BACKGROUNDMany studies have suggested that cigarette smoking and polymorphisms of resistin and glutathione peroxidase-1 (GPx-1) genes are closely correlated with the pathogenesis of nonalcoholic fatty liver disease (NAFLD). However, few reports have investigated these associations with respect to NAFLD susceptibility. We, therefore, examined the distribution of polymorphisms in GPx-1 and resistin genes in NAFLD patients and healthy controls and analyzed the relationship between these polymorphisms and smoking status.

METHODSNine hundred NAFLD patients and 900 healthy controls were selected, and the genetic polymorphisms of resistin gene promoter-420C/G and GPx-1 gene Pro198Leu were analyzed by polymorphism-polymerase chain reaction (PCR) in DNA extracted from peripheral blood leukocytes. Interactions between the two mutants and the gene-environment interaction with cigarette smoking were also analyzed.

RESULTSGenotype frequencies of -420C/G (GG) and Pro198Leu (LL) were significantly higher in NAFLD cases (49.56% and 50.11%, respectively) compared with healthy controls (23.67% and 24.22%, respectively) (P = 0.0069; P = 0.0072). Moreover, the risk of NAFLD with -420C/G (GG) was significantly higher than in controls (odds ratio [OR] =3.1685, 95% confidence interval (CI) =1.9366-5.2073). Individuals carrying Pro198Leu (LL) had a high risk of NAFLD (OR = 3.1424, 95% CI = 1.7951-5.2367). Combined analysis of the polymorphisms showed that the -420C/G (GG)/Pro198Leu (LL) genotype was significantly more common in the NAFLD group than in the control group (39.44% vs. 12.78%, respectively, P = 0.0054), while individuals with -420C/G (GG)/Pro198Leu (LL) had a high risk of NAFLD (OR = 5.0357, 95% CI = 3.1852-7.8106). Moreover, the cigarette smoking rate in the NAFLD group was significantly higher than in the control group (OR = 1.8990, P = 0.0083 in the smoking index (SI) ≤400 subgroup; OR = 5.0937, P = 0.0051 in the SI >400 subgroup), and statistical analysis suggested a positive interaction between cigarette smoking and -420C/G (GG) (γ = 5.6018 in the SI ≤400 subgroup; γ = 4.4770 in the SI >400 subgroup) and Pro198Leu (LL) (γ = 5.7715 in the SI ≤400 subgroup; γ = 4.5985 in the SI >400 subgroup) in increasing the risk of NAFLD.

CONCLUSIONNAFLD risk factors include -420C/G (GG), Pro198Leu (LL) and cigarette smoking, and these three factors have a significant additive effect on NAFLD risk.

Female ; Genetic Predisposition to Disease ; genetics ; Glutathione Peroxidase ; genetics ; Humans ; Male ; Non-alcoholic Fatty Liver Disease ; metabolism ; Polymorphism, Single Nucleotide ; genetics ; Promoter Regions, Genetic ; genetics ; Resistin ; genetics ; Smoking ; genetics

OBJECTIVETo construct a Gpx1 and klk1 recombinant vector containing the kidney-specific promoter Ksp-cadherin.

METHODSHuman Gpx1, Klk1 and Ksp-cadherin cDNAs were amplified with PCR and inserted in a stepwise manner into the expressive vector pIRES-EGFP to construct the recombinant vector Ksp-cadherin-Gpx1-Klk1. The constructed vector was verified with restriction enzyme digestion and sequence analysis.

RESULTS AND CONCLUSIONThe recombinant expression vector Ksp-cadherin-Gpx1-Klk1 was constructed and identified successfully, which provides a potent tool for preparing transgenic animals to investigate gene therapy for ischemia-reperfusion injury in kidney transplantation.

Cadherins ; genetics ; Cloning, Molecular ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Glutathione Peroxidase ; genetics ; Humans ; Kallikreins ; genetics ; Kidney ; metabolism ; Promoter Regions, Genetic ; genetics

AIM: To investigate the antioxidant and anti-inflammatory effects of Shengmai San (SMS) and its ethyl acetate extract (SEa), n-butanol extract (SBu), and aqueous extract (SWe), and clarify the material base of SMS and the roles played by its fractions. METHODS: A mouse model of transient forebrain ischemia/reperfusion (I/R) by means of common carotid artery occlusion (CCAO) was used to investigate the effects of SMS and its three fractions. Histopathological damage, blood-brain barrier disruption, and antioxidant and inflammation-related parameters, including malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), myeloperoxidase (MPO), nitric oxide (NO), tumor necrosis factor-α (TNF-α) were measured. The chemical constituents of each fraction were identified by LC-MS. RESULTS: Eighteen lignans in SEa, and thirteen steroidal glycosides and ginsenosides in SBu were determined. SMS significantly inhibited I/R induced formation of histological injury and cerebral MPO activity. SMS showed the strongest antioxidant and anti-inflammatory effects against the I/R-caused injuries. SEa showed higher antioxidant activity than the other two fractions and SBu has a slightly stronger inhibition on the productions of NO and TNF-α. CONCLUSION SMS as a whole had the most effective protection against cerebral I/R-caused injuries compared with its fractions, which inferred that it contains different groups of compounds that contribute together to its protective effect.
Animals ; Chromatography, High Pressure Liquid ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Glutathione Peroxidase ; genetics ; metabolism ; Humans ; Male ; Malondialdehyde ; metabolism ; Nitric Acid ; metabolism ; Oxidative Stress ; drug effects ; Peroxidase ; genetics ; metabolism ; Protective Agents ; administration & dosage ; chemistry ; Rats ; Reperfusion Injury ; drug therapy ; genetics ; metabolism ; prevention & control ; Superoxide Dismutase ; genetics ; metabolism

OBJECTIVETo study the expression of WAVE1 and p22phox in peripheral blood mononuclear cells (PBMCs) in children with acute lymphocytic leukemia (ALL) and the relationship of WAVE1 with oxidative stress.

METHODSReal-time PCR was used for detecting WAVE1 and p22phox expression in PBMCs in 41 children with ALL and 10 normal controls. Plasma activity of superoxide dismutase (SOD) was measured by the xanthine oxidase method. Plasma activity of GSH-Px was measured by the DTNB reaction test.

RESULTSThe expression of WAVE1 and p22phox was significantly higher in the active ALL groups (newly diagnosed and relapse ALL) than that in the normal control and the complete remission (CR) ALL groups (<0.01). The CR ALL group showed increased WAVE1 and p22phox expression than those in the normal control group (<0.05). Plasma activities of SOD (22.62+/-7.39 U/mL) and GSH-Px (91.73+/-28.88 micromol/L) in the active ALL group were significantly lower than those in the normal control (166.35+/-27.93 U/mL and 490.94+/-39.38 micromol/L, respectively) and the CR ALL groups (107.11+/-28.57 U/mL and 267.56+/-82.64 micromol/L, respectively) (<0.01). WAVE1 expression was positively correlated with p22phox expression (r=0.34, <0.05) but negatively correlated with plasma activities of SOD and GSH-Px ( r=-0.336 and-0.408, respectively; <0.05).

CONCLUSIONSWAVE1 and p22phox expression in PBMCs increased and was associated with the disease course in children with ALL. Oxidative stress may be involved in the regulation of WAVE1 expression in ALL children.

Adolescent ; Child ; Child, Preschool ; Female ; Glutathione Peroxidase ; blood ; Humans ; Infant ; Leukocytes, Mononuclear ; metabolism ; Male ; NADPH Oxidases ; genetics ; Oxidative Stress ; Polymerase Chain Reaction ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; RNA, Messenger ; blood ; Superoxide Dismutase ; blood ; Wiskott-Aldrich Syndrome Protein Family ; genetics

OBJECTIVETo explore the role of oxidative stress and the antioxidant protein thioredoxin in the tumorigenesis and progression of gastric cancer.

METHODSThe plasma levels of adenosine deaminase (ADA), glutathione peroxidase (GPX), superoxide dismutase (SOD), and advanced oxidation protein products (AOPP) were determined by colorimetry, and the plasma levels of thioredoxin were determined by enzyme-linked immunosorbent assay (ELISA) in 48 gastric cancer patients and 30 healthy subjects. RT-PCR assay was employed to examine the expression levels of thioredoxin mRNA in the tissue samples of the patients.

RESULTSCompared with the healthy controls, patients with gastric cancer had significantly increased plasma levels of ADA and AOPP (P<0.05), decreased plasma GPX level (P<0.05), and similar plasma SOD levels. The plasma levels of thioredoxin were significantly higher in patients with gastric cancer than in the healthy controls (P<0.05). Thioredoxin levels was not associated with gender, age, degree of tumor cell differentiation, invasion depth, or lymph node metastasis (P>0.05), but was correlated to distant tumor metastasis (P<0.05). The expression of Trx mRNA was significantly higher in gastric carcinoma than in normal gastric tissue (P<0.05).

CONCLUSIONGastric cancer patients have high levels of oxidative stress and thioredoxin expression, and the latter is related to distant metastasis of the tumor.

Adenosine Deaminase ; blood ; Adult ; Advanced Oxidation Protein Products ; blood ; Aged ; Case-Control Studies ; Female ; Glutathione Peroxidase ; blood ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Oxidative Stress ; RNA, Messenger ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Superoxide Dismutase ; blood ; Thioredoxins ; genetics ; metabolism

OBJECTIVETo assess the association of blood selenium and polymorphism of glutathione peroxidase-1 (GPx-1) genes in patients with Keshan Disease (KD) and provide genetic evidence for KD susceptibility.

METHODSThe levels of whole blood selenium and the activity of GPx-1 were measured with spectrophotometric and enzymatic method among 71 KD patients and 290 controls (including 78 internal controls and 212 external controls). The genotype of GPx-1 at 198 site was analyzed by sequencing and PCR-RFLP. The functions of two GPx-1 variants were studied by rat neonatal cardiomyocytes transfection and expression plasmid.

RESULTSBlood level of selenium in KD patients was (0.8 ± 0.2) µmol/L, the internal controls' was (0.9 ± 0.2) µmol/L, and the external controls' was (1.2 ± 0.2) µmol/L (F = 4.888, P < 0.001).GPx-1 activity of KD patients was (73.0 ± 12.6) × 10(-10)U/RBC, internal controls' was (80.9 ± 9.2) × 10(-10)U/RBC, and external controls' was (115.8 ± 21.1) × 10(-10)U/RBC (F = 5.324, P < 0.001). Those of KD patients were significantly lower than controls. The polymorphism (Pro198Leu) of GPx-1 were identified; the frequency of Pro198Leu of KD patients was 21.1%, the frequency of controls was 10.7% (χ(2) = 5.588, P = 0.018). The level of blood selenium in variant subgroup (Pro198Leu or Leu198Leu) was (0.9 ± 0.2) µmol/L, and its in non-variant subgroup was (1.1 ± 0.3) µmol/L (t = 3.183, P < 0.01); The GPx-1 activity in variant subgroup was (86.1 ± 23.0) × 10(-10)U/RBC, and its in non-variant subgroup was (101.8 ± 25.9) × 10(-10)U/RBC (t = 5.784, P < 0.01). Further analysis revealed a synergistic-multiplicative interaction between presence of GPx-1 codon198 alleles and low blood selenium level. Over-expression of GPx-1 (198Leu) in rat cardiomyocytes caused 30% lower enzyme activity and less response to increasing concentrations of selenium than with over-expression of GPx-1 (198Pro).

CONCLUSIONLow blood selenium in carriers with the 198Leu-susceptible genotype of GPx-1 is associated with low GPx-1 activity, synergistic-multiplicative interaction was found between these two factors. And these two factors may increase the risk of KD.

Adult ; Animals ; Animals, Newborn ; Cardiomyopathies ; genetics ; Case-Control Studies ; Enterovirus Infections ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Glutathione Peroxidase ; genetics ; metabolism ; Humans ; Male ; Middle Aged ; Myocytes, Cardiac ; Polymorphism, Single Nucleotide ; Rats ; Selenium ; blood ; Transfection

OBJECTIVETo study the protective effect of recombinant adenovirus-mediated human cytosolic glutathione peroxidase (hCGPx) gene transfection on vascular endothelial cells ECV304 from oxidative damage.

METHODSpGEM-T Easy Vector containing hCGPx cDNA and recombinant adenovirus shuttle plasmid pACCMV-pLpA were used to construct the shuttle plasmid pACCMV-hCGPx for cotransfection of 293 cells with pJM17, thereby to obtain the recombinant adenovirus AdCMV-hCGPx. Cultured ECV304 cells were transfected with AdCMV-hCGPx for 24, 48 and 72 h, respectively, with the cells transfected with the empty vector serving as control, and hCGPx gene expression was then examined in the transfected cells. The transfected cell viability and apoptotic cell ratio were evaluated after treatment of the cells with H(2)O(2).

RESULTSThe expression ratio of hCGPx gene was significantly higher in the AdCMV-hCGPx-transfected cells than in those with empty vector transfection (P<0.01). The hCGPx gene-transfected cells showed significantly higher viability and significantly lower apoptotic ratio than the control cells following challenge with H(2)O(2)-induced oxidative damage.

CONCLUSIONhCGPx gene transfer mediated by recombinant adenovirus protects the vascular endothelial cells from oxidative damage in vitro, possibly due to the antioxidative and apoptosis-inhibiting effect of hCGPx.

Adenoviridae ; genetics ; Apoptosis ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Cytosol ; enzymology ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Flow Cytometry ; Genetic Vectors ; Glutathione Peroxidase ; biosynthesis ; genetics ; Humans ; Hydrogen Peroxide ; pharmacology ; Oxidative Stress ; Plasmids ; genetics ; Time Factors ; Transfection

Stress and emotion are associated with several illnesses from headaches to heart diseases and immune deficiencies to central nervous system. Terminalia arjuna has been referred as traditional Indian medicine for several ailments. The present study aimed to elucidate the effect of T. arjuna bark extract (TA) against picrotoxin-induced anxiety. Forty two male Balb/c mice were randomly divided into six experimental groups (n = 7): control, diazepam (1.5 mg·kg), picrotoxin (1 mg·kg) and three TA treatemt groups (25, 50, and 100 mg/kg). Behavioral paradigms and PCR studies were performed to determine the effect of TA against picrotoxin-induced anxiety. The results showed that TA supplementation increased locomotion towards open arm (EPM) and illuminated area (light-dark box test), and increased rearing frequency (open field test) in a dose dependent manner, compared to picrotoxin (P < 0.05). Furthermore, TA increased number of licks and shocks in Vogel's conflict. PCR studies showed an up-regulation of several genes, such as BDNF, IP, DL, CREB, GABA, SOD, GPx, and GR in TA administered groups. In conclusion, alcoholic extract of TA bark showed protective activity against picrotoxin in mice by modulation of genes related to synaptic plasticity, neurotransmitters, and antioxidant enzymes.
Animals ; Antioxidants ; metabolism ; Anxiety Disorders ; drug therapy ; genetics ; metabolism ; psychology ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Dopamine Agents ; administration & dosage ; GABA Agents ; administration & dosage ; Glutathione Peroxidase ; genetics ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Neuronal Plasticity ; drug effects ; Neurotransmitter Agents ; metabolism ; Phytotherapy ; Picrotoxin ; adverse effects ; Plant Bark ; chemistry ; Plant Extracts ; administration & dosage ; Serotonin Agents ; administration & dosage ; Superoxide Dismutase-1 ; genetics ; metabolism ; Terminalia ; chemistry

BACKGROUNDPioglitazone is effective in nonalcoholic steatohepatitis (NASH), but the mechanisms of action are not completely understood. This study was designed to investigate the effects of pioglitazone on hepatic nuclear factor-kappa B (NF-κB) and cyclooxygenases-2 (COX-2) expression in NASH rats.

METHODSThirty Sprague-Dawley male rats were randomly assigned to a control group (n = 10), NASH group (n = 10), and pioglitazone treatment group (n = 10). Liver tissues were processed for histology by hematoxylin & eosin and Masson stained. Serum alanine aminotransferase (ALT), cholesterol, triglyceride, fasting blood glucose (FBG), fasting insulin (FINS) levels and biochemical parameters of antioxidant enzyme activities, tumor necrosis factor alpha (TNF-α), prostaglandin E(2) (PGE(2)) levels in serum and liver were measured. The mRNA and protein expression of peroxisome proliferator-activated receptor gamma (PPARγ), NF-κB and COX-2 were determined by real-time polymerase chain reaction, Western blotting and immunohistochemistry. One-way analysis of variance (ANOVA) and Wilcoxon's signed-rank test was used for the statistical analysis.

RESULTSThere were severe steatosis, moderate inflammatory cellular infiltration and fibrosis in NASH rats. After pioglitazone treatment, steatosis, inflammation and fibrosis were significantly improved compared with the NASH group (χ(2) = 20.40, P < 0.001; χ(2) = 20.17, P < 0.001; χ(2) = 13.98, P = 0.002). Serum ALT, cholesterol, triglyceride, FBG, FINS levels were significantly elevated in the NASH group (P < 0.05). In the NASH group, total anti-oxidation competence (T-AOC), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) and malondialdehyde (MDA) levels in serum and liver were conspicuous disordered than those parameters in the control group. Meanwhile, TNF-α and PGE(2) levels in serum and liver were significantly increased compared with the control group. Immunohistochemistry showed NF-κB and COX-2 expression in liver was significantly elevated. However, PPAR? level was decreased in the NASH group. Real-time PCR and Western blotting revealed mRNA and protein expression of COX-2 were increased in the NASH group compared with the control group (0.57 ± 0.08 vs. 2.83 ± 0.24; 0.38 ± 0.03 vs. 1.00 ± 0.03, P < 0.001 and P = 0.004, respectively). After pioglitazone intervention, all of those parameters markedly improved (P < 0.05 or P < 0.01).

CONCLUSIONDown-regulating hepatic NF-κB and COX-2 expression, at least in part, is one of the possible therapeutic mechanisms of pioglitazone in NASH rats.

Alanine Transaminase ; blood ; metabolism ; Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Fatty Liver ; drug therapy ; metabolism ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; blood ; metabolism ; NF-kappa B ; genetics ; metabolism ; PPAR gamma ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Superoxide Dismutase ; metabolism ; Thiazolidinediones ; therapeutic use ; Tumor Necrosis Factor-alpha ; blood ; metabolism

OBJECTIVETo investigate the inhibitory effect of lycium pigment on lipopolysaccharide (LPS)-induced uveitis in rats and its mechanism.

METHODThe rat uveitis model was established by 30-day oral administration of lycium pigment (50, 100, 200 mg x kg(-1)) and footpad injection of LPS. Ocular tissues were collected for a histopathological inspection. The protein, nitric oxide and ADMA in aqueous humor, level of inducible nitric oxide synthase (iNOS) in retina, activities of serum total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX), and content of malondialdehyde (MDA) were determined by using Western blot, ELISA and biochemical methods.

RESULTAccording to the pathological study, lycium pigment (50, 100, 200 mg x kg(-1)) could notably reduce the inflammatory cell infiltration around corpus ciliare matrix of uveitis rats, and the concentration of protein and nitric oxide, and increased ADMA in aqueous humor. Lycium pigment (100, 200 mg x kg(-1)) could significantly inhibit the expression of iNOS in ocular tissues. In addition, lycium pigment (100, 200 mg x kg(-1)) also decrease the activities of serum T-AOC, SOD, GSH-PX, and the content of lipid peroxide MDA.

CONCLUSIONLycium pigment has the inhibitory effect on LPS-induced uveitis in rats. Its mechanism is related to the regulation of nitric oxide/ADMA pathway and the improvement of oxidation resistance.

Animals ; Glutathione Peroxidase ; genetics ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Lycium ; chemistry ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Pigments, Biological ; administration & dosage ; Plant Extracts ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; genetics ; metabolism ; Uveitis ; chemically induced ; genetics ; metabolism ; prevention & control