Cardiovascular Diseases ; prevention & control ; Humans ; Thioredoxins

BACKGROUND AND OBJECTIVES: Oxidative stress is thought to play important role in cardiovascular disease. Thioredoxin is an important biomarker for determining the degree of oxidative stress. However, the relationship between the plasma thioredoxin levels and myocardial damage has not been investigated. SUBJECTS AND METHODS: We measured the plasma thioredoxin levels in the patients suffering with acute myocardial infarction and who also underwent successful primary angioplasty. We then compared the plasma thioredoxin levels and the clinical parameters in acute myocardial infarction patients (n=37) in order to examine the relationship between oxidative stress and myocardial damage. RESULTS: The plasma thioredoxin level was significantly related with the initial WBC count (r=0.349, p<0.05) and the myocardial damage, the peak CK level (r=0.489, p<0.01), the CK increment (r=0.452, p<0.05), the peak MB level (r=0.417, p<0.05), and the MB increment (r=0.364, p<0.05). We divided the patients into two groups according to the plasma thioredoxin levels. There was a significant difference in myocardial damage between the low and high plasma thoiredoxin levels at the initial WBC count (10174.2+/-3380.4/uL vs 13500+/-3740.7/uL, respectively; p<0.01) and the cardiac enzyme, the peak CK level (2565.2+/-1389.9 IU/L vs 4045.9+/-1978.9 IU/L, respectively; p=0.02), the CK increment (2309.6+/-1351.8 IU/L vs 3762.8+/-2079.7 IU/L, respectively; p=0.03), the peak MB level (208.7+/-127.5 IU/L vs 322.7+/-146.3 IU/L, respectively; p=0.02), and the MB increment (173.8+/-128.4 IU/L vs 277.7+/-158.9 IU/L, respectively; p=0.05). CONCLUSION: High thioredoxin levels were associated with the degree of oxidative stress and the extent of myocardial damage. Thioredoxin levels may be used as a new surrogate biomarker for the severity of oxidative stress and the extent of myocardial damage in the patients suffering with acute myocardial infarction.
Angioplasty* ; Cardiovascular Diseases ; Humans ; Myocardial Infarction* ; Oxidative Stress ; Plasma* ; Thioredoxins*

Animals ; Lung/metabolism* ; Rats ; Silicon Dioxide/toxicity* ; Thioredoxins/metabolism*

PURPOSE: Thioredoxin is an endogenous antioxidant. It regulates the activities of transcriptional factors such as NF-kB(nuclear factor kappa B)and AP-1(activator protein-1) and it increases the synthesis of cytokines, preventing cellular proliferation and apoptosis. The aim of this study was to clarify the role of thioredoxin on apoptosis-inducing neuronal cell injury. We investigated the protective effects of thioredoxin against apoptosis-inducing neuronal cell injury through intracellular mechanism by 6-hydroxydopamine and serum deprivation. METHODS: PC 12 cells were cultured in RPMI 1640 media containing 10% fetal calf serum and subcultured in 96-well plates. Each well contained 30,000 cells. Cells were treated with hydrogen peroxide, diamide or 6-hydroxydopamine 30 minutes after thioredoxin treatment and then incubated for 24 hours. Cytotoxicity and cellular viability were assessed by measuring lactate dehydrogenase(LDH) release and MTT reduction. RESULTS: Thioredoxin increased cytotoxicity of PC cells treated with 6-hydroxydopamine by increasing LDH release and decreasing MTT reduction. In the serum deprivation condition, thioredoxin increased cytotoxicity of PC cells by increasing LDH release. CONCLUSION: Thioredoxin potentiates oxidative injury through intracellular mechanisms by 6-hydroxydopamine and serum deprivation instead of protecting. The cytotoxicity of thioredoxin may be mediated by decreasing the activity of NF-kB, which has been reported recently to protect against cellular apoptosis. Evidence suppors that the cytotoxic effect was not increased in the presence of serum in this study. Therefore, we found that the antioxidant effects of thioredoxin depended on mechanisms of injuries.
Antioxidants ; Apoptosis ; Cell Proliferation ; Cytokines ; Diamide ; Hydrogen Peroxide ; Lactic Acid ; Neurons* ; NF-kappa B ; Oxidopamine ; Thioredoxins*

Apoptosis signal-regulating kinase 1 (ASK1), a mitogen- activated protein kinase kinase kinase, plays pivotal roles in stress responses. In addition, ASK1 has emerged as a key regulator of immune responses elicited by pathogen-associated molecular patterns (PAMPs) and endogenous danger signals. Recent studies have demonstrated that reactive oxygen species (ROS)-dependent activation of ASK1 is required for LPS-stimulated cytokine production as well as extracellular ATP-induced apoptosis in immune cells. The mechanism of ROS-dependent regulation of ASK1 activity by thioredoxin and TRAFs has been well characterized. In this review, we focus on the molecular details of the activation of ASK1 and its involvement in innate immunity.
Apoptosis ; Immunity, Innate ; MAP Kinase Kinase Kinase 5 ; Oxygen ; Phosphotransferases ; Protein Kinases ; Reactive Oxygen Species ; Thioredoxins

Apoptosis signal-regulating kinase 1 (ASK1), a mitogen- activated protein kinase kinase kinase, plays pivotal roles in stress responses. In addition, ASK1 has emerged as a key regulator of immune responses elicited by pathogen-associated molecular patterns (PAMPs) and endogenous danger signals. Recent studies have demonstrated that reactive oxygen species (ROS)-dependent activation of ASK1 is required for LPS-stimulated cytokine production as well as extracellular ATP-induced apoptosis in immune cells. The mechanism of ROS-dependent regulation of ASK1 activity by thioredoxin and TRAFs has been well characterized. In this review, we focus on the molecular details of the activation of ASK1 and its involvement in innate immunity.
Apoptosis ; Immunity, Innate ; MAP Kinase Kinase Kinase 5 ; Oxygen ; Phosphotransferases ; Protein Kinases ; Reactive Oxygen Species ; Thioredoxins

BACKGROUND AND OBJECTIVES: This study was designed to examine the effects of diamide and thioredoxin (TRX) on vascular endothelial cells in order to clarify the mechanism by which vascular damage is mediated by oxygen free radicals. MATERIALS AND METHODS: The pulmonary artery endothelial cell (PAEC) line derived from bovine serum was cultured for 8 hours in media supplemented with various concentrations of diamide and TRX. The XTT assay, MTS assay, SRB assay, LDH activity and lipid peroxidation tests were perfomed. RESULTS: In XTT and MTS assays, diamide significantly decreased the cell viability of cultured PAEC in a dose- and time-dependent manner. Diamide showed a decrease in the amount of total protein, although it showed an increase of lipid peroxidation and LDH activity in cultured PAEC. In regards to the protective effect of TRX on diamide-induced cytotoxicity, this showed an increase of total protein, however it showed a decrease of lipid peroxidation and LDH activity. CONCLUSION: Our results suggest that diamide has a vasculotoxic effect on cultured bovine PAEC and that TRX is very effective in the protection of diamide-induced cytotoxicity by duye to the increase of total protein and the decrease of lipid peroxidation and LDH activity in these cultures.
Cell Survival ; Diamide* ; Endothelial Cells* ; Endothelium, Vascular ; Free Radicals ; Lipid Peroxidation ; Oxygen ; Pulmonary Artery ; Thioredoxins*

The objective of this study was to construct an improved thioredoxin fusion protein expression system, and express the cathelicidin-derived peptide, Lf-CATH2. The improved fusion vector Lf-CATH2-pET32α(-TS) was successfully constructed by firstly deleting the thrombin site and S tag from the pET-32α vector, then inserting the Lf-CATH2 plus a thrombin site instead. Afterwards, Lf-CATH2 was expressed in Escherichia coli as fusion protein. After the cleavage by thrombin, Lf-CATH2 was released and subsequently separated using affinity chromatography. The antimicrobial activity of purified Lf-CATH2 was also examined. The improved expression vector significantly increased enzyme cleavage efficiency by 37%, and Lf-CATH2 could be expressed in high yield and maintain the biological activity. This novel thioredoxin fusion protein expression system enables a quick production of high-yield bioactive cationic peptides like cathelicidins.
Cathelicidins ; biosynthesis ; Chromatography, Affinity ; Escherichia coli ; Genetic Vectors ; Recombinant Fusion Proteins ; biosynthesis ; Thioredoxins ; genetics

PURPOSE: Thioredoxin is an endogenous antioxidant which directly scavenges reactive oxygen species(ROS) and regenerates oxidatively damaged protein by reducing potential at the redox active disulfide(-Cys-Gly-Pro-Cys-) site. Under oxidative stress, thiredoxin plays a protective and adaptative role by inducing expressions. The aim of this study was to clarify the role of thioredoxin on oxidative neuronal cell injury. We investigated the protective effects of E. coli thioredoxin, also acting as a substrate for mammalian thioredoxin reductase, against oxidative neuronal cell injury under oxidative stresses such as hydrogen peroxide and diamide. METHODS: PC 12 cells were cultured in RPMI 1640 media containing 10% fetal calf serum and subcultured in 96-well plates. Each well contained 30,000 cells. Cells were treated with hydrogen peroxide or diamide 30 minutes after thioredoxin treatment and then incubated for 24 hours. Cytotoxicity and cellular viability were assessed by measuring of lactate dehydrogenase(LDH) release and MTT reduction. RESULTS: Thioredoxin not only decreased the cytotoxicity of PC 12 cell treated with hydrogen peroxide by decreasing LDH release and preventing the decrease of MTT reduction but also thioredoxin showed greater protective effects when simultaneously treated with hydrogen peroxide. Also, thioredoxin decreased cytotoxicity by decreasing LDH release from PC 12 cells damaged by diamide. Thioredoxin did not prevent the decrease of MTT reduction on PC 12 cells damaged by diamide. CONCLUSION: Thioredoxin protected PC 12 cells under oxidative stresses by directly scavenging and inhibiting oxidants such as hydrogen peroxide and diamide.
Diamide ; Hydrogen Peroxide ; Lactic Acid ; Neurons* ; Oxidants ; Oxidation-Reduction ; Oxidative Stress ; Oxygen ; Thioredoxin-Disulfide Reductase ; Thioredoxins*

To investigate the role of thioredoxin interacting protein(TXNIP)/ nucleotides-binding oligomerization domain-like receptor protein(NLRP)3 inflammasome in the sciatic nerve of streptozotocin(STZ)-induced diabetic rats. The diabetic rat model was established by single intraperitoneal injection of STZ.The rats with matched sex and age were taken as normal control group.The blood glucose and body weight were monitored.The mechanical withdrawal threshold was measured by von Frey filaments at 12 weeks after the model was established.At 12 weeks,the rats were sacrificed and the sciatic nerves were separated for Luxol fast blue staining,the expressions of TXNIP,NLRP3,caspase-1,and interleukin(IL)-1β were detected by immunohistochemistry and Western blot method,and the levels of IL-1β and IL-18 in serum were measured by enzyme-linked immunosorbent assay(ELISA). The expression of TXNIP protein in the sciatic nerve of diabetic rats was 3.78±0.08,which significantly increased than that in the normal control group(0.99±0.06)(=26.980,<0.0001).Compared with the normal control group(0.97±0.05),the expression of NLRP3 protein in the diabetic group(2.44±0.16)was significantly higher(=8.885,<0.0001).The expression of cleaved caspase-1 was 4.45±0.19 in the diabetic group and 1.08±0.06 in the normal control group,and the difference was significant(=16.900,<0.0001).The expression of IL-1β protein in the diabetic group(4.50±0.16)was significantly higher than that(1.19±0.08)in the normal control group(=18.630,<0.0001).Compared with the normal control group,the levels of IL-1β [(110.50±8.80)pg/ml (17.97±3.18)pg/ml,=9.892,<0.0001] and IL-18 [(591.70±8.78)pg/ml (160.70±8.33)pg/ml,=35.620,<0.0001] in the serum of diabetic rats significantly increased. The pathogenesis of diabetic peripheral neuropathy may be related to increased expression of TXNIP,activation of NLRP3 inflammasome,and downstream inflammation,which may provide a new target for diabetic peripheral neuropathy therapy.
Animals ; Diabetes Mellitus, Experimental ; Inflammasomes ; Nucleotides ; Rats ; Sciatic Nerve ; Streptozocin ; Thioredoxins