Effect of miR-124a on oxidative stress injury of pancreatic tissue in type 2 diabetic mice / 中国药理学通报

Aim To investigate the effect of miR-124a on oxidative stress injury and β-cell function of pancreas in type 2 diabetic mice. Methods The wild-type C57BL/6 mice and the C57BIV6 mice with low expression of miR-124a were randomly divided into two groups, namely wild-type control (WT Con), miR-124a

Effect and mechanism of estrogen on EPCs function in diabetic rats / 药学实践杂志

Objective To explore the effect and mechanism of estrogen on endothelial progenitor cells（EPCs）function in diabetic rats. Methods EPCs were isolated from bone marrow of rats and characterized by fluorescence microscopy and flow cytometry. Rat diabetic model was established via streptozotocin induction. The bone marrow was taken to culture EPCs. EPCs of diabetes were incubated with Estrogen 10 nmol/L for 24h. The functions and proliferation of EPCs in vitro were detected. The levels of MnSOD and NO in EPCs and TSP-1 in supernatant were assayed. Results Compared with control group, EPCs proliferation, adhesion and angiogenesis functions were impaired in diabetic rats. The level of MnSOD and NO in diabetic EPCs were significantly decreased, while TSP-1 protein level in the supernatant increased. The above changes can be reversed with estrogen incubation. Conclusion Estrogen improved the EPCs migration and tubule formation in diabetic rats. The mechanism may be related to the reduction of oxidative stress and downregulation of TSP-1 expression in diabetic EPCs.

The protective effect of N-acetylcysteine on mitochondrial oxidative damage of airway epithelial cells induced by cigarette smoke extract / 西安交通大学学报(医学版)

【Objective】 To investigate the effect of N-acetylcysteine （NAC） on mitochondrial damage of airway epithelial cells induced by cigarette smoke extract （CSE）. 【Methods】 Human airway epithelial cells （BEAS-2B） were cultured and divided into three groups as follows： normal control group， 7.5% （75 mL/L） CSE-treated group and 7.5% CSE plus NAC group. After stimulation for 24 hours， cell viability was determined by MTT， and the levels of mitochondrial reactive oxygen species （ROS） and mitochondrial membrane potential （MMP） were observed under the fluorescence microscope. MMP was also measured by flow cytometry， the protein expressions of Sirt3 and manganese superoxide dismutase （MnSOD） were detected by Western blotting， and MnSOD activity was measured by colorimetry. 【Results】 Pretreatment with NAC significantly improved the viability of airway epithelial cells （P<0.05）. The results of fluorescence microscopy and flow cytometry showed that NAC pretreatment significantly attenuated MMP decline in airway epithelial cells exposed to 7.5% CSE （P<0.05）. Compared with 7.5% CSE-treated group， mitochondrial ROS in airway epithelial cells was significantly decreased in 7.5% CSE plus NAC group （P<0.05）. In addition， pretreatment with NAC significantly inhibited the decrease of Sirt3 and MnSOD protein expression and improved MnSOD activity in airway epithelial cells exposed to 7.5% CSE （P<0.05）. 【Conclusion】 NAC attenuates CSE-induced airway epithelial mitochondrial damage through the regulation of Sirt3-MnSOD signaling pathway， which reveals a new mechanism of NAC treatment for chronic obstructive pulmonary disease.

Effect of Kaixinsan on Keap-1/Nrf2/MnSOD Signaling Pathway in Rats with Alzheimer's Disease Induced by Aβ1-42 / 中国实验方剂学杂志

Objective:To explore the possible mechanism of Kaixinsan in improving cognitive impairment in Alzheimer's disease （AD） model rats based on the epichlorohydrin associated protein-1 （Keap-1）/nuclear factor E2 related factor （Nrf2）/manganese superoxide dismutase （MnSOD） signaling pathway. Method:The AD model was established by injecting Amyloid <italic>β</italic>_1-42 （A<italic>β</italic>_1-42， 5 μL） into the lateral ventricle. After modeling， the experimental rats were randomly divided into model group， donepezil group， and Kaixinsan low dose， medium dose and high dose groups. Another normal control group was also established. The donepezil group received donepezil tablets （1.8 g·kg^-1·d^-1）， Kaixinsan low dose， medium dose and high dose groups received corresponding doses of Kaixinsan （10， 20， 40 g·kg^-1·d^-1， respectively）， and the normal control group and model group were given with equal volume of pure water. Morris water maze was used to test the learning and memory ability of rats. The pathological morphology of hippocampal CA3 area was observed by Nissl staining. The expression levels of myeloperoxidase （MPO）， inducible nitric oxide synthase （iNOS） and superoxide dismutase （SOD） in serum were detected by colorimetry， and the protein expression levels of Keap-1， Nrf2 and MnSOD in hippocampus were detected by immunohistochemistry （IHC） and Western bolt. Result:Compared with the normal control group， the escape latency， total swimming distance and first arrival time of the plateau in the model group increased （<italic>P</italic><0.01）， while the times of crossing the plateau and the time in target quadrant decreased （<italic>P</italic><0.01）. Compared with the model group， the rats in donepezil group and Kaixinsan groups showed less latency， lower total swimming distance and first arrival time on the platform （<italic>P</italic><0.05， <italic>P</italic><0.01）， while the times of crossing the platform and time in target quadrant increased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the normal control group， the expression levels of MPO and iNOS in serum of the model group increased （<italic>P</italic><0.01）， while the expression levels of SOD decreased （<italic>P</italic><0.01）. Compared with model group， the expression of MPO and iNOS in serum of donepezil group and Kaixinsan groups decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）， while the expression of SOD increased （<italic>P</italic><0.05， <italic>P</italic><0.01）. In the normal control group， the neurons in the hippocampal CA3 of the rats were arranged neatly， without obvious Nissl body shrinkage. The neurons in the CA3 of the hippocampus of the model group were not arranged neatly， with obvious neuron loss and pyknosis of Nissl body. The neurons in the CA3 of the hippocampus of the rats in the donepezil group and Kaixinsan groups were arranged neatly， with increased number of neurons and decreased Nissl body shrinkage. Compared with the normal control group， the integrated optical density （<italic>IA</italic>） and protein level of Keap-1 in the hippocampus of the model group decreased（<italic>P</italic><0.01）， while the <italic>IA</italic> and protein level of Nrf2 and MnSOD increased （<italic>P</italic><0.01）. Compared with model group， <italic>IA</italic> and protein levels of Keap-1 and MnSOD in hippocampus of rats in donepezil group and Kaixinsan groups increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， while <italic>IA</italic> and protein levels of Nrf2 decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:Kaixinsan could alleviate memory impairment in AD rats， and its mechanism may be related to its regulation of Keap-1/Nrf2/MnSOD signaling pathway.

Effect of Danggui Buxuetang on PKD1/NF-κB/MnSOD Signal Pathway in Bleomycin-induced Pulmonary Fibrosis in Rats / 中国实验方剂学杂志

Objective:To observe the effect of Danggui Buxuetang on lung histopathology and protein kinase D1 (PKD1), nuclear transcription factor-κB (NF-κB) and manganese superoxide dismutase (MnSOD)-mediated oxidative stress pathway in rats with pulmonary fibrosis induced by bleomycin, so as to explore the mechanism of intervention of pulmonary fibrosis.Method:Thirty-two male SPF SD rats were randomly divided into sham operation group, model group, Danggui Buxuetang group and prednisone group, with 8 rats in each group. Except the sham operation group, the other groups were prepared through the intratracheal instillation with bleomycin. After modeling for 24 h, the rats of Danggui Buxuetang group were administered with Danggui Buxuetang (0.81 g·kg-1). The rats of prednisone group were given aqueous solution of prednisone (0.005 g·kg-1). The rats of sham operation group and model group were given the same volume of saline. After 14 days of administration, blood was collected from the femoral artery, serum was separated, and the lungs were taken by thoracotomy. The pathological changes of rat lung tissues were observed by hematoxylin-eosin staining (HE) and Masson trichrome staining, and graded by Szapiel score and Ashcroft score at the same time. The content of serum malondialdehyde (MDA), and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) were determined. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure mRNA and protein expressions of PKD1, NF-κB, MnSOD.Result:Compared with the rats in sham operation group, the rats in model group had higher Szapiel scores and Ashcroft scores (P<0.05), higher serum MDA content , but lower SOD, CAT and GSH-Px activities(P<0.01), moreover, the rat lung tissues in model group had higher mRNA and protein expressions of PKD1, NF-κB and MnSOD (P<0.01) than those in sham operation group. Compared with the rats in model group, the Szapiel scores and Ashcroft scores of the rats in Danggui Buxuetang group were decreased significantly（P<0.05）. The serum MDA content was decreased significantly, and SOD, CAT, GSH-Px activities were increased, whereas mRNA and protein expressions of PKD1, NF-κB, MnSOD in the rat lung tissues were decreased（P<0.05，P<0.01）.Conclusion:Danggui Buxuetang can reduce the degree of pulmonary fibrosis by regulating the anti-oxidation pathway of PKD1/NF-κB/MnSOD mitochondrial nucleus and improving the body's antioxidant capacity.

The study on the effect of ginkgo biloba extracts on MnSOD activity in the treatment of tardive dyskinesia / 中国神经精神疾病杂志

To investigate if ginkgo biloba extract (Egb-761) can improve tardive dyskinesia (TD) symptoms through increasing the activity of plasma MnSOD. Methods We enrolled a total of 384 schizophrenia patients including 157 TD patients and 227 non-TD patients, as well as 280 normal subjects. The difference of MnSOD level in plasma among these groups were compared. TD patients were then randomly divided into two groups. The treatment group (n=77) and the placebo group (n=75) were treated with 240 mg of Egb-761 or placebo per day for 12 weeks, respectively. The abnormal involuntary movement scale (AIMS) and the positive and negative symptoms scale (PANSS) were used to evaluate the severity of the symptoms in baseline, the sixth week and the twelfth week after treatment. The level of MnSOD activity in plasma was also detected before and after the treatment. Results The level of MnSOD activity was lower in schizophrenia groups than in healthy control group (P＜0.01). In addition, the level of MnSOD activity was significantly lower in TD group than in non-TD group (P＜0.05). Repeated measures analysis of variance showed that group effect (F=4.00, P=0.05), time effect (F=32.17, P＜0.01) and interactive effect of group and time (F=39.04, P＜0.01) were significant in AIMS total score. The AIMS total score of treatment group was significantly lower than that of placebo group at 6-week and 12-week time points (all P＜0.01). Repeated measures analysis of variance showed that time effect (F=23.04, P＜0.01) and interactive effect of group and time (F=6.41, P＜0.05) were significant in the level of MnSOD activity. In addition, the level of MnSOD at baseline was significantly correlated with the reduction of AIMS total score during the treatment period (r=0.27, P=0.018). Conclusion Treatment of Egb-761 can improve symptoms of TD and activity of MnSOD.

Resveratrol mediates apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis by down-regulating mnsod / 中国药理学通报

Aim: To investigate the role of manganese superoxide dismutase (MS0D) protein expression in the proliferation and apoptosis of fibroblast-like synoviocyte (RA-FLS) in rheumatoid arthritis mediated by resveratrol(Res). Methods: After RA-FLSs was treated with Res, the expression of MnSOD protein was detected by CCK-8 method, and the expression of Mn- SOD protein was detected by, Western blot. After lentivirus infection with RA-FLSs, 8 mg · L-1 purine mycin was used to screen the infected cells, and three stable cell lines were established; MnSOD overexpression, MnSOD interference and MnSOD control. The mitochondria reactive oxygen species (mtROS) levels of each cell line treated with 5 μmol · L-1 hydrogen peroxide (H2O2) and 200 mol · L-1 Res were detected by laser confocal microscopy, and cell apoptosis was detected by flow cytometry. Results Res could inhibit the proliferation and the expression of MnSOD in RA- FLSs. The lentivirus-mediated MnSOD regulation could significantly increase or decrease the expression of Mn- SOD in RA-FLSs. MnSOD over expression could decrease the level of mtROS and apoptosis of the RA- FLSs treated with 5 μmol · L-1 H2O2 and 200 μmol · L-1 Res. While the MnSOD RNAi showed the opposite results. Conclusions: Res may up-regulate mtROS levels by inhibiting the expression of MnSOD, thus promoting the apoptosis of RA-FLSs.

Expression, purification, stability and transduction efficiency of full-length SOD2 recombinant proteins / 生物工程学报

Superoxide dismutase (SOD) family is necessary to protect cells from the toxicity of reactive oxygen species produced during normal metabolism. Among SODs, manganese-containing superoxide dismutase (Mn-SOD, SOD2) is the most important one. The DNA fragment containing the full nucleotide of full-length human SOD2 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1 with tag GST. DNA construct was then transformed into Escherichia coli BL21 (DE3) and expression was induced with IPTG at 25 ℃. The recombinant fusion protein GST-SOD2 (46 kDa) was purified from the bacterial lysate by GST resin column affinity chromatography. GST tag was cleaved with thrombin, and a crude SOD2 recombinant protein (25 kDa) was obtained and further purified by heparin affinity chromatography. Activities of the two SOD2 proteins were 1 788 and 2 000 U/mg, respectively. Both SOD2 proteins were stable under physiological condition and cell-penetrating (P<0.05). Our findings open the possibility to study the structure and effects of two full-length recombinant SOD2 proteins.

Study on the Relationship between Lipoprotein Associated Phospholipase A2 Level and Manganese Superoxide Dismutase 9 Ala/Val Genetic Polymorphism in Patients with Coronary Heart Disease / 现代检验医学杂志

Objective To investigate the clinical value of the combined detection of manganese superoxide dismutase (Mn-SOD)genetic polymorphism and lipoprotein phospholipase A2 (Lp-PLA2)in the early diagnosis of coronary heart disease (CHD).Methods 92 cases of coronary heart disease patients and 78 cases of healthy control group were selected.The activ-ity of Lp-PLA2,the activity of Mn-SOD and genotype of Mn-SOD 9 Ala/Val genetic polymorphism were detected in the ser-um of each group via the use of colorimetry,continuous monitoring technique and gene-sequencing method respectively and then the correlation of serum Mn-SOD,Lp-PLA2 and Mn-SOD genetic polymorphism with CHD were analyzed.Results The Lp-PLA2 activity in serum of CHD groups with Mn-SOD 9 VV genotype was statistically significantly higher than that of patients with the AV+AA genotype (P<0.01).The serum Mn-SOD activity in patients with VV genotype was signifi-cantly lower than that of those with AV+AA genotype (P<0.01).Conclusion Combined detection of Mn-SOD 9 Ala/Val genetic polymorphism and Lp-PLA2 activityin the serum can provide an important foundation for the diagnosis and predic-tion of coronary heart disease.

Relationship of Superoxide Dismutase Isoenzyme and High Sensitive C-Reactive Protein with Coronary Heart Disease / 现代检验医学杂志

Objective To investigate the relationship between the activity of superoxide dismutase (T-SOD)as well as its manganese superoxide dismutase isozyme (Mn-SOD),concentration levels of hypersensitive c-reactive protein(hs-CRP)and coronary heart disease.Methods The levels of serum T-SOD,Mn-SOD and hs-CRP were measured in 81 patients with coro-nary heart disease and 60 healthy controls,respectively.T-SOD was measured by colorimetricmethod and hs-CRP was meas-ured by latex enhanced immune turbidimetric assay.Results Compered with the control group,activity of T-SOD,Mn-SOD in CHD group were significantly decreased (t=9.41,6.34,all P<0.01).However,hs-CRP in CHD group were significantly increased to those in controls (t=3.09,P<0.05).The activity of T-SOD,Mn-SOD were negatively correlated with hs-CRP (P<0.01).Conclusion The variation of T-SOD,Mn-SOD activity and hs-CRP content were closely related to the occur-rence and development of CHD,they could be the impor tant indicators for riskfactors assessment of CHD.Moreover,con-joint analysis the correlation of T-SOD,Mn-SOD and hs-CRP has certain guiding significance for the clinical treatment and prognosis of coronary heart disease.

Effects of Manganese Superoxide Dismutase 9 Ala/Val Genetic Polymorphisms on Coronary Heart Disease / 现代检验医学杂志

Objective To study associations between manganese superoxide dismutase 9 Ala/Val (Mn-SOD 9 Ala/Val)genet-ic polymorphism and total superoxide dismutase (T-SOD)and Mn-SOD activity and the impact on coronary heart disease (CHD)were studied.Methods There were 82 CHD patients and 57 controls in this research.Sequencer was used to identify the genotype of Mn-SOD 9 Ala/Val genetic polymorphism and colorimeter was used to detect the serum T-SOD and Mn-SOD activity.Results Compared with the control group,the serum T-SOD and Mn-SOD activity of the CHD group was significantly reduced(t=4.83,6.57,P all<0.05),while the VV genotype and V allele of Mn-SOD 9 Ala/Val genetic poly-morphism of the CHD group were higher (χ2 =4.75,P <0.05).The serum T-SOD and Mn-SOD activity of the Mn-SOD 9 VV genotype was significantly lower than the Mn-SOD 9 AA genotype(t=2.96,3.11,P all<0.05).Conclusion The ser-um T-SOD and Mn-SOD activity in the CHD patients was reduced.Mn-SOD 9 Ala/Val genetic polymorphism was involved in the pathogenesis of CHD by influencing the Mn-SOD activity.

Extracellular superoxide dismutase, a potential extracellular biomarker candidate for prolactinoma / Superóxido dismutasa extracelular - candidato potencial a biomarcador extracelular del prolactinoma

AIM: To investigate whether the extracellular superoxide dismutase (EC-SOD) and manganese super-oxide dismutase (Mn-SOD) level changes during prolactinoma (PRL) development. METHODS: Surgical tissues from 37 female patients with PRL were tested for Mn-SOD and serum samples from such PRL patients were tested for EC-SOD level changes with Western Blot. The Mn-SOD level from blood cells was also investigated to show whether the Mn-SOD variation could locate tumorigenesis tissues. RESULTS: According to the patients' age analysis, age 20-40 years is high risk for getting PRL. There is a positive relationship between the PRL severity and EC-SOD. The Mn-SOD level from surgical tissues, but not blood cells, also shows a corresponding positive relationship to PRL severity, which indicates that elevated Mn-SOD might only happen in PRL tumorigenesis tissues. CONCLUSIONS: Extracellular superoxide dismutase is an extracellular protein and the serum EC-SOD could be a good candidate for the diagnoses of prolactinoma.

OBJETIVO: Investigar los cambios de niveles del superóxido dismutasa extracelular (EC-SOD) y el superóxido dismutasa de manganeso (Mn-SOD) durante el desarrollo del prolactinoma (PRL). MÉTODOS: Los tejidos quirúrgicos de 37 pacientes hembras con PRL fueron examinados para investigar los niveles de cambio de Mn-SOD, mediante la técnica de Western Blot. El nivel de Mn-SOD de las células sanguíneas fue investigado para ver si la variación de Mn-SOD puede indicar la localización de tejidos de tumorigénesis. RESULTADOS: Según el análisis de la edad de los pacientes, la edad de 20-40 años presenta un alto riesgo de desarrollar PRL. Hay una relación positiva entre la severidad del PRL y el EC-SOD. El nivel de Mn-SOD en los tejidos quirúrgicos - a diferencia de lo que ocurre en las células sanguíneas - muestra una relación positiva con respecto a la severidad del PRL, lo cual indica que un Mn-SOD elevado, sólo podría tener lugar en los tejidos de la tumorigénesis del PRL. CONCLUSIONES: El superóxido dismutasa extracelular (EC-SOD) es una proteína extracelular, y el EC-SOD sérico podría ser un buen candidato para diagnosticar el prolactinoma.

Association of Alanine-Valine Manganese Superoxide Dismutase Gene Polymorphism and Microheterogeneity Manganese Superoxide Dismutase Activity in Breast Cancer and Benign Breast Tissue / 한국유방암학회지

PURPOSE: Although the etiology of breast cancer is multifactorial, oxidative stress plays an important role in carcinogenesis. In this study, manganese superoxide dismutase (MnSOD) gene polymorphism and activity were evaluated in benign and breast cancer tissue. METHODS: One hundred and one females were enrolled in this study, 65 who were histopathologically diagnosed with breast cancer and 46 who were benign patients. MnSOD enzyme activity was determined using an indirect competitive inhibition assay and MnSOD gene polymorphism using poly merase chain reaction and agarose gel electrophoresis. RESULTS: MnSOD enzymatic activity (79.83+/-42.14) was lower in breast cancer tissue compared to benign tumors (236.18+/-46.37). At the same time, MnSOD enzymatic activity among Ala/Val patients was significantly lower in breast cancer tissue (39.19+/-7.33) than in Val/Val malignant breast tumors tissue (96.9+/-22.9). MnSOD enzymatic activity was significantly lower in Val/Val cancer tissue (96.9+/-22.9) than in benign tissue (255.44+/-42.7). CONCLUSION: Breast cancer tumors contain less MnSOD than benign breast samples. Patients with Ala/Val polymorphism had reduced MnSOD activity compared to patients with Val/Val breast cancer. Ala/Val gene polymorphism may be a risk factor associated with more advanced breast cancer stage. MnSOD gene polymorphism Ala/Val may be a risk factor associated with more advanced breast cancer stage, and reduction of MnSOD activity may be a mechanism of the progression of benign to malignant tumors. Further investigations are needed to evaluate the role of MnSOD in breast cancer progression.

Expression of manganese superoxide dismutase in rat blood, heart and brain during induced systemic hypoxia.

Background: Hypoxia results in an increased generation of ROS. Until now, little is known about the role of MnSOD - a major endogenous antioxidant enzyme - on the cell adaptation response against hypoxia. The aim of this study was to determine the MnSOD mRNA expression and levels of specific activity in blood, heart and brain of rats during induced systemic hypoxia. Methods: Twenty-five male Sprague Dawley rats were subjected to systemic hypoxia in an hypoxic chamber (at 8-10% O2) for 0, 1, 7, 14 and 21 days, respectively. The mRNA relative expression of MnSOD was analyzed using Real Time RT-PCR. MnSOD specific activity was determined using xanthine oxidase inhibition assay. Results: The MnSOD mRNA relative expression in rat blood and heart was decreased during early induced systemic hypoxia (day 1) and increased as hypoxia continued, whereas the mRNA expression in brain was increased since day 1 and reached its maximum level at day 7. The result of MnSOD specific activity during early systemic hypoxia was similar to the mRNA expression. Under very late hypoxic condition (day 21), MnSOD specific activity in blood, heart and brain was significantly decreased. We demonstrate a positive correlation between MnSOD mRNA expression and specific activity in these 3 tissues during day 0-14 of induced systemic hypoxia. Furthermore, mRNA expression and specific activity levels in heart strongly correlate with those in blood. Conclusion: The MnSOD expression at early and late phases of induced systemic hypoxia is distinctly regulated. The MnSOD expression in brain differs from that in blood and heart revealing that brain tissue can possibly survive better from induced systemic hypoxia than heart and blood. The determination of MnSOD expression in blood can be used to describe its expression in heart under systemic hypoxic condition.

Gene expression of manganese superoxide dismutase in human glioma cells.

Aim This study analyze the MnSOD gene expression as endogenous antioxidant in human glioma cells compared with leucocyte cells as control. Methods MnSOD gene expression of 20 glioma patients was analyzed by measuring the relative expression of mRNA and enzyme activity of MnSOD in brain and leucocyte cells. The relative expression of mRNA MnSOD was determined by using quantitative Real Time RT-PCR and the enzyme activity of MnSOD using biochemical kit assay (xantine oxidase inhibition). Statistic analysis for mRNA and enzyme activity of MnSOD was performed using Kruskal Wallis test. Results mRNA of MnSOD in glioma cells of 70 % sample was 0.015–0.627 lower, 10 % was 1.002-1.059 and 20 % was 1.409-6.915 higher than in leucocyte cells. Also the specific activity of MnSOD enzyme in glioma cells of 80 % sample showed 0,064-0,506 lower and 20 % sample was 1.249-2.718 higher than in leucocyte cells. Conclusion MnSOD gene expression in human glioma cells are significantly lower than its expression in leucocytes cells.

The Effect of Glucose Fluctuation on Apoptosis and Function of INS-1 Pancreatic Beta Cells / 당뇨병

BACKGROUND: Blood glucose level continuously fluctuates within a certain range in the human body. In diabetes patients, the extent of such fluctuation is large, despite the strict control of blood glucose. Blood glucose fluctuation has been shown to mediate more adverse effects on vascular endothelial cells and diabetes complications than chronic hyperglycemia, which has been explained as due to oxidative stress. As few previous studies have reported the effects of chronic and intermittent hyperglycemia on the apoptosis and function of pancreatic beta cells, this study reported herein was performed to investigate such effects on these cells. METHODS: For chronic hyperglycemia, INS-1 cells were cultured for 5 days with changes of RPMI 1640 medium containing 33 mM glucose every 12 hours. For intermittent hyperglycemia, the medium containing 11 mM glucose was exchanged with the medium containing 33 mM glucose every 12 hours. Apoptosis was assessed by TUNEL assay Hoechst staining and cleaved caspase 3. Insulin secretory capacity was assessed, and the expression of Mn-SOD and Bcl-2 was measured by Western blotting. RESULTS: In comparison to the control group, INS-1 cells exposed to chronic hyperglycemia and intermittent hyperglycemia showed an increase in apoptosis. The apoptosis of INS-1 cells exposed to intermittent hyperglycemia increased significantly more than the apoptosis of INS-1 cells exposed to chronic hyperglycemia. In comparison to the control group, the insulin secretory capacity in the two hyperglycemic states was decreased, and more with intermittent hyperglycemia than with chronic hyperglycemia. The expression of Mn-SOD and Bcl-2 increased more with chronic hyperglycemia than with intermittent hyperglycemia. CONCLUSION: Intermittent hyperglycemia induced a higher degree of apoptosis and decreased the insulin secretory capacity more in pancreatic beta cells than chronic hyperglycemia. This activity may be mediated by the anti-oxidative enzyme Mn-SOD and the anti-apoptotic signal Bcl-2.

Marternal Plasma ADMA Level, Placenta eNOS and Placenta Antioxidant Marker According to the Presence or Absence of Uterine Artery Notch on Color Doppler in the Third Trimester Pregnancies / 대한주산의학회잡지

PURPOSE: We examined pregnancy outcomes and maternal plasma asymmetric dimethylarginine (ADMA) concentrations in the presence or absence of uterine artery notch, and analyzed their relationships to the expression of placental endothelial nitric oxide synthase (eNOS) and antioxidant enzymes, including manganese superoxide dismutase (MnSOD), glutathione peroxidase (GPX), and catalase. METHODS: We assessed uterine artery doppler waveforms in 30 women who had been hospitalized for delivery. Plasma concentrations of ADMA were also measured. Tissue samples of placentas were obtained from 15 patients with diastolic notch and 15 patients without diastolic notch, according to uterine Doppler velocimetry analysis. We evaluated the placental expression of eNOS, MnSOD, GPX and catalase with Western blot analysis and eNOS with immunohistochemistry. RESULTS: The maternal plasma ADMA concentration increased significantly in the group with bilateral Uterine artery notch compared with the group without uterine artery notch (P=0.04). The expression of eNOS in the placenta significantly increased and the expression of MnSOD and GPX decreased significantly in the group with uterine artery notch at the third trimester. CONCLUSION: Uterine artery diastolic notch in pregnant women is associated with high maternal plasma ADMA, increased placental eNOS, and decreased MnSOD and GPX.

Association of manganese superoxide dismutase gene polymorphism (Ala-9Val) and breast cancer in males and females / Associação do polimorfismo do gene da superóxido dismutase dependente de manganês (Ala-9Val) em homens e mulheres com câncer de mama

INTRODUCTION: One of the several metabolic pathways involved in breast carcinogenesis is the human polymorphism in the mitochondrial targeting sequence Ala-9Val of the manganese superoxide dismutase (MnSOD) gene, which has been previously associated with increased risk of breast cancer in females. Since there is no previous report on this polymorphism in male breast cancer, the objective of this study is to analyze MnSOD polymorphism in a population of males and females with breast cancer from the southernmost state of Brazil, compared to healthy controls. METHODS: A case-control study of one hundred patients affected by breast cancer (11 men and 89 women) and 370 healthy age-adjusted database controls was performed. DNA was extracted from paraffin-embedded tumoral tissue. MnSOD polymorphism was determined by PCR-RFLP techniques using restriction enzyme Hae III. Chi-square test was used to compare MnSOD frequency distribution. RESULTS: MnSOD genotypic frequencies in all patients with breast cancer were AA = 15 percent; AV = 56 percent; VV = 29 percent and controls AA = 6.5 percent; AV = 68.1 percent and VV = 25.4 percent. Both male and female patients with breast cancer presented significantly higher AA frequencies compared to controls (p = 0.035), suggesting strong association of this genotype with breast cancer. A 2.15-fold (95 percent confidence interval [CI] 1.393-4.541) risk of breast cancer was found among individuals carrying the MnSOD AA allele-containing genotypes compared with the MnSOD VV and AV genotypes. DISCUSSION: These results confirm the already established association of MnSOD AA genotype with female breast cancer and further indicate a similar frequency distribution and increased risk in the male population.

INTRODUÇÃO: Uma das diversas rotas metabólicas envolvidas no processo de carcinogênese da mama é o polimorfismo Ala-9Val do gene da superóxido dismutase dependente de manganês, cuja associação com o aumento do risco de câncer de mama em mulheres já é bem estabelecida na literatura. Contudo, não existem estudos envolvendo esse polimorfismo no carcinoma de mama em homens, principalmente devido à baixa prevalência dessa neoplasia. O objetivo deste estudo é analisar o polimorfismo da MnSOD em uma população de homens e mulheres com câncer de mama no sul do Brasil, comparando tais achados com controles saudáveis. MÉTODOS: Foi realizado um estudo de caso-controle em cem pacientes com câncer de mama (11 homens e 89 mulheres) e 370 controles saudáveis. O DNA foi extraído do tecido tumoral emblocado em parafina. O polimorfismo da MnSOD foi determinado por técnicas de PCR-RFLP usando a enzima de restrição Hae III. O teste do qui quadrado foi usado para comparar a distribuição das freqüências dos polimorfismos. RESULTADOS: As freqüências genotípicas dos pacientes com câncer de mama foram AA = 15 por cento; AV = 56 por cento; VV = 29 por cento e dos controles AA = 6,5 por cento; AV = 68,1 por cento e VV = 25,4 por cento. Os pacientes com câncer de mama, tanto as mulheres como os homens, apresentaram freqüências significativamente mais elevadas do genótipo AA quando comparadas aos controles (p = 0,035), sugerindo associação forte desse genótipo com o câncer de mama. O intervalo de confiança foi de 1,393-4,541 (95 por cento) e o risco encontrado foi de 2,15 para indivíduos portadores do genótipo AA, quando comparados com os controles que tinham os genótipos VV e AV da MnSOD. DISCUSSÃO: Esses resultados confirmam a associação já estabelecida do genótipo da MnSOD AA com câncer de mama em mulheres e indicam distribuição de freqüência similar e risco aumentado na população masculina.

Effects of Repetitive Ischemic Preconditioning on the Expression of Cu,ZnSOD and MnSOD in Rectus Femoris Muscles of Rats / 체질인류학회지

The present study was performed to investigate the dynamics of Cu,ZnSOD and MnSOD expression following courses of reperfusion after repetitive ischemic preconditioning on the left rectus femoris muscle of Spraque-Dawley rats. Nine or thirty five weeks-old rats were subjected to three, six and ten cycles of ischemic preconditioning that was 5 min ischemia and 5 min reperfusion at the left common iliac artery. Left rectus femoris muscle was isolated 0, 3, 6, 24 and 72 hours of reperfusion after ischemic preconditioning and assayed by immunohistochemical staining with anti-Cu,ZnSOD and anti-MnSOD antibodies. The results were as follows; The immunoreactivities of Cu,ZnSOD and MnSOD were increased in the repectitive three and six cycles of ischemic preconditioning. However, after the repetitive ten cycles of ischemic preconditioning, the Cu,ZnSOD immunoreactivities were decreased in the nine weeks-old rats while MnSOD immunoreactivities were decreased in thirty five weeks-old rats. These findings suggest that severe damayes result from decrease of Cu,ZnSOD in nine weeks-old rats and decrease of MnSOD in thirty five weeks-old rats after ten cycles of ischemic preconditioning.

Placental Superoxide Dismutase Activity and Genetic Polymorphism in Preeclampsia and Normal Pregnancy / 대한주산의학회잡지

OBJECTIVE: Our purpose was to investigate urinary malondialdehyde (MDA), manganese superoxide dismutase (Mn-SOD) activity and polymorphism in placental tissues of women with preeclampsia and to evaluate oxidative stress in the pathophysiology of preeclampsia. METHODS: Urins and placental tissues were obtained from 20 normal and 20 preeclamptic women at 3rd trimester. Urinary MDA was assayed by an high performanance liquid chromatography (HPLC). The placental Mn-SOD activity was assayed by westen blotting and The placental Mn-SOD genotyping was assayed by PCR-RFLP. Data were analyzed statistically using Student's t-test and Chi-square test. RESULTS: 1) Urinary concentration of MDA was not significantly different in preeclampsia (4.43+/-2.37 ug/g) as compared with normotensive pregnancy (4.39+/-1.17 ug/g). 2) Preeclamptic women had similar Mn-SOD activity in placenta (1.04+/-0.04U/mL protein) as compared with normotensive pregnancy (1.44+/-0.34 U/mL protein). 3) No significant difference in the polymorphismthe of Mn-SOD genotype in placenta was observed between preeclampsia and normotensive pregnancy (X2=0.06, p>0.05) CONCLUSION: The findings in this study do not show that oxidative stress might be a pathogenetically relevent process causally contributing to the disease, and polymorphism in the Mn-SOD genotype in placenta do not seem to be risk factors for preeclampsia.