• Toxicological Research
• /
• 제34권4호
• /
• pp.363-370
• /
• 2018

Many studies reported reduced antioxidant capacity in the vasculature under hypertensive conditions. However, little is known about the effects of antihypertensive treatments on the regulation of vascular antioxidant enzymes. Thus, we hypothesized that antihypertensive treatments prevent the reduction of antioxidant enzyme activity and expression in the small vessels of angiotensin II-induced hypertensive rats (ANG). We observed the small mesenteric arteries and small renal vessels of normotensive rats (NORM), ANG, and ANG treated with a triple antihypertensive therapy of reserpine, hydrochlorothiazide, and hydralazine (ANG + TTx). Systolic blood pressure was increased in ANG, which was attenuated by 2 weeks of triple therapy (127, 191, and 143 mmHg for NORM, ANG, and ANG + TTx, respectively; p < 0.05). Total superoxide dismutase (SOD) activity in the small mesenteric arteries of ANG was lower than that of NORM. The protein expression of SOD1 was lower in ANG than in NORM, whereas SOD2 and SOD3 expression was not different between the groups. Reduced SOD activity and SOD1 expression in ANG was not restored in ANG + TTx. Both SOD activity and SOD isoform expression in the small renal vessels of ANG were not different from those of NORM. Interestingly, SOD activity in the small renal vessels was reduced by TTx. Between groups, there was no difference in catalase activity or expression in both the small mesenteric arteries and small renal vessels. In conclusion, SOD activity in the small mesenteric arteries decreased by angiotensin II administration, but not by hypertension, which is caused by decreased SOD1 expression.

• BMB Reports
• /
• 제46권11호
• /
• pp.555-560
• /
• 2013

Acrolein is the most reactive aldehydic product of lipid peroxidation and is found to be elevated in the brain when oxidative stress is high. The effects of acrolein on the structure and function of human Cu,Zn-superoxide dismutase (SOD) were examined. When Cu,Zn-SOD was incubated with acrolein, the covalent crosslinking of the protein was increased, and the loss of enzymatic activity was increased in a dose-dependent manner. Reactive oxygen species (ROS) scavengers and copper chelators inhibited the acrolein-mediated Cu,Zn-SOD modification and the formation of carbonyl compound. The present study shows that ROS may play a critical role in acrolein-induced Cu,Zn-SOD modification and inactivation. When Cu,Zn-SOD that has been exposed to acrolein was subsequently analyzed by amino acid analysis, serine, histidine, arginine, threonine and lysine residues were particularly sensitive. It is suggested that the modification and inactivation of Cu,Zn-SOD by acrolein could be produced by more oxidative cell environments.

• Journal of Microbiology and Biotechnology
• /
• 제3권3호
• /
• pp.188-193
• /
• 1993

Superoxide dismutase (SOD) was purified from Pseudomonas polycolor to an electrophoretically homogeneous state and partially characterized. SOD was purified by ammonium sulfate fractionation, column chromatography on DEAE-Sephadex A-50, phenyl-Toyopearl 650 M, and gel filtration on Sephadex G-100. The molecular weight and subunit molecular weight of the purified enzyme were estimated to be 40, 000 and 20, 000, respectively. The purified enzyme remained stable at pH 9.0~11.0, $25^{\circ}C$ for 40 hr, but rapidly became inactive below 9.0. SOD was stable up to $45^{\circ}C$ at pH 9.0 with about 80% relative activity, but rapidly became inactive at temperature above that. The enzyme was insensitive to cyanide and fluoride, and sensitive to hydrogen peroxide and azide. The results suggest that the enzyme be an iron-containing SOD.

• 약학회지
• /
• 제48권6호
• /
• pp.309-316
• /
• 2004

The present investigation was undertaken to find out the anticancer activity of monoterpene compounds. Monoterpenes showed generally the inhibitory effect on the proliferation o f L1210 cancer cells (cytotoxicity). Geraniol was found to exibit the most potent cytotoxic effect on L1210 cells with an IC50 values of $0.67{\mu}g/ml$. On the other hand, geraniol proved to be capable of stimulating the macrophage proliferation (135% of control). When the life prolonging activity of geraniol by daily oral administration of 0.1~10${\mu}g/10{\mu}l/20$ g body weight to Sarcoma 180 bearing ICR mouse was examined, there was also a significant elevation of survival (best result of 134% of control). The contradictory effects of geraniol on the proliferation of L1210 cells and macrophages proved to be accompanied by the coincident alterations of RNS (reactive nitrogen species) related enzymes activities such as inducible nitric oxide synthase (Inos) in macrophages and ROS (reactive oxygen species) related enzymes activities such as superoxide dismutase (SOD) in L1210 cells, respectively.

• Journal of Plant Biotechnology
• /
• 제29권1호
• /
• pp.7-13
• /
• 2002

Superoxide dismutase (SOD)를 과실에서 고발현시킨 형질전환 토마토 (서광과 꼬꼬)를 개발하였다. 카사바 배양세포에서 분리한 CuZnSOD (mSOD1)를 과실에 우세적으로 발현하는 ascorbate oxidase promoter (ASOp)를 이용하여 ASOp :: mSOD1/pBI101 벡터를 제작한 후 Agrobacterium 매개로 자엽 절편체를 형질전환하였다. Kanamycin 저항성 식물체를 기관발생 경로로 재분화시킨 후 Southern 분석으로 형질전환을 확인하였다. 서광과 꼬꼬 토마토의 형질전환체와 대조구 식물체의 과실을 성숙 단계별로 분류하여 단백질 함량과 SOD 비활성도 (units/mg protein)를 측정한 결과, 단백질 함량은 열매가 익은 단계로 갈수록 점점 감소하여 완전히 익은 단계에서 가장 낮았다. SOD 비활성도는 형질전환 토마토의 열매의 모든 단계에서 대조구보다 높았으며 완전히 성숙한 과실에서 가장 높았다. 성숙한 형질전환 서광과 꼬꼬 과실에서 SOD 비활성도는 비형질전환의 것보다 각각 약 1.6배와 약 2.2배 높았다. SOD isoenzyme gel 분석에서 도입한 mSOD1로 추정되는 CuZnSOD 밴드가 형질전환체에서 과실 성숙에 따라 강하게 발현되었다. 이상의 결과로서 ASO promoter에 의해 SOD 유전자가 토마토 과실에 특이적으로 발현됨이 확인되었다.

• Bulletin of the Korean Chemical Society
• /
• 제22권4호
• /
• pp.362-366
• /
• 2001

To show the effects of metallothionein (MT) on the activity of enzymes involved in the removal of reactive oxygen species, MT has been added to the assay systems of superoxide dismutase (SOD), catalase and peroxidase. We have used assay systems of SOD based on NADPH oxidation and nitrite formation from hydroxylammonium chloride as an assay of superoxide breakdown rate. The two assay systems showed different results at the high concentration of MT. MT showed the scavenging of superoxide in the SOD assay system in the presence and absence of SOD. MT added to the SOD assay system behaved as an activator of SOD, but apo-MT behaved as an inhibitor. When MT was added to the assay system in the presence of a fixed amount of SOD, the breakdown rate of superoxide increased. The effects of MT on the decomposition of hydrogen peroxide and the activity of catalase and peroxidase decomposing hydrogen peroxide were evaluated. MT decreased the activities of catalase and peroxidase. We have concluded that the function of MT as an antioxidant might effect the level of superoxide scavenging and not the level of hydrogen peroxide.

• 대한약리학회지
• /
• 제32권2호
• /
• pp.259-267
• /
• 1996

In an attempt to dofine the early biochemical determinants that participate in the pathogenesis of glycerol-induced nephrotoxicity, especially focusing on oxygen free radicals and $N-acetyl-{\beta}-D-glucosaminidase$ (NAG) activity, we studied 24-hours urine outflow, 24-hours urinary protein excretion and urinary NAG activity after the injection of glycerol and also we studied malondialdehyde(MDA) level and superoxide dismutase(SOD) activity in the kidney of rats at 24hr after the injection of glycerol. Sprague-Dawley albino rats weighing 240 to 260 gm were injected intramuscularly with a 50% solution of glycerol(2ml/kg, 4ml/kg and 8ml/kg). The group treated with glycerol showed significantly lower urine outflow level and urinary protein excretion level and higher urinary NAG activity after the injection as compared to those of control group. Also the group treated with glycerol showed significantly higher MDA level and lower SOD activity at 24hr after the injection as compared to those of control group. These results suggest that the excessive oxygen free radicals resulting from the depression of SOD activity is an important determinant in the pathogenesis of glycerol-induced nephrotoxicity and higher urinary NAG activity is an index of renal tubular cell damage in the glycerol-induced nephrotoxicity.

• BMB Reports
• /
• 제30권1호
• /
• pp.60-65
• /
• 1997

Expression of human Cu.Zn-superoxide dismutase (SOD) with activity comparable to human erythrocyte enzyme was achieved in E. coli B21(DE3) by using the pET-17b expression vector containing a T7 promoter. Recombinant human SOD was found in the cytosol of disrupted bacterial cells and represented > 25% of the total bacterial proteins. The protein produced by the E. coli cells was purified using a combination of ammonium sulfate precipitation, Sephacryl S-100 gel filtration and DEAE-Sephacel ion exchange chromatography. The recombinant Cu,Zn-SOD and human erythrocyte enzyme were compared using dismutation activity, SDS-PAGE and immunoblotting analysis. The mass of the subunits was determined to be 15,809 by using a electrospray mass spectrometer. The copper specific chelator. diethyldithiocarbamate (DOC) reacted with the recombinant Cu,Zn-SOD. At $50{\mu}M$ and $100{\mu}M$ concentrations of DOC, the dismutation activity was not inhibited for one hour but gradually reduced after one hour. This result suggests that the reaction of DOC with the enzyme occurred in two distinct phases (phase I and phase II). During phase I of this reaction, one DOC reacted with the copper center, with retention of the dismutation activity while the second DOC displaced the copper, with a loss of activity in phase II.

• 식물조직배양학회지
• /
• 제24권1호
• /
• pp.57-61
• /
• 1997

Superoxide dismutase (SOD) 고생산세포주로 선발된 토마토(Lycopersicun esculentum) 배양세포를 사용하여 현탁배양에 따른 SOD 활성과 isoenzyme변화를 조사하고 토마토 식물체의 것과 비교하였다. 현탁배양은 세포생중량 2 g을 1 mg/L 2,4-D, 30 g/L sucrose를 함유한 MS 배지 50 mL과 함께 mL flask에서 $25^{\circ}C$암상태로 배양(100 rpm)하였다. 세포생장은 계대배양후 20일에 최고점에 도달한 후, 급격히 감소하며 배양 후 23일부터 세포가 검게 변하였다. 세포 단위무게당 SOD활성(unit/g dry cell wt)은 배양 후 23일부터 증가하여 28일째에 최고활성(52,400 unit)을 나타낸 후 급격히 감소하였다. 세포 밖으로 분비되는 extracellular SOD활성은 배양 후 25일에 최고치(27,800 unit/so mL medium)를 나타낸 후 감소하였다. Flask 전체의 SOD활성은 배양 후 25일에 최대치(35,700 unit)를 나타내었으며 extracellular SOD 활성이 약 75%을 차지하였다. 토마토 배양세포에는 4개의 SOD isoenzyme이 존재하며, isoenzyme의 패턴변화는 세포생장에 따른 효소활성의 변화와 일치하였다. 토마토 식물체는 배양세포에 없는 CuZnSOD가 존재하며 배양세포와 식물체 조직사이에는 서로 다른 isoenzyme 패턴이 존재함을 알 수 있었다.

• Journal of Crop Science and Biotechnology
• /
• 제10권1호
• /
• pp.8-12
• /
• 2007

All accessions of Rehmannia glutinosa show the unique characteristic of intrinsic tolerance to paraquat. The higher level of endogenous superoxide dismutase(SOD) activity and its increase upon paraquat treatment indicated the involvement of SOD in the tolerance mechanism to paraquat in R. glutinosa. In this study, we examined the isoform-specific response of SOD to oxidative stresses and hormones. Six SOD isoforms were found in the leaf, and they were identified as two MnSODs(named MnSOD I and MnSOD II, in order of increasing mobility), one FeSOD and three Cu/ZnSODs(named Cu/ZnSOD I, Cu/ZnSOD II, and Cu/ZnSOD III, in order of increasing mobility). MnSOD I, MnSOD II, FeSOD, Cu/ZnSOD I, Cu/ZnSOD II, and Cu/ZnSOD III, contributed to 4, 11, 7, 15, 30, and 32% of the total SOD activity, respectively. Total SOD activity levels in the leaf were increased by 4, 24, and 21% by paraquat, salicylic acid(SA), and yeast extract(YE), respectively, but little by ethephon. Six SOD isoforms responded differentially to these stresses and hormones. The activities of all the isoforms were increased by YE and SA except that of MnSOD I which was decreased by SA. The activities of MnSOD I, FeSOD, and CuZnSOD I were increased by paraquat. These results suggest that amelioration of oxidative stresses by SOD is fine-tuned by the differential expression of isoforms in R. glutinosa.