Relation of CuZnSOD activity with renal insufficiency in hypertensive diabetic patients.

Diabetes and renal insufficiency are interrelated metabolic disorders closely associated with redox homeostasis disturbances. The aim of this study was to compare the activity of copper zinc superoxide dismutase (CuZnSOD) in the erythrocytes of hypertensive diabetic patients with or without renal insufficiency with normal healthy control subjects. In both groups of diabetic patients, blood glucose level and the content of glycosylated hemoglobin (HbA1c) were higher than in the control group. However, CuZnSOD activity was significantly higher than control only in hypertensive diabetic patients with renal insufficiency. Our results suggest that disturbances in superoxide homeostasis do correlate with long-term complication in diabetes, i.e. diabetic renal insufficiency and hypertension.

Detection, amplification & sequence homology of sodC in clinical isolates of Salmonella sp.

Background & objectives: Periplasmic copper and zinc superoxide dismutase (Cu,Zn-SOD or SodC) is an important component of the antioxidant shield which protects bacteria from the phagocytic oxidative burst. Cu,Zn-SODs protect Gram-negative bacteria against oxygen damage which have also been shown to contribute to the pathogenicity of these bacterial species. We report the presence of SodC in drug resistant Salmonella sp. isolated from patients suffering from enteric fever. Further sodC was amplified, cloned into Escherichia coli and the nucleotide sequence and amino acid sequence homology were compared with the standard strain sSalmonella Typhimurium 14028. Methods: Salmonella enterica serovar Typhi (S. Typhi) and Salmonella enterica serovar Paratyphi (S. Paratyphi) were isolated and identified from blood samples of the patients. The isolates were screened for the presence of Cu, Zn-SOD by PAGE using KCN as inhibitor of Cu,Zn-SOD. The gene (sodC) was amplified by PCR, cloned and sequenced. The nucleotide and amino acid sequences of sodC were compared using CLUSTAL X. Results: SodC was detected in 35 per cent of the Salmonella isolates. Amplification of the genomic DNA of S. Typhi and S. Paratyphi with sodC specific primers resulted in 519 and 515 bp amplicons respectively. Single mutational difference at position 489 was observed between the sodC of S. Typhi and S. Paratyphi while they differed at 6 positions with the sodC of S. Typhimurium 14028. The SodC amino acid sequences of the two isolates were homologous but 3 amino acid difference was observed with that of standard strain S. Typhimurium 14028. Interpretation & conclusions: The presence of SodC in pathogenic bacteria could be a novel candidate as phylogenetic marker.

Study Progress of Cu,Zn Superoxide Dismutas——From Gene to Function / 现代生物医学进展

Superoxide Dismutase (SOD)(EC 1.15.1.1)is a metalloenzyme that is found in almost all organisms and catalyzes the dismutation of superoxide anion radical to hydrogen peroxide and molecular oxygen. Three unique and highly compartmentalized mammalian SOD have been biochemically and molecularly characterized to date: Cu, Zn superoxide dismutase (CuZnSOD, SOD1), MnSOD (Manganese Superoxide Dismutase, SOD2)and EC-SOD (Extracellular Superoxide Dismutase, SOD3). Cu, Zn superoxide dismutase (CuZnSOD, SOD1)is a copper and zinc-containing homodimer that is found almost exclusively in intracellular cytoplasmic spaces. CuZnSOD is widely distributed and comprises about 90% of the total SOD. Cytoplasmic and periplasmic SOD exists as dimers,whereas chloroplastic and extracellular enzymes exist as tetramers. Structure supports independent functional evolution in prokaryotes and eukaryotes. CuZnSOD are thought to protect the brain, lungs, and other tissues from oxidative stress. This paper reviewed the gene, molecular and chemical structure and biological function of CuZnSOD.

Study Progress of Cu,Zn Superoxide Dismutas——From Gene to Function / 现代生物医学进展

Superoxide Dismutase (SOD)(EC 1.15.1.1)is a metalloenzyme that is found in almost all organisms and catalyzes the dismutation of superoxide anion radical to hydrogen peroxide and molecular oxygen. Three unique and highly compartmentalized mammalian SOD have been biochemically and molecularly characterized to date: Cu, Zn superoxide dismutase (CuZnSOD, SOD1), MnSOD (Manganese Superoxide Dismutase, SOD2)and EC-SOD (Extracellular Superoxide Dismutase, SOD3). Cu, Zn superoxide dismutase (CuZnSOD, SOD1)is a copper and zinc-containing homodimer that is found almost exclusively in intracellular cytoplasmic spaces. CuZnSOD is widely distributed and comprises about 90% of the total SOD. Cytoplasmic and periplasmic SOD exists as dimers,whereas chloroplastic and extracellular enzymes exist as tetramers. Structure supports independent functional evolution in prokaryotes and eukaryotes. CuZnSOD are thought to protect the brain, lungs, and other tissues from oxidative stress. This paper reviewed the gene, molecular and chemical structure and biological function of CuZnSOD.

Gene Transfer of Cu/ZnSOD to Cerebral Vessels Prevents Subarachnoid Hemorrhage-induced Cerebral Vasospasm

The preventive effects of gene transfer of human copper/zinc superoxide dismutase (Cu/ZnSOD) on the development of cerebral vasospasm after subarachnoid hemorrhage (SAH) were examined using a rat model of SAH. An experimental SAH was produced by injecting autologous arterial blood twice into the cisterna magna, and the changes in the diameter of the middle cerebral artery (MCA) were measured. Rats subjected to SAH exhibited a decreased diameter with an increased wall thickness of MCA that were significantly ameliorated by pretreatment with diphenyleneiodonium (DPI, 10microM), an inhibitor of NAD (P) H oxidase. Furthermore, application of recombinant adenovirus (100microliter of 1 x 1010 pfu/ml, intracisternally), which encodes human Cu/ZnSOD, 3 days before SAH prevented the development of SAH-induced vasospasm. Our findings demonstrate that SAH-induced cerebral vasospasm is closely related with NAD (P) H oxidase-derived reactive oxygen species, and these alterations can be prevented by the recombinant adenovirus-mediated transfer of human Cu/ZnSOD gene to the cerebral vasculature.

The Role of Extracellular Superoxide Dismutase (EC-SOD) in Skeletal Muscle Ischemia-reperfusion Injury / 대한정형외과학회잡지

PURPOSE: This study investigated the role of extracellular superoxide dismutase (EC-SOD), which is a major extracellular antioxidant enzyme in skeletal muscle ischemia and reperfusion (I/R) injury. MATERIALS AND METHODS: The pedicled cremaster muscle flaps from homozygous EC-SOD knockout (EC-SOD-/-), heterozygous CuZn-SOD knockout (CuZn-SOD+/-) and wild-type (WT) mice were subjected to 4.5 hour ischemia followed by 90 min reperfusion. The pedicled cremaster muscle flaps were examined by functional analysis during the reperfusion. The mRNA and protein expressions of each SOD after I/R were evaluated using quantitative real-time PCR and western blot. RESULTS: The results showed that the EC-SOD-/- mice had a more profound I/R injury than the CuZn- SOD+/- or WT mice. In particular, there was a delayed and incomplete recovery of the arterial diameter and blood flow during reperfusion and as well as there being more severe inflammation. After 90 min reperfusion, the EC-SOD mRNA levels increased more in the CuZn-SOD+/- mice than in the WT mice. However, the CuZn-SOD and Mn-SOD mRNA levels decreased similarly in all 3 groups. The CuZn-SOD protein levels decreased in all groups. The EC-SOD protein levels decreased in the CuZn-SOD+/- and WT mice, but the Mn-SOD protein levels were unchanged or slightly increased in all groups. The histological results showed diffuse edema and inflammatory cell infiltration around the muscle fibers and these changes were more severe in the EC-SOD-/- mice. CONCLUSION: EC-SOD plays an important role in protecting the skeletal muscle from I/R injury caused by the excessive generation of reactive oxygen species.

Effects of the pretreatment of free radical scavengers on the expression of apoptosis after gamma-irradiation in the rat parotid gland / 대한해부학회지

Apoptosis is a distinct morphological form of programmed cell death during growth, development and some diseases. Also, it is well known that the apoptosis is usually caused by carcinogens, drugs, heat and radiation in mature cells. Especially, the therapeutic radiation of the head and neck region may cause xerostomia after irradiation. Recently, there are many trial reports to explain the radiation cell death to the acute cellular changes as a programmed cell death. The author performed present study to investigate the potential role of reactive oxygen metabolites as a signal for radiation-induced apoptosis. A series of free radical scavengers, allopurinol, superoxide dismutase, dimethylthiourea were pretreated before gamma-irradiation for evaluating their ability to block apoptosis in the rat parotid gland in vivo, and the light microscopic results were evaluated and analyzed using the Tdt-mediated dUTP-biotin nick end labeling technique. The results were as follows. 1. Under the light microscope, irradiation induced apoptotic bodies in the parotid gland were revealed the intensively positive reaction on the TUNEL technique, and most apoptotic bodies were confined in the acinar system, there were rare apoptotic changes in the ductal system. 2. In all irradiated experimental groups without antioxidants pretreatment, the number of the apototic bodies was the greatest in post-irradiation 24 hour group, and then decreased as a function of time. 3. As a result on pretreatment of allopurinol that effectively inhibit xanthine oxidase before irradiation, failed to block the irradiation-induced apoptosis in the rat parotid gland. Moreover, in the postirradiation 1 day group, the apoptotic expression and destruction of the acini increased than non-irradiated experimental group. 4. As a result on pretreatment of Cu/Zn-SOD that contributes to extracellularly generated superoxides and dimethylthiourea to scavenge the hydroxyl free radicals before irradiation, two antioxidants inhibit the irradiation-induced apoptosis (P<0.001), salivary acinar structures remained intact. 5. To evaluate the synergistic effects, simultaneous pretreatments of allopurinol and Cu/Zn-SOD, Cu/Zn-SOD and DMTU were conducted. Expression of irradiation-induced apoptosis was lower than the experimental groups which not pretreated any antioxidants (P<0.001), but there was no synergism of two antioxidants. 6. Irradiation-induced apoptotic bodies crowded at the outer cortical layer including lymphatic nodules, and the aspect of apoptotic expression in the lymph node took place earlier than in the salivary acini. Taken all together, it is forecasted that the irradiation-induced apoptosis of the parotid gland is a result of primary cellular damage by oxygen or hydroxyl free radicals. Through this study, the facts that pretreatments of free radical scavengers block the irradiation-induced apoptosis were identified. But, oxygen free radicals were not produced via xanthine oxidase process, whereas, allopurinol that inhibits this reaction influenced the harmful effects on the parotid gland.

Superoxide Dismutase Gene Expression Induced by Lipopolysaccharide in Alveolar Macrophage of Rat / 결핵

BACKGROUND: In the pathogenesis of acute lung injury induced by lipopolysaccharide(LPS), oxygen radiclls are known to be involved in one part. Superoxide dismutase(SOD) protects oxygen radical-induced tissue damage by dismutating superoxide to hydrogen peroxide. In eukaryotic cells, two forms of SOD exist intracellularly as a cytosolic, dimeric copper/zinc-containing SOD (CuZnSOD) and a mitochondrial, tetrameric manganese-containing SOD(MnSOD). But there has been little information about SOD gene expression and its regulation in pulmonary alveolar macrophages(PAMs). The objective of this study is to evaluate the SOD gene expression induced by LPS and its regulation in PAMs of rat. METHOD: In Sprague-Dawley rats, PAMs obtained by broncholaveolar lavage were purified by adherence to plastic plate. To study the effect of LPS on the SOD gene expression of PAMs, they were stimulated with different doses of LPS(0.01microg/ml ~ 10microg/ml) and for different intervals(0, 2, 4, 8, 24hrs). Also for evaluating the level of SOD gene regulation actinomycin D(AD) or cyclo- heximide(CHX) were added respectively. To assess whether LPS altered SOD mRNA stability, the rate of mRNA decay was determined in control group and LPS-treated group. Total cellular RNA extraction by guanidinium thiocyanate/phenol/chloroform method and Northern blot analysis by using a 32P-labelled rat MnSOD and CuZnSOD cDNAs were performed. RESULTS: The expression of mRNA in MnSOD increased dose-dependently, but not in CuZnSOD. MnSOD mRNA expression peaked at 8 hours after LPS treatment. Upregulation of MnSOD mRNA expression induced by LPS was suppressed by adding AD or CHX respectively. MnSOD mRNA stability was not altered by LPS. CONCLUSION: These findings show that PAMs of rat could be an important source of SOD in response to LPS, and suggest that their MnSOD mRNA expression may be regulated transcriptionally and require de novo protein synthesis without affecting mRNA stability.