ABSTRACT
Objective To investigate the effect of the fusion of leader peptide on the structure of human manganese superoxide dismutase (SOD2) and anti-cisplatin (DDP)-induced renal injury. Methods The effect of mitochondrion targeting sequence (MTS) on the structure and activity of SOD2 was analyzed by structure prediction and superoxide dismutase (SOD) specific-activity determination. The DDP injury model of Kunming (KM) mice was established, and amifostine (AMFT) was set as a positive control. Indicators such as kidney index, renal function, kidney antioxidant capacity, and appearance and pathology changes of mice kidney were used to evaluate the effect of MTS-SOD2 against DDP-induced kidney injury. Results The MTS leader peptide seemed to change the secondary and tertiary structures of SOD2 to some extent, but it also increased the specific activity of the MTS-SOD2 protein. Pre-administration of a medium dose of MTS-SOD2 (0.84 mg/kg) before the use of DDP significantly reduced the level of renal malondialdehyde and increased the SOD activity and total antioxidant capacity (T-AOC) in the kidney, thereby reducing the renal pathological damage and consequently maintaining renal function. The overall protective effect of MTS-SOD2 was comparable to or even better than that of 200 mg/kg AMFT. Conclusion The MTS leader peptide enhances the activity of SOD2 and confers it with an excellent anti-DDP-induced renal-injury effect because of its transmembrane function.
ABSTRACT
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><sub>1-42</sub> （A<italic>β</italic><sub>1-42</sub>， 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<sup>-1</sup>·d<sup>-1</sup>）， Kaixinsan low dose， medium dose and high dose groups received corresponding doses of Kaixinsan （10， 20， 40 g·kg<sup>-1</sup>·d<sup>-1</sup>， 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.
ABSTRACT
BACKGROUND: Excessive exercises cause a large accumulation of oxidative active substances in the body to damage skeletal muscle cells. Mitochondria play a key role in energy metabolism during exercise. Studies have shown that Rhodiola can reduce the level of lipid peroxidation in muscle tissue and protect damaged endothelial cells. OBJECTIVE: To explore the mechanism underlying Rhodiola improving skeletal muscle function of mice with high intensity exercise by regulating mitochondrial function. METHODS: The study protocol was approved by the Ethics Committee of Xi’an Shiyou University in China. Forty BALB/c mice were divided into blank control group, exercise group, Rhodiola control group and Rhodiola intervention group. Mice in the blank control had no exercise and intervention. Mice in exercise group were given intragastric administration of normal saline followed by high intensity exercise. Mice in Rhodiola intervention group and Rhodiola control group were given intragastric administration of the mixture of Rhodiola and normal saline, followed by exercise or not. The interventions were performed once a day for 28 consecutive days. Body mass, forearm grip strength and exhaustion time were observed. Western blot assay was used to detect expression of manganese superoxide dismutase protein, p53 protein, mitochondrial origin and autophagy-associated protein in the skeletal muscle. RT-qPCR was used to detect skeletal muscle Mfn-1, Mfn-2, Opa-1, Drp-1, and fis-1 mRNA expression. RESULTS AND CONCLUSION: (1) From the 2nd week, the grip strength of forelimbs in the exercise group was significantly lower than that in the other three groups (P 0.05). (2) At the 3rd and 4th weeks, the exhaustion time of weight-bearing swimming training was significantly shorter in the exercise group than the Rhodiola intervention group (P 0.05). Compared with the blank control group, the Rhodiola exercise intervention group also showed a downward trend in the expression of fusion gene and an upward trend in the expression of Drp-1 mRNA, but there was no significant difference between the two groups (P > 0.05). To conclude, Rhodiola can significantly improve the exercise endurance of mice with high intensity exercise, which may be related to the improvement of skeletal muscle mitochondrial autophagy, origin and fusion-division.
ABSTRACT
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.
ABSTRACT
AIM: To investigate the effects of manganese superoxide dismutase mimic (MnSODm) on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS) induced ulcerative colitis (UC) in rats and to probe into its underlying mechanism. METHODS: Wistar rats were randomly divided into blank group, model group, sulfasalazine (SASP, 500 mg/kg) group, and different doses of MnSODm (10, 20 and 40 mg/kg) groups. Ulcerative colitis was induced in rats by rectal administration of 100 mg/kg TNBS dissolved in 50% ethanol. Rats were killed after SASP and different doses of MnSODm treatment 7 days. The disease activity index (DAI) was recorded, and then the colonic injury and inflammation were assessed by the colon weight/length ratio and microscopic damage scores. The serum and colon tissues activities myeloperoxidase (MPO) were detected by biochemistry method. The activities of glutathione peroxidase (GSH-Px), inducible nitric oxide synthase (iNOS), and the levels of glutathione (GSH) and NO in colon tissues were also detected. The levels of TNF-α, IL-4 and IL-10 in the colon tissues were measure by ELISA. Western blot was undertaken to determine the phosphorylation levels of AKT and PI3K. RESULTS: Compared with the model group, the colonic weight/length ratios, microscopic damage scores and colon tissues and serum MPO activity were significantly decreased in MnSODm groups (P<0.05 or P<0.01). INOS, NO, TNF-α, PI3K, p-AKT levels in colon tissues were also significantly decreased in MnSODm treatment groups; while the activity of GSH-Px and the concentration of GSH, IL-4 and IL-10 obviously increased (P<0.05, P<0.01). CONCLUSION: MnSODm is protective against colitis via antioxidant activity and by inhibiting inflammatory mediators and then down-regulating PI3K/AKT signaling pathways.
ABSTRACT
Objective Dangerous placenta previa(PPP) combined with placenta implantation seriously threatens maternal life safety. This paper aim to explore the changes of MnSOD and SIRT3,the expression of SIRT3 in maternal placenta PPP combined with placenta implantation, and the relationship between trophoblast invasion and placental implantation. Methods 90 cases with placenta implantation of pernicious placenta previa were collected from January 2014 to June 2018 in Anhui Maternal and Child Health Hospital. According to the depth of placental villus invading uterine myometrium, 30 cases of placenta adhesion, 30 cases of placental implantation, and 30 cases of placenta penetration, 30 cases of normal control group．Immunohistochemical SP and Western blot were used to detect the expression of MnSOD and SIRT3 in placental tissues of the study group and the control group, then compared and analyzed. Results Compared with the control group, the expression of MnSOD and SIRT3 in the placental implantation group were increased. With the increasing of placental implantation degree, the level of MnSOD and SIRT3 decreased significantly (P<0.05). Western blot showed that , the relative protein expressions of MnSOD/β-actin and SIRT3/β-actin in the control group were (0.39±0.05) and (0.41±0.08), which were higher than those in the adhesion group[(0.35±0.04), (0.32±0.02)], the implantion group[(0.28±0.02), (0.20±0.03)], and the penetration group[(0.23±0.01), (0.17±0.02)]. The difference was statistically significant(P<0.05). Conclusion The expressions of MnSOD and SIRT3 incytoplasm or nucleus of invasive trophoblasts and placental tissues of pregnant women with placental implantation is significantly decreased, both of which are involved in the occurrence and development of placental implantation, but the specific pathogenesis still needs to be further explored.
ABSTRACT
Objective To investigate the role of epigenetic regulations of peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) in the development of diabetic retinopathy and the metabolic memory phenomenon after hyperglycemia was terminated.Methods Diabetic rat model was established by intraperitoneal injection of streptozotocin (STZ).Sixty diabetic rats were randomly divided into 3 groups,poor glycemic control group rats were maintained in poor glycemic control for 4 months;semi glycemic control group rats were maintained in poor glycemic control for 2 months,followed by good glycemic control for 2 additional months;good glycemic control group rats were maintained in good glycemic control for 4 months.Twenty normal rats served as control group.The mRNA expression of PGC-1α and superoxide dismutase 2 (SOD2) of retina were measured by real-time PCR;the expression of PGC-1α and manganese superoxide dismutase (MnSOD) protein were measured by Western blot;the situation of DNA methylation in the promotor region of PPARGC1A was measured by bisulfite sequencing.Results The body-weight in the control group was significantly higher than that in the poor glycemic control group,semi glycemic control group and good glycemic control group (all at P =0.000).The blood glucose value in the poor glycemic control group was significantly higher than that in the control group (P =0.000).The expression levels of PGC-1 α mRNA were significantly lower and the expression levels of SOD2 mRNA were significantly higher in the good glycemic control group,semi glycemic control group and poor glycemic control group than those in the control group (all at P＜0.05).The expression levels of PGC-1α and SOD2 mRNA were significantly different between the good glycemic control group and poor glycemic control group (both at P＜0.05).Compared with the control group,the expression levels of PGC-1α and MnSOD protein were decreased in the diabetic model groups,with significant differences between them (all at P＜0.05).The expression level of PGC-1 α protein was significantly higher in the good glycemic control group than that in the poor glycemic control group (P＜0.05).Diabetes increased DNA methylation in the promotor region of PPARGC1A gene of retina.The DNA methylation level was significantly higher in the poor glycemic control group and semi glycemic control group than that in the control group (P =0.008,0.031).No statistical difference was found between the poor glycemic control group and semi glycemic control group (P ＞ 0.05).Conclusions The expressions of PGC-1o mRNA and protein and MnSOD protein in the retina of STZ induced diabetic rats are decreased,the expression of SOD2 mRNA is increased,the expression changes have metabolic memory characteristics.Increased DNA methylation in the promotor region of PPARGC1A when exposed to high glucose may have a role in the regulation of PGC-1 α expression and metabolic memory.
ABSTRACT
Reactive oxygen is closely related to the occurrence and development of tumors. Manganese superoxide dismutase (MnSOD),as an antioxidant enzyme that scavenges superoxide anion in the mitochondrial matrix,plays an important role in maintai-ning mitochondrial redox homeostasis and protecting mitochondrial DNA. Many studies have discovered that MnSOD has different ex-pression levels in different tumors,and has both anti-cancer and cancer-promoting functions,but its functional conversion mecha-nism remains unclear. This article intends to investigate the role of MnSOD in tumorigenesis and development,and to review the main regulatory mechanisms of its expression level and activity.
ABSTRACT
OBJECTIVE: To investigate the effect of the power-frequency electromagnetic field on antioxidant indexes in operation personnel. METHODS: By random sampling method,a total of 58 workers who work in 500 kV transformer substations for at least 5. 0 years were selected as the exposure group,and 60 administrative staffs in the same enterprise were included as the control group. The power frequency electromagnetic fields in the 2 groups were measured. The cumulative exposure dose of individual magnetic field was calculated. Peripheral venous blood was collected in these 2groups and was examined with the superoxide dismutase( SOD) activity,malondialdehyde( MDA) level and the relative gene expression level of manganese superoxide dismutase( MnSOD). RESULTS: There was no statistical significance in the SOD activity,MDA level and the relative gene expression level of MnSOD between the exposure group and the control group( P > 0. 05). The logistic regression analysis results indicated that cumulative exposure dose was not correlated with the SOD activity,MDA level and the relative gene expression level of MnSOD after adjusting confounding factors such as age,smoking,alcohol and tea drinking( P > 0. 05). CONCLUSION: Power-frequency electromagnetic field exposure has no significant effects on the SOD activity,MDA level and the relative gene expression level of MnSOD of operation workers.
ABSTRACT
Objective To investigate the relationship between manganese superoxide dismutase (Mn-SOD)9 Ala/Val genetic polymorphisms and the levels of blood lipid and homocysteine (HCY).Methods The genotypes of Mn-SOD 9 Ala/Val ge-netic polymorphisms were identified by sequencing method,the serum activities of T-SOD and Mn-SOD were detected by colorimetric method,the serum level of HCY was detected by enzymatic method,and the serum levels of cholesterol (TC), triglyceride (TG),high density lipoprotein cholesterol (HDL-C)and low density lipoprotein cholesterol (LDL-C)were de-tected by end-point method in 137 patients with coronary heart disease (CHD)and 85 controls.Results Compared with the control group,the VV genotype and V allele of Mn-SOD 9 Ala/Val genetic polymorphisms in the CHD group were higher, while the serum activities of T-SOD and Mn-SOD in the CHD group was significantly lower.The serum activities of T-SOD and Mn-SOD of the Mn-SOD 9 VV genotype was significantly lower than the Mn-SOD 9 AA genotype.Compared with the Mn-SOD 9 AA genotype,the serum levels of TC,TG,LDL-C and HCY of the Mn-SOD 9 VV genotype were significantly higher,while the serum level of HDL-C was significantly lower.The serum activity of Mn-SOD was negativelycorrelated with the serum levels of TC,TG,LDL-C and HCY and positively correlated with the serum level of HDL-C.Conclusion The antioxidative ability in patients with CHD was decreased.Mn-SOD 9 Ala/Val genetic polymorphisms led to lipid metab-olism disorders by affecting the Mn-SOD activity,promoting the development of CHD.HCY resulted in increased oxidative substances by self-oxidation and inhibition of the Mn-SOD activity,increasing the risk of CHD.
ABSTRACT
Enhanced oxidative stress is a hallmark of cisplatin nephrotoxicity, and inhibition of poly(ADP-ribose) polymerase 1 (PARP1) attenuates oxidative stress during cisplatin nephrotoxicity; however, the precise mechanisms behind its action remain elusive. Here, using an in vitro model of cisplatin-induced injury to human kidney proximal tubular cells, we demonstrated that the protective effect of PARP1 inhibition on oxidative stress is associated with sirtuin 3 (SIRT3) activation. Exposure to 400 µM cisplatin for 8 hours in cells decreased activity and expression of manganese superoxide dismutase (MnSOD), catalase, glutathione peroxidase (GPX), and SIRT3, while it increased their lysine acetylation. However, treatment with 1 µM PJ34 hydrochloride, a potent PARP1 inhibitor, restored activity and/or expression in those antioxidant enzymes, decreased lysine acetylation of those enzymes, and improved SIRT3 expression and activity in the cisplatin-injured cells. Using transfection with SIRT3 double nickase plasmids, SIRT3-deficient cells given cisplatin did not show the ameliorable effect of PARP1 inhibition on lysine acetylation and activity of antioxidant enzymes, including MnSOD, catalase and GPX. Furthermore, SIRT3 deficiency in cisplatin-injured cells prevented PARP1 inhibition-induced increase in forkhead box O3a transcriptional activity, and upregulation of MnSOD and catalase. Finally, loss of SIRT3 in cisplatin-exposed cells removed the protective effect of PARP1 inhibition against oxidative stress, represented by the concentration of lipid hydroperoxide and 8-hydroxy-2'-deoxyguanosine; and necrotic cell death represented by a percentage of propidium iodide–positively stained cells. Taken together, these results indicate that PARP1 inhibition protects kidney proximal tubular cells against oxidative stress through SIRT3 activation during cisplatin nephrotoxicity.
Subject(s)
Humans , Acetylation , Catalase , Cell Death , Cisplatin , Deoxyribonuclease I , Down-Regulation , Glutathione Peroxidase , In Vitro Techniques , Kidney , Lipid Peroxides , Lysine , Oxidative Stress , Plasmids , Poly Adenosine Diphosphate Ribose , Poly(ADP-ribose) Polymerases , Propidium , Sirtuin 3 , Superoxide Dismutase , Transfection , Up-RegulationABSTRACT
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.
ABSTRACT
<p><b>OBJECTIVE</b>The roles of cerebrovascular oxidative stress in vascular functional remodeling have been described in hindlimb-unweighting (HU) rats. However, the underlying mechanism remains to be established.</p><p><b>METHODS</b>We investigated the generation of vascular reactive oxygen species (ROS), Nox2/Nox4 protein and mRNA levels, NADPH oxidase activity, and manganese superoxide dismutase (MnSOD) and glutathione peroxidase-1 (GPx-1) mRNA levels in cerebral and mesenteric smooth muscle cells (VSMCs) of HU rats.</p><p><b>RESULTS</b>ROS production increased in cerebral but not in mesenteric VSMCs of HU rats compared with those in control rats. Nox2 and Nox4 protein and mRNA levels were increased significantly but MnSOD/GPx-1 mRNA levels decreased in HU rat cerebral arteries but not in mesenteric arteries. NADPH oxidases were activated significantly more in cerebral but not in mesenteric arteries of HU rats. NADPH oxidase inhibition with apocynin attenuated cerebrovascular ROS production and partially restored Nox2/Nox4 protein and mRNA levels, NADPH oxidase activity, and MnSOD/GPx-1 mRNA levels in cerebral VSMCs of HU rats.</p><p><b>CONCLUSION</b>These results suggest that vascular NADPH oxidases regulate cerebrovascular redox status and participate in vascular oxidative stress injury during simulated microgravit.</p>
Subject(s)
Animals , Male , Acetophenones , Cerebral Arteries , Metabolism , Glutathione Peroxidase , Metabolism , Hindlimb Suspension , Membrane Glycoproteins , Metabolism , Mesenteric Arteries , Metabolism , Myocytes, Smooth Muscle , Metabolism , NADPH Oxidase 2 , NADPH Oxidase 4 , NADPH Oxidases , Metabolism , Rats, Sprague-Dawley , Reactive Oxygen Species , Superoxide Dismutase , MetabolismABSTRACT
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.
Subject(s)
Adult , Female , Humans , Middle Aged , Young Adult , Pituitary Neoplasms/metabolism , Prolactinoma/metabolism , Superoxide Dismutase/metabolism , Biomarkers, Tumor/metabolism , Blood Cells/metabolism , Blotting, Western , Case-Control Studies , Severity of Illness IndexABSTRACT
Objective: To explore the effect of manganese superoxide dismutase (MnSOD) overexpression on the proliferation of esophageal cancer cell line TE-1 in vivo and in vitro . Methods: TE-1 cells were transfected with a plasmid binding MnSOD cDNA, and then TE-1Mm cells (with moderate expression of MnSOD) and TE-1Mh cells (with high expression of MnSOD) were established. RT-PCR and Western blotting were used to detect the mRNA and protein expressions of target MnSOD gene in both of TE-1Mm and TE-1Mh cells, respectively. The ability of cell proliferation and apoptosis and the cell cycle distribution were examined by plate colony-forming test and flow cytometry (FCM). The xenograft tumor models in nude mice were established. The effect of MnSOD overexpression on cell proliferation was evaluated in vivo , and the expression of MnSOD protein in xenograft tumors was detected by immunohistochemistry and Western blotting. Results: TE-1 cells with different expression levels of MnSOD proteins were established by transfection with different amounts of plasmids binding MnSOD cDNA. The colony-formation rates of TE-1Mm and TE-1Mh cells were (23.0±2.7)% and (45.3±4.5)%, respectively, which were both significantly higher than those in TE-1 cells (34.7±4.2)% and TE-1n cells (33.7±4.7)%, P<0.05. The apoptosis rate of TE-1Mm (10.6±1.0)% was significantly higher than those in TE-1 cells (34.7±4.2)% and TE-1n cells (33.7±4.7)%, and the apoptosis rate of TE-1Mh (10.6±1.0)% was significantly lower than those in TE-1 cells and TE-1n cells (P<0.05). FCM revealed that the percentage of TE-1Mh cells was decreased in G 0/G1 phase and increased in G2/M and S phases, while the percentage of TE-1Mm cells was increased in G0/G 1 phase and decreased in G2/M and S phases induced by MnSOD overexpression. The growth of xenograft tumors was inhibited in TE-1Mm cell-implanted group, while which was improved in TE-1Mh cell-implanted group. The expressions of MnSOD protein in TE-1Mm and TE-1Mh cells were significantly higher than those in TE-1 and TE-1n cells (P<0.05). Conclusion: MnSOD overexpression exerts a two-way effect involving inhibition or promotion on the proliferation of TE-1 cells in vivo and in vitro.
ABSTRACT
Objective To investigate the expression of MnSOD and E-cadhefin in nasopharyngeal carcinoma(NPC) tissue and its relationship with clinicopathological features and prognosis. Methods The expression of MnSOD and E-cadherin were detected by immunohistochemistry method in 60 NPC patients. Results Of the whole group,lymph node positive group and lymph node negative group,the strong positive rate of MnSOD protein was 47% (28/60) ,49% (25/51 patients) and 33% (3/9) (x2 =0.76,P =0.382), respectively.The corresponding strong positive rate of E-cadherin protein was 47% (28/60) ,43% (22/51) and 78% (7/9) (x2 =3.69,P =0.047) ,respectively.The expression of MnSOD increased with T stage and N stage.The higher expression of MnSOD was significantly associated with the larger size of metastatic lymph node(r =0.46 ,P =0.002) ,more radioresistance and poorer prognosis,but not with the region of lymph node metastasis(r =0.223,P = 0.116).The lower expression of E-cadherin was closely relevant with higher N stage and the smaller region of lymph node metastasis(r =-0.33,P = 0.020),but not with T stage,lymph node size or radiosensitivity(r =-2.19,P=0.093;r=-0.07,P=0.623;r=-0.18,P=0.170).Multi variate analysis showed that MnSOD and E-canherin were independent prognostic factors (x2= 4.45,P = 0.035;x2 =5.12,P=0.024). Conclusions High expression of MnSOD may stimulate tumor growth and reduce radiosensitivity.High expression of E-cadherin may inhibit lymphatic metastasis,while has no rela tionship with tumor growth and invasion.MnSOD and E-cadherin could affect the prognosis of NPC patients.
ABSTRACT
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.
Subject(s)
Female , Humans , Pregnancy , Arginine , Blotting, Western , Catalase , Glutathione Peroxidase , Nitric Oxide Synthase Type III , Placenta , Plasma , Pregnancy Outcome , Pregnancy Trimester, Third , Pregnant Women , Rheology , Superoxide Dismutase , Uterine ArteryABSTRACT
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.
ABSTRACT
AIM: To investigate the transfection efficiency of mouse liver with non-viral vector containing manganese superoxide dismutase (Mn-SOD) gene. METHODS: The eukaryotic expression vector, gWiz/Mn-SOD, encoding human manganese superoxide dismutase was constructed. The plasmids of gWiz/Mn-SOD were mixed with cationic lipids, followed by injection into mice via branch of superior mesenteric vein, to induce Mn-SOD over-expression in murine liver detected by RT-PCR, Western blotting, SOD activity and immunohistochemical staining. RESULTS: gWiz/Mn-SOD transfection resulted in the obvious expression of exogenous Mn-SOD mRNA and protein in hepatic tissues at 8 hours after injection, and elevated mitochondria SOD activity 8.4 times in transfected hepatocytes than that in non-transfected cells at 72 hours after injection. It was showed that nearly 70% of mouse hepatocytes was obviously Mn-SOD positive after transfection. CONCLUSION: High expression efficiency of Mn-SOD gene in mouse liver is achieved safely by injection of gWiz/Mn-SOD and cationic lipid mixture into branch of superior mesenteric vein. [