Fine-structural distribution of MMP-2 and MMP-9 activities in the rat skeletal muscle upon training: a study by high-resolution in situ zymography

Matrix metalloproteinases (MMPs) are key regulators of extracellular matrix remodeling, but have also important intracellular targets. The purpose of this study was to examine the activity and subcellular localization of the gelatinases MMP-2 and MMP-9 in skeletal muscle of control and physically trained rats. In control hind limb muscle, the activity of the gelatinases was barely detectable. In contrast, after 5 days of intense exercise, in Soleus (Sol), but not Extensor digitorum longus (EDL) muscle, significant upregulation of gelatinolytic activity in myofibers was observed mainly in the nuclei, as assessed by high resolution in situ zymography. The nuclei of quiescent satellite cells did not contain the activity. Within the myonuclei, the gelatinolytic activity colocalized with an activated RNA Polymerase II. Also in Sol, but not in EDL, there were few foci of mononuclear cells with strongly positive cytoplasm, associated with apparent necrotic myofibers. These cells were identified as activated satellite cells/myoblasts. No extracellular gelatinase activity was observed. Gel zymography combined with subcellular fractionation revealed training-related upregulation of active MMP-2 in the nuclear fraction, and increase of active MMP-9 in the cytoplasmic fraction of Sol. Using RT-PCR, selective increase in MMP-9 mRNA was observed. We conclude that training activates nuclear MMP-2, and increases expression and activity of cytoplasmic MMP-9 in Sol, but not in EDL. Our results suggest that the gelatinases are involved in muscle adaptation to training, and that MMP-2 may play a novel role in myonuclear functions

Effect of D-penicillamine on rat lung elastin cross-linking during the perinatal period

This study was designed to clarify the effects of D-penicillamine (DPA), a drug used for treatment of various pathological events, on lung elastin formation and maturation of the newborn in the perinatal period The investigation was conducted on 20 newborn rats bred from 40 female and six male rats. DPA doses 400 mg kg(-1) day(-1) and physiological saline were given intraperitoneally (i.p) to experimental and control goups. To assess newborn maturation, their body and lung weights were determined. Serum Cu levels were measured by atomic absorption spectroscopy and ceruloplasmin (Cp) activities were measured spectrophotometrically. Newborn lung tissue elastin, desmosine (DES) and isodesmosine (IDES) levels were measured by HPLC. The results showed that DPA treatment caused loss of skin elasticity and reduction in body and lung weight in newborns of the experimental group. The serum Cu levels and Cp activity were found to be significantly lower in both maternal and newborn of the experimental groups compared with the control group. The lung DES, IDES and elastin values of newborns in the experimental group were decreased compared with the control group. In conclusion, our results indicate that 400mg kg(-1) day(-1) DPA, a dose that is used in the treatment of Wilson's disease, rheumatoid arthritis and cystinuria, caused the retardation of newborn maturation, a decrease in DES-IDES cross-links and levels of lung elastin of offspring in the perinatal period. Another conclusion to be drawn from this study is that even low levels of Cu depletion due to DPA administration induces a change in cross-linking in lung elastin during the perinatal period. Copyright (c) 2005 John Wiley & Sons, Ltd

Dietary vitamin E (alpha-tocopheryl acetate) and organic selenium supplementation: performance and antioxidant status of broilers fed n-3 PUFA-enriched feeds

WOS: 000275211300004The experiment was conducted to study the effects of organic selenium (Se-enriched yeast) and vitamin E (alpha-tocopheryl acetate) supplementation, alone or together, on the performance and antioxidant status of broilers fed diets enriched in n-3 PUFAs using fish oil. Day-old Hubbard-JV strain broiler chicks allocated to one of the following treatments: (1) a commercial basal diet containing 0.15 mg inorganic Se/kg as sodium selenite and 50 mg alpha-tocopheryl acetate (alpha-TA)/kg feed acted as the control; (2) VE200: Control diet supplemented with 200 mg alpha-TA/kg feed; (3) OrgSe0.15: Control with inorganic Se replaced with 0.15 mg organic Se/kg; (4) OrgSe0.30: Control with inorganic Se replaced with 0.30 mg organic Se/kg feed; (5) VE200+OrgSe0.15, (6) VE200+OrgSe0.30. Body weight (BW) and body weight gain (BWG) to 42 d were significantly improved with vitamin E or organic Se at 0.20 mg/kg (but not at 0.15 mg/kg) when supplemented individually, but not in combination, except that VE200+OrgSe0.15 improved 28 and 35-d body weights significantly compared with the control. Feed intake (FI), feed conversion ratio (FCR), mortality, carcass characteristics and relative organ weights, except for the spleen, were unaffected by any of the supplementation treatments. The significant positive effect of VE200 on BWG was observed after 21 d. Relative spleen weight was significantly higher in broilers fed VE200 compared to the other treatments. The antioxidative effects of organic Se and vitamin E, alone or together, were not evident in diets enriched in n-3 PUFAs using 1.5% fish oil.Scientific Research Projects Committee at Ege UniversityEge University [2003-ZRF-021]This study was funded by the Scientific Research Projects Committee at Ege University (Project number 2003-ZRF-021). We wish to thank Alltech Turkey for supplying Sel-Plex and Eryas firm, Turkey for preparing mineral-vitamin premixes used in the study

Effect of melatonin cotreatment against kainic acid on Coenzyme Q10, lipid peroxidation and Trx mRNA in rat hippocampus

PubMed ID: 15370175Oxidative stress is a likely molecular mechanism in the neurotoxicity of kainic acid (KA), an excitotoxic substance. The aim of this report was to assess the effect of melatonin co-treatment against KA by measuring the levels of Coenzyme Q10 (CoQ 10), lipid peroxidation (LPO), and Thioredoxin (Trx) mRNA in the rat hippocampus. The male rats were divided into three groups as saline, KA treatment (15 mg/kg), and KA plus melatonin (20 mg/kg). The levels of LPO and CoQ10 were determined by high pressure liquid chromatography (HPLC) consisting of fluorescence and electro-chemical detectors, respectively. The expression of the Trx gene was quantified using reverse transcription followed by real-time polymerase chain reaction (RT-PCR). The results show that the level of LPO increased although the level of CoQ10 decreased both in homogenates and mitochondria in KA-treated rats However, melatonin co-treatment attenuated the level of LPO and partially restored the level of CoQ10. Melatonin co-treatment against KA did not affect the regulation of Trx. Finally, in the context of the decreased LPO and the increased CoQ10, the results suggest that melatonin may be protective against central nervous system pathologies involving excitotoxicity or where oxidative damage may contribute to mitochondrial dysfunction.Received 27 February 2004. This study was supported by Ege University Research Foundation. Address correspondence to Ayfer Yalcin, Pharm., PhD, Assoc. Prof., Department of Biochemistry, Faculty of Pharmacy, Ege University, 35100 Bornova, Izmir, Turkey. E-mail: [email protected] -

Effect of melatonin cotreatment against kainic acid on coenzyme Q10 lipid peroxidation and Trx mRNA in rat hippocampus

Oxidative stress is a likely molecular mechanism in the neurotoxicity of kainic acid (KA), an excitotoxic substance. The aim of this report was to assess the effect qf melatonin co-treatment against KA by measuring the levels of Coenzyme Q10 (CoQ 10), lipid peroxidation (LPO), and Thioredoxin (Trx) mRNA in the rat hippocampus. The male rats were divided into three groups as saline, KA treatment (15 mg/kg), and KA plus melatonin (20 mg/kg). The levels of LPO and CoQ10 were determined by high pressure liquid chromatography (HPLC) consisting of fluorescence and electro-chemical detectors, respectively. The expression of the Trx gene was quantified using reverse transcription followed by real-time polymerase chain reaction (RT-PCR). The results show that the level of LPO increased although the level of CoQ 10 decreased both in homogenates and mitochondria in KA-treated rats However, melatonin co-treatment attenuated the level of LPO and partially restored the level of CoQ10. Melatonin co-treatment against KA did not affect the regulation of Trx. Finally, in the context of the decreased LPO and the increased CoQ10, the results suggest that melatonin may be protective against central nervous system pathologies involving excitotoxicity or where oxidative damage may contribute to mitochondrial dysfunction

Synergistic Effects of Methotrexate and Suberoylanilide Hydroxamic Acid in Triggering Apoptosis of Chronic Myeloid Leukemia Cells

In this study, we have investigated the effects of suberoylanilide hydroxamic acid (SAHA) against chronic myeloid leukemia (CML) cells in combination studies with methotrexate (MTX), which is a dihydrofolate reductase inhibitor used in combination therapy with other agents or alone. Combination of synergistic ratios of MTX and SAHA led to apoptotic cell death of CML cells via PARR cleavage, cytochrome c release and ROS increase in vitro. We suggest that combination of MD( and SAHA may minimize the toxicity and side effects of SAHA treatment, thus providing lower amounts of each drug in CML treatment