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

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.

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.

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.

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.

Correlation of Gene Polymorphism of Manganese Superoxide Dismutase and Type 2 Diabetic Retinopathy / 上海第二医科大学学报

0.05).The frequency of VV genotype of Mn-SOD in the DR(+) group was significantly higher than that in the DR(-) group(92.0% vs 77.1%,P

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.

Screening of differential expression proteins from human oral lichen planus and normal oral mucosa by two-dimensional difference gel electrophoresis and mass spectrometry / 实用口腔医学杂志

Objective:To find biomarkers for oral lichen planus by comparing differential expressing proteins. Methods:10 cases of oral lichen planus and normal oral mucosa tissues were collected.Total protein was extracted; differential proteome profiles were established and analyzed by two-dimensional polyacrylamide gel electrophoresis(2D-PAGE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Results:(1)The well-resolved,reproducible 2-DE patterns of oral lichen planus and normal oral mucosa were obtained. The results showed that average protein spots were 1 576?67 and 1 608?73 in oral lichen planus and normal oral mucosa respectively, (2) The 13 differential protein spots were identified by Imaging Master 2D image analysis software between oral lichen planus and normal oral mucosa. There were 7 protein spots in oral lichen planus were higher than those in normal oral mucosa, 6 protein spots in oral lichen planus were lower than those in normal oral mucosa. 10 differential expressing proteins were analyzed by mass spectrometry and bioinformation. 4 of them were well characterized including manganese superoxide dismutase (Mn-SOD), Annexin I, vimentin and unknown proteins. Conclusion:Differential expression proteins might be candidate biomarkers for diagnosis of oral lichen planus;and proteomic technique is valuable for screening the diagnostic biomarkers.

Induction of Metallothionein and Manganese-Containing Superoxide Dismutase by Paraquat in Peripheral White Blood Cells of Patients with Gastric Cancer

PURPOSE: It is known that deficiency of metallothionein (MT) and manganese-containing superoxide dismutase (Mn-SOD), scavengers of reactive oxygen species, results in aging and carcinogenesis by inducing DNA damage. Paraquat can produce reactive oxygen species and induce antioxidants in human. In this study, an attempt was made to verify the relation between gastric carcinogensis and the induction rates of these antioxidants. METHODS: Peripheral blood of 24 randomly selected patients with gastric cancer, who were treated at Chosun University Hospital between Febuuary 1999 and December 1999, was examined. 3 male and 3 female patients in each decade from 40 to 70 years were selected. Twenty-four (24) volunteers with no laboratory, chemical, radiologic and endoscopic abnormalities during the same period were used for the normal control group. White blood cells were isolated from peripheral blood and incubated in culture media, including paraquat, for 18 hours at 37oC. MT and Mn-SOD mRNA expressions were examined using the reverse transcription-polymerase chain reaction (RT-PCR) method. RESULTS: The induction rates of MT and Mn-SOD mRNA in the cancer group due to paraquat were lower than those in the control group. Also, the rates decreased in both groups with age. CONCLUSION: The inducibilities of MT and Mn-SOD mRNA by paraquat may play a role in the carcinogenesis of gastric cancer and in the aging process. Based on this result, patients with a high risk of gastric cancer should be screened actively for early detection.

Anti-oxidative effect of total extract of astragalus(TEA) on aging mice / 中国临床药理学与治疗学

Aim To study the mechanism of the anti-aging effect of total extract of astragalus(TEA) in mice. Methods MDA content, the activities of Mn-SOD and GSHpx and the reduced to oxidized glutathione (GSH/GSSG) ratio in mitochondria of D-galactose(D-gal) treated mice and 17-month-old mice were measured. Results Treatment with TEA(40 mg?kg-1?d-1 ig) for 10 wk would lower the content of MDA and restore activities of Mn-SOD,GSHpx and GSH/GSSG ratio in mitochondria of D-gal treated mice.Treatment with TEA (40 mg?kg-1 ?d-1 ig) for 3 mon had the same effect on 17-month-old mice.Conclusion TEA has anti-aging effect on D-gal treated mice and 17-month-old mice significantly, probably being related to its anti-oxidative effect.

Effects of Cadmium on Superoxide Radical, Superoxide Dismutase, Catalase and ATPase Activity in Liver, Kidney and Testicle of Rats in Vtm and in Vivo / 예방의학회지

Production of free radicals of superoxide anion in tissues by cadmium, activities of superoxide dismutase and catalase to protect tissue damages caused by the free radicals and ATPase that plays an important role in energy metabolism at cellular level were investigated. Experiments in viro were conducted with liver, kidney and testicle tissue homogenates of rats adding 0.05~0.50 mM cadmium chloride, and in vivo experiments administering single dose of 5mg of cadmium/kg of body weight in 0.1% cadmium chloride solution intraperitoneally 48 hours prior to evisceration. Production of superoxide radicals in liver and testicle increased with addition of cadmium in vitro, but not in kidney. In vivo experiments, however superoxide radicals slightly increased in liver and kidney but not in testicle. Superoxide dismutase (Cu, Zn-SOD and Mn-SOD), catalase and ATPase (total, (Mg++)- & (Na+)- (K+)-) activity decreased in the presence of cadmium in dose dependent manner. Reduction of these enzyme activities varied not only with dosage of cadmium but also with type of tissue and between in vitro and in vivo experiment.