OBJECTIVE: To investigate the vascular damage effects and possible mechanism of acute exposure to ozone (O) in male Wistar rats. METHODS: One hundred and twenty male Wistar rats were randomly divided into six groups, 20 in each group. The experimental animals were placed in a gas poisoning cabinet, the control group was exposed to filtered air, and the treatment group was exposed to ozone at concentrations of 0.12 ppm, 0.5 ppm, 1.0 ppm, 2.0 ppm, and 4.0 ppm, respectively, for 4 hours. Arterial blood pressure data were obtained by PC-lab medical physiological signal acquisition system. Blood rheology indicators and blood biochemical indicators were detected by Tianjin Dean Diagnostic Laboratory. Serum endothelin-1 (ET-1), homocysteine (HCY), von Willebrand factor (vWF), 8-hydroxydeoxyguanosine (8-OhdG), interleukin (IL-6) and tumor necrosis factor alpha (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA) microplate assay. Oxidative stress indicators superoxide dismutase (SOD) activity and malondialdehyde (MDA) were determined by xanthine oxidase method, thiobarbituric acid (TBA) method, reduced glutathione (GSH) and nitric oxide (NO) were tested by using microplate colorimetry. Paraffin sections were prepared from thoracic aorta tissue, and vascular structure was observed by HE staining. RESULTS: Acute exposure to 0.12 ppm ozone could cause a significant increase in arterial systolic blood pressure (SBP). Exposure to different concentrations of ozone could cause a significant increase in plasma viscosity, and the K value of the ESR equation was significantly increased in the 1.0 ppm ozone exposure group. Both the relative and reduced viscosities were significantly reduced at ozone concentrations of 0.5 ppm and 4.0 ppm, while the red blood cell deformation index was increased significantly at ozone concentrations of 0.12 ppm, 0.5 ppm, 1.0 ppm, and 2.0 ppm. Acute ozone exposure resulted in the decrease of total cholesterol content. The content of high-density lipoprotein cholesterol (HDL-C) was significantly reduced in the 0.12 ppm ozone exposure group. When the ozone concentration was higher than 1.0 ppm, the body may also had an inflammatory reaction (increased TNF-α) and oxidative stress (increased MDA, decreased GSH). Acute exposure to ozone could lead to elevated levels of ET-1 in the blood, with significant differences in the 4.0 ppm concentration group, while HCY levels were decreased firstly and then increased, reaching the highest in the 1.0 ppm concentration group. No obvious pathological changes were observed in the thoracic aorta. CONCLUSION Acute ozone exposure can affect arterial blood pressure, blood rheology and cholesterol metabolism in rats. The possible mechanism is that ozone exposure leads to inflammatory reaction and oxidative stress reaction, causing vascular endothelial function damage, and vascular endothelial cells increase with ozone exposure concentration.
Animals ; Blood Vessels ; injuries ; Deoxyguanosine ; analogs & derivatives ; blood ; Endothelin-1 ; blood ; Homocysteine ; blood ; Interleukin-6 ; blood ; Male ; Malondialdehyde ; analysis ; Oxidative Stress ; Ozone ; toxicity ; Rats ; Rats, Wistar ; Superoxide Dismutase ; analysis ; Tumor Necrosis Factor-alpha ; blood ; von Willebrand Factor ; analysis

PURPOSE: To compare oxidative stress status in the aqueous humor of highly myopic eyes and control eyes. METHODS: Aqueous humor samples were collected from 15 highly myopic eyes (high myopia group) and 23 cataractous eyes (control group) during cataract surgery. Central corneal thickness, corneal endothelial cell density, hexagonality of corneal endothelial cells, and cell area of corneal endothelial cells were measured using specular microscopy. Axial length was measured using ultrasound biometry. 8-Hydroxydeoxyguanosine (8-OHdG) and malondialdehyde levels were measured using enzyme-linked immunosorbent assay. RESULTS: 8-OHdG level was lower in the aqueous humor of myopic patients than in that of control group (p = 0.014) and was positively correlated with central corneal thickness and negatively correlated with axial length (r = 0.511, p = 0.02; r = -0.382, p < 0.001). There was no correlation between 8-OHdG level and corneal endothelial cell density, hexagonality, or cell area. Malondialdehyde level did not show any correlation with any parameters evaluated. CONCLUSIONS: 8-OHdG might be a sensitive biomarker for evaluating oxidative stress status in the eye. Oxidative stress level was lower in the aqueous humor of highly myopic eyes compared to that in control eyes, which indicates lower metabolic activity in these eyes.
Aged ; Aqueous Humor/*metabolism ; Deoxyguanosine/*analogs & derivatives/metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Malondialdehyde/*metabolism ; Middle Aged ; Myopia/*metabolism/physiopathology ; *Oxidative Stress ; Refraction, Ocular/*physiology ; Severity of Illness Index

To explore the protective effect of Angelica sinensis polysaccharides(ASP) on subacute renal damages induced by D-galactose in mice and its mechanism. Male C57BL/6J mice were randomly divided into 3 groups, with 10 mice in each group. The D-galactose model group was subcutaneously injected with D-galactose (120 mg x kg(-1)), qd x 42; the ASP + D-galactose model group was intraperitoneally injected with ASP since the 8th day of the replication of the D-galactose model, qd x 35; and the normal control group was subcutaneously injected with saline at the same dose and time. On the 2nd day of after the injection, the peripheral blood was collected to measure the content of BUN, Crea, UA, Cys-C; paraffin sections were made to observe the renal histomorphology by HE staining; senescence-associated β-g-alactosidase (SA-β-Gal) stain was used to observe the relative optical density (ROD) in renal tissues; transmission electron microscopy was assayed to observe the renal ultrastructure; the renal tissue homogenate was prepared to measure the content of SOD, GSH-PX, MDA; the content of AGEs and 8-OH-dG were measured by ELISA. According to the result, compared with the D-galactose model group, the ASP + D-galactose model group showed obviously decreases in the content of BUN, Crea, UA, Cysc, AGES, 8-OH-dG, the number of hardening renal corpuscle, renal capsular space and renal tubular lumen, ROD of SA-β-Gal staining positive kidney cells, mesangial cells, basement membrane thickness, podocyte secondary processes fusion and MDA and increases in the number of normal renal corpuscle, ribosome and rough endoplasmic reticulum in podocytes, the activity of SOD and GSH-PX. In Conclusion, A. sinensis polysaccharides can antagonize kidney subacute damages induced by D-galactose in mice. Its protective mechanism may be correlated with the inhibition of the oxidative stress injury.
Angelica sinensis ; chemistry ; Animals ; Deoxyguanosine ; analogs & derivatives ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Galactose ; adverse effects ; Humans ; Kidney ; anatomy & histology ; drug effects ; injuries ; Kidney Diseases ; chemically induced ; drug therapy ; metabolism ; prevention & control ; Male ; Mice ; Mice, Inbred C57BL ; Oxidative Stress ; drug effects ; Polysaccharides ; administration & dosage ; Protective Agents ; administration & dosage

OBJECTIVETo study the protective effect of Sanhuangyinchi Fang drug serum (SF) against hydrogen peroxide-mediated DNA oxidative damage in LO2 cells.

METHODSThe LO2 cells were randomly divided into the control group, H(2)O(2) group, SF groups (5%, 10%, and 15%) and vitE group. The morphological features of the treated LO2 cells were observed under inverted microscope. The viability of the treated cells was assessed with CCK-8 method, and the activity of SOD, CAT and GSH-PX were detected biochemically. Reactive oxygen species (ROS) levels, the content of 8-OHdG, and DNA damage of the cells were evaluated by flow cytometry, ELISA, and Comet assay, respectively.

RESULTSCompared with H(2)O(2) group, the cells in SF groups (10% and 15%) and vitE group showed higher cell survival rate (P<0.05) and higher SOD, CAT, GSH-PX (P<0.05) and ROS scavenging activities (P<0.01) with markedly decreases the content of 8-OHdG (P<0.01) and reduced tailing ratio, tail length, tail moment and Olive tail moment (P<0.05).

CONCLUSIONSF drug serum, especially at the concentration of 15%, can protect LO2 cells from H(2)O(2)-mediated DNA oxidative damage.

Cell Line ; Comet Assay ; DNA Damage ; Deoxyguanosine ; analogs & derivatives ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Hydrogen Peroxide ; toxicity ; Oxidation-Reduction ; Oxidative Stress ; Protective Agents ; pharmacology ; Reactive Oxygen Species

Parkinson's disease (PD) is an age-related progressive neurodegenerative disease associated with selective loss of dopaminergic neurons. The characteristic hallmark of the disease is intracytoplasmic proteinacious inclusion bodies called Lewy bodies, primarily consisting of a presynaptic protein alpha-synuclein. Oxidative stress-mediated damage to macromolecules have been shown to occur frequently in PD. Oxidative damage to DNA in the form of oxidized guanine (8-oxodG) accumulates in both the mitochondrial and nuclear DNA of dopaminergic neurons of the substantia nigra in PD. 8-oxodG-mediated transcriptional mutagenesis has been shown to have the potential to alter phenotype of cells through production of mutant pool of proteins. This review comprehensively summarizes the role of oxidative stress-mediated damage incurred during neurodegeneration, and highlights the scope of transcriptional mutagenesis event in leading to alpha-synuclein aggregation as seen in PD.
Amino Acid Sequence ; Animals ; Deoxyguanosine/*analogs & derivatives/metabolism ; Humans ; Molecular Sequence Data ; Mutagenesis ; *Oxidative Stress ; Parkinson Disease/*genetics/metabolism/pathology ; Protein Aggregation, Pathological/*genetics/metabolism/pathology ; Substantia Nigra/metabolism/*pathology ; Transcription, Genetic ; alpha-Synuclein/chemistry/*genetics

INTRODUCTIONOxidative stress, assessed using 8-hydroxy-2'-deoxyguanosine (8-OHdG), can be associated with arterial stiffness in patients with type 2 diabetes mellitus (T2DM) and/or hypertension (HT). We investigated the correlation between urinary 8-OHdG and pulse wave velocity (PWV) in hypertensive and non-hypertensive T2DM patients with fair glycaemic control to determine the clinical significance of HT as a comorbidity in the diabetic state.

METHODSClinical data, including traditional cardiovascular risk factors, diabetic complications, prescribed agents, urinary 8-OHdG level and brachial-ankle PWV, was collected from T2DM patients with and without HT.

RESULTSThere were 76 patients (45 men, 31 women; mean age 61 years; mean haemoglobin A1c level 6.5%) in the study cohort. T2DM patients with HT had significantly higher mean PWV than patients without HT (1,597 cm/s vs 1,442 cm/s; p < 0.05). Patients with HT showed no significant difference in 8-OHdG levels relative to those without HT (median 7.9 ng/mg creatinine vs 8.8 ng/mg creatinine; p > 0.05). Simple linear correlation and stepwise multiple linear regression analyses revealed that 8-OHdG levels correlated independently, significantly and positively with PWV among T2DM patients with HT (r = 0.33, p < 0.05; β= 0.23, p < 0.05). No significant correlation was observed between 8-OHdG levels and PWV among T2DM patients without HT.

CONCLUSIONIn the hypertensive state, oxidative stress can be responsible for the development of arterial stiffness, even in patients with fairly well controlled T2DM. Oxidative stress management may be necessary for the prevention of cardiovascular disease in this population.

Adult ; Aged ; Aged, 80 and over ; Ankle Brachial Index ; Cohort Studies ; Deoxyguanosine ; analogs & derivatives ; urine ; Diabetes Complications ; urine ; Diabetes Mellitus, Type 2 ; urine ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Hypertension ; complications ; urine ; Male ; Middle Aged ; Oxidative Stress ; Pulse Wave Analysis ; Vascular Stiffness

OBJECTIVETo establish a method for simultaneously determining the urinary concentrations of 8-hydroxy-2'-deoxyguanosine (8-OHdG), trans, trans-muconic acid (tt-MA), and S-phenylmercapturic acid (S-PMA) in subjects exposed to benzene.

METHODSAfter being purified by a solid-phase extraction column, the urine samples were transferred to a liquid chromatography-mass spectrometry system, and the concentrations of 8-OHdG, tt-MA, and S-PMA were determined by external standard method. A C18 reversed-phase column was used as the chromatographic column, and methanol/acidic ammonium formate solution was used as the mobile phase for gradient elution. The mass spectrometer was operated in a multi-reaction monitoring mode.

RESULTSFor tt-MA, the calibration curves were linear in the range of 10-1000 µg/L, and the recovery rates were over 90% (relative standard deviation (RSD) < 3%) at spiked levels of 50 µg/L and 500 µg/L. For S-PMA and 8-OHdG, the calibration curves were linear in the range of 1-100 µg/L, and the recovery rates were over 85% (RSD < 5%) at spiked levels of 5 µg/L and 50 µg/L.

CONCLUSIONThis determination method meets the requirement of Biological materials-

METHODSof monitoring-Guide of development (WS/T 68-1996) and can be used for simultaneous determination of 8-OHdG, tt-MA, and S-PMA in urine.

Acetylcysteine ; analogs & derivatives ; urine ; Benzene ; poisoning ; Chromatography, Liquid ; methods ; Deoxyguanosine ; analogs & derivatives ; urine ; Humans ; Mass Spectrometry ; Occupational Exposure ; prevention & control ; Sorbic Acid ; analogs & derivatives ; metabolism

OBJECTIVETo study the potential association of DNA oxidation and DNA methylation, in vitro cultured cells were exposed to different doses of H2O2, 8-oxo-dG formation, cell DNA 5-mC contents were analyzed to explore the time- dose-response relationship of DNA oxidation and DNA methylation.

METHODSA549 cells were exposed to different doses of H2O2, 8-oxo-dG formation and cell genomic DNA 5-mC contents were analyzed by a high-performance liquid chromatography system and high performance capillary electrophoresis (HPCE), respectively.

RESULTSH2O2 induced the formation of 8-oxo-dG and 5-mC in different characteristics, it need at least 10 days for significant changes in the level of DNA methylation, whereas under the same conditions, changes in the level of DNA oxidation cast only 12 hours. H2O2 induced decreased levels of DNA methylation in A549 cells in a dose-dependent manner. In a certain range of time and dose, it showed a negative correlation between DNA oxidation and DNA methylation.

CONCLUSIONThe study suggests that oxidative DNA could lead to reduced levels of DNA methylation, DNA oxidation may affect the regulation of cellular methylation mechanisms, in the course of chemical mutagenesis, DNA oxidation may be an earlier important molecule event than DNA methylation.

Cell Line ; DNA ; chemistry ; DNA Damage ; DNA Methylation ; Deoxyguanosine ; analogs & derivatives ; chemistry ; Humans ; Hydrogen Peroxide ; toxicity ; Oxidative Stress

OBJECTIVETo explore the effects of hydrogen-rich saline (HS) on liver of severely scalded rats with delayed resuscitation.

METHODSTwenty-four SD rats were inflicted with 40% TBSA full-thickness scald using a temperature-controlled scalding apparatus. The injured rats were divided into lactated Ringer's solution (RS) and HS groups according to the random number table, with 12 rats in each group. Rats in groups RS and HS were respectively resuscitated with an intraperitoneal injection of 4 mL × kg⁻¹ × %TBSA⁻¹ of RS or HS (self-prepared, with concentration of hydrogen 0.6 mmol/L) 6 hours after injury up to 48 hours after scald. The infusion volume of the second 24 hours after injury was a half of that of the first 24 hours. At post scald hour (PSH) 6 (before resuscitation), 12, 24, and 48, blood was collected from the heart of 3 rats in each group, and then the rats were sacrificed for harvesting liver tissue. The pathological change in liver tissue was observed with HE staining. The number of hepatic neutrophils was counted with a hematocytometer. Serum levels of AST and ALT were determined with full-automatic biochemical analyzer. Contents of TNF-α, IL-1β, IL-6, and 8-hydroxy-2'-deoxyguanosine (8-OHdG) in liver tissue were determined with ELISA. Absorbance value of malondialdehyde (MDA) in liver tissue was detected and quantified with spectrophotometer. Data were processed with analysis of variance of repeated measurement and LSD-t test.

RESULTSAt PSH 48, moderate infiltration of inflammatory cells and hepatic hyperemia were observed in rats of group HS as compared with group RS. At PSH 12, 24, and 48, the number of neutrophils in group HS was respectively (25.3 ± 1.8) × 10⁵, (19.6 ± 0.6) × 10⁵, and (14.1 ± 3.2) × 10⁵ cells per mililitre, and they were significantly lower than those in group RS \[(31.9 ± 2.0) × 10⁵, (30.9 ± 2.2) × 10⁵, and (23.8 ± 3.0) × 10⁵ cells per mililitre, with t values respectively 5.6, 7.6, and 8.7, P values below 0.05\]. At PSH 6 and 12, the serum levels of AST and ALT and the levels of TNF-α, IL-1β, and IL-6 in liver tissue were close between the two groups (with t values respectively 0.3-3.9 and 0.9-3.8, P values above 0.05). At PSH 24 and 48, the serum levels of AST and ALT in group HS were respectively (308 ± 24) and (210 ± 15) U/L and (93 ± 7) and (70 ± 5) U/L, which were significantly lower than those in group RS \[(541 ± 39) and (505 ± 18) U/L, with t values respectively 17.5 and 16.7, P values below 0.05; (156 ± 9) and (166 ± 21) U/L, with t values respectively 30.3 and 6.9, P values below 0.05\]. At PSH 24 and 48, the levels of TNF-α, IL-1β, and IL-6 in liver tissue in group HS were respectively (20.7 ± 1.6) and (13.7 ± 1.5) pg/mg, (7.7 ± 1.5) and (6.3 ± 1.2) pg/mg, and (8.7 ± 1.2) and (6.0 ± 2.0) pg/mg, which were significantly lower than those in group RS \[(32.7 ± 5.0) and (25.7 ± 4.0) pg/mg, with t values respectively 5.2 and 5.7, P values below 0.05; (16.3 ± 2.5) and (12.0 ± 2.7) pg/mg, with t values both as 4.7, P values below 0.05; (14.7 ± 2.1) and (13.3 ± 1.5) pg/mg, with t values respectively 10.4 and 4.4, P values below 0.05\]. The level of MDA at PSH 6 and levels of 8-OHdG at PSH 6 and 12 in liver tissue were close between the two groups (with t values respectively 0.1, 0.7, and 4.3, P values above 0.05). In group HS, the levels of MDA in liver tissue at PSH 12, 24, and 48 were respectively (15.3 ± 1.5), (8.7 ± 1.2), and (6.7 ± 1.5) mmol/mg, and the levels of hepatic 8-OHdG at PSH 24 and 48 were respectively (124 ± 12) and (79 ± 10) pg/mg, which were significantly lower than those in group RS \[(27.3 ± 4.7), (20.3 ± 1.5), and (14.0 ± 1.0) mmol/mg, with t values respectively 5.2, 5.7, and 5.1, P values below 0.05; (191 ± 10) and (136 ± 15) pg/mg, with t values respectively 8.0 and 8.1, P values below 0.05\].

CONCLUSIONSResuscitation with HS could protect liver of severely scalded rats with delayed resuscitation possibly by reducing infiltration of neutrophils, thus lowering the content of inflammatory cytokines, and effectively alleviating oxidative stress.

Animals ; Burns ; metabolism ; physiopathology ; Deoxyguanosine ; analogs & derivatives ; blood ; Hydrogen ; pharmacology ; Interleukin-6 ; Liver ; drug effects ; metabolism ; physiopathology ; Male ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Resuscitation ; Sodium Chloride ; pharmacology ; Soft Tissue Injuries ; Time Factors ; Tumor Necrosis Factor-alpha ; blood

Cigarette smoke is associated with the development of several diseases, such as chronic obstructive pulmonary disease (COPD). The purpose of this study was to investigate genotoxicity and heat shock protein 70 (Hsp70) in human airway smooth muscle cells (HASMCs) exposed to cigarette smoke extract (CSE). HASMCs was exposed to CSE with different doses for 24 h. The level of 8-hydroxydeoxyguanosine (8-OHdG) was determined by using HPLC-ECD, the DNA damage was analyzed by using comet assay, and apoptosis was examined by using Annexin-FITC/PI staining. The production of Hsp70 after CSE stimulation was tested. Results indicated that CSE significantly increased the level of 8-OHdG, DNA damage and cell apoptosis, and reduced the production of Hsp70. In particular, levels of Hsp70 were inversely correlated with 8-OHdG, DNA damage and cell apoptosis. It was concluded that cigarette smoke induced genotoxicity and decreased the production of cell protective protein Hsp70, which may contribute to the development of some airway diseases.
Apoptosis ; DNA Damage ; Deoxyguanosine ; analogs & derivatives ; metabolism ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Lung ; cytology ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Smoke ; adverse effects ; Tobacco ; toxicity ; Tumor Cells, Cultured