1-14C]-propionate was incorporated mainly into phospholipids in both control and concanavalin A (Con A) stimulated cultured lymphocytes. 3. The content of free coenzyme A markedly decreased in 2 hr incubated lymphocytes when propionate was added to the medium at concentrations from 10 to 100 mmol/l. 4. Propionate at 40 mmol/l decreased the incorporation of [1-14C]-palmitate into phospholipids (86%), triacylglycerol (87%) and cholesterol ester (98%) and increased in cholesterol (133%) of cultured lymphocytes. 5. Addition of propionate into the culture medium at 2.5 and 5.0 mmol/l concentrations markedly increased the activity of hydrolases of various acylCoA derivatives. 6. The results suggest that propionate may reduce the content of acylCoA and so its esterification and this might be important for the regulation of lymphocytes proliferation.

Pyruvate is a lipid precursor for rat lymphocytes in culture: evidence for a lipid exporting capacity.

Since acetyl-CoA produced through pyruvate dehydrogenase reaction is poorly oxidized by the Krebs cycle in rat lymphocytes, the fate of acetyl units was investigated in these cells. The results presented here show that 24-h cultured lymphocytes actively synthesize lipids from [3-14C]pyruvate. Furthermore, a considerable amount of these lipids have shown to be exported into the culture medium. Experiments with [1-14C] acetate as a lipid precursor showed a close similarity with the rates of incorporation of [3-14C] pyruvate into the same lipid fractions. Treatment of lymphocytes with the mitogen concanavalin A (Con A) markedly enhanced [1-14C] acetate incorporation into a variety of lipids, but the lectin did not affect [3-14C] pyruvate incorporation. The results suggest that lymphocytes convert pyruvate into lipids via the acetyl-CoA pathway and that Con A interferes in lymphocyte lipogenesis but does not seem to affect the pyruvate dehydrogenase reaction. The ability to incorporate pyruvate into certain lipids may have an important role for the rapidly dividing capacity of lymphocytes since the human cancer strain HeLa 155 (a quickly proliferating cell line) also exhibits this feature by converting much more [3-14C] pyruvate into lipids than do lymphocytes. In addition, comparative experiments with lymphocytes, peritoneal macrophages and HeLa cells indicate that pyruvate may provide precursors for cells with active lipid producing and exporting capacities.