Haem Biosynthesis and Antioxidant Enzymes in Circulating Cells of Acute Intermittent Porphyria Patients.

The aims of the present study were to explore the expression pattern of haem biosynthesis enzymes in circulating cells of patients affected by two types of porphyria (acute intermittent, AIP, and variegate porphyria, VP), together with the antioxidant enzyme pattern in AIP in order to identify a possible situation of oxidative stress. Sixteen and twelve patients affected by AIP and VP, respectively, were analysed with the same numbers of healthy matched controls. Erythrocytes, neutrophils and peripheral blood mononuclear cells (PBMCs) were purified from blood, and RNA and proteins were extracted for quantitative real time PCR (qRT-PCR) and Western-blot analysis, respectively. Porhobilinogen deaminase (PBGD) and protoporphyrinogen oxidase (PPOX) gene and protein expression was analysed. Antioxidant enzyme activity and gene expression were additionally determined in blood cells, together with protein carbonyl content in plasma. PBMCs isolated from AIP patients presented low mRNA levels of PBGD when compared to controls, while PBMCs isolated from VP patients presented a decrease in PPOX mRNA. PPOX protein content was higher in AIP patients and lower in VP patients, compared to healthy controls. Regarding antioxidant enzymes, PBMCs and erythrocyte superoxide dismutase (SOD) presented statistically significant higher activity in AIP patients compared to controls, while catalase activity tended to be lower in these patients. No differences were observed regarding antioxidant gene expression in white blood cells. Circulating cells in AIP and VP patients present altered expression of haem biosynthetic enzymes, which could be useful for the differential diagnosis of these two types of porphyria in certain difficult cases. AIP patients present a condition of potential oxidative stress similar to VP patients, evidenced by the post-transcriptional activation of SOD and possible catalase impairment.

Exercise in a hot environment influences plasma anti-inflammatory and antioxidant status in well-trained athletes.

Exercise in thermally stressful environmental conditions can enhance oxidative stress. We sought to measure the plasma antioxidant defenses and cytokine response together with oxidative damage post-exercise in a temperate versus a hot environment. The plasma concentrations of vasoactive endothelin-1 and vascular angiogenic growth factor were also evaluated. Male athletes (n=9) volunteered to participate. The athletes randomly performed two bouts of treadmill exercise of 45min at 75-80% of maximal oxygen uptake in a climatic-controlled chamber under two different conditions: temperate environment (10-12°C, 40-55% humidity) and hot, humid environment (30-32°C, 75-78% humidity). Venous blood samples were obtained immediately pre- and post-bout and on recovery after 2h. Serum glucose, malondialdehyde and lactate concentrations were significantly increased post-exercise in hot but maintained in the temperate environment; these post-exercise values were significantly higher after exercise in hot than in temperate. Urinary 8-hydroxy-2'-deoxyguanosine concentration, plasma phosphocreatine kinase and catalase activities, creatinine and monocyte chemoattractant protein-1, and interleukin-6 significantly increased post-exercise in hot but maintained in temperate environment. The post-exercise circulating values of antioxidant enzyme paraoxonase-1 and endothelin were significantly higher in the hot than in temperate environment. Exercise in a hot and humid environment resulted in mild hyperthermia with elevated perceived exertion and thermal stress. Hyperthermic environment induced hyperglycemia, lactatecidemia and more cellular and oxidative damage than exercise in a temperate environment but also induced a post-exercise antioxidant and anti-inflammatory response in plasma. These results suggest that environmental temperature needs to be taken into account when evaluating exercise-related oxidative stress and inflammation.

Body temperature modulates the antioxidant and acute immune responses to exercise.

The aim of this study was to determine the effects of whole body heat in combination with exercise on the oxidative stress and acute phase immune response. Nine male endurance-trained athletes voluntarily performed two running bouts of 45 minutes at 75-80% of VO(2max) in a climatic chamber in two conditions: cold and hot humid environment. Leukocyte, neutrophil and basophil counts significantly rose after exercise in both environments; it was significantly greater in the hot environment. Lymphocyte and neutrophil antioxidant enzyme activities and carbonyl index significantly increased or decreased after exercise only in the hot environment, respectively. The lymphocytes expression of catalase, Hsp72 and CuZn-superoxide dismutase was increased in the hot environment and Sirt3 in the cold environment, mainly during recovery. In conclusion, the increased core body temperature results in the acute phase immune response associated to intense exercise and in the immune cell adaptations to counteract the oxidative stress situation.

Phytoestrogens enhance antioxidant enzymes after swimming exercise and modulate sex hormone plasma levels in female swimmers.

Our aim was to investigate the effects of diet supplementation with phytoestrogens on sex hormone levels, antioxidant adaptive responses and oxidative damage induced by exercise. Ten female swimmers participated for 26 days in a diet intervention with either a functional beverage rich in vitamins C and E or the same beverage but also supplemented with Lippia citriodora extract (PLX) containing 20 mg/100 ml verbascoside. After the intervention all subjects participated in a swimming session for 30 min maintaining the intensity at about 75-80% of their individual best performance time for a 50-m swim. In lymphocytes, the superoxide dismutase activity increased after exercise, with a higher increase in the PLX group. Swimming increased the erythrocyte activity of glutathione peroxidase and glutathione reductase in the PLX group. Purified glutathione reductase activity increased after an in vitro incubation with PLX. No effects were observed on the lymphocyte levels of malondialdehyde and carbonyls, but exercise increased the percentage of high-damaged lymphocytes 2.8 times in the placebo group and 1.5 times in the PLX group. PLX decreased the levels of 17-ß-estradiol and testosterone and increased the levels of the sex hormone binding globulin. In conclusion, supplementation with phytoestrogens enhances the glutathione-dependent enzyme activities in erythrocytes and the superoxide dismutase activity in lymphocytes in response to exercise. PLX also shows direct antioxidant properties, by increasing glutathione reductase enzyme activity in vitro. Supplementation with phytoestrogens also decreases the plasma steroid hormone levels, pointing towards a possible agonistic effect of verbascoside in the hypothalamic regulation of estradiol synthesis.