OBJECTIVETo investigate the apoptosis-inducing effect of trichloroethylene (TCE) on cultured normal human epidermis keratinocytes (NHEK) in vitro.

METHODSNR(50) values (the concentration of neutral red absorbed is reduced to 50%) of TCE on NHEK were assayed by neutral red uptake (NRU), and the administered dose of TCE was determined. Lipid peroxidation (LPO) and oxidative stress were assessed by measurement of malondialdehyde (MDA) contens and superoxide dismutase (SOD) activity. Transmission electron microscope (TEM) were used to observe morphologic changes, flow cytometer (FCM) was used to measure DNA contents and calculate cell apoptosis rate and proliferation index (PI).

RESULTSNR(50) values of TCE on NHEK was found to be 4.53 mmol/L (95% CI: 3.92-5.13 mmol/L). The increase in MDA content and inhibition of SOD activity in a concentration-dependent manner were shown after NHEK was treated with a series of dose of TCE 4 h later, and typical morphologic changes of apoptosis were also observed by TEM examination. FCM analysis revealed a sub-G(1) peak in the apoptotic cells. The apoptotic rate in TCE 0.125, 0.500, 2.000 mmol/L exposed groups (31.83%, 38.63%, 44.35%, respectively) were significantly higher than that in blank control (18.42%), while PI in TCE 0.125, 0.500, 2.000 mmol/L group (3.26%, 2.48%, 2.07%, respectively) were significantly lower than that in blank control (4.99%).

CONCLUSIONTCE may induce apoptosis of cultured NHEK in vitro, and inhibit cell proliferation through lipid peroxidation and oxidative stress.

Apoptosis ; drug effects ; Cells, Cultured ; Epidermis ; cytology ; drug effects ; Humans ; Keratinocytes ; drug effects ; Lipid Peroxidation ; Oxidative Stress ; Trichloroethylene ; toxicity

OBJECTIVEHeavy metals such as cadmium (Cd) are widely distributed in the environment as industrial pollutants and characterized by their ability to affect the male reproductive system. The objective of the present study was to test the effect of Cd in the concentration range from 10 to 1000 micromol/L, in vitro, on the membrane and DNA integrity, motility, and ability of sperm to undergo acrosomal exocytosis in Holstein bull spermatozoa.

METHODSBull semen samples were processed for sperm analyses using semen-diluting fluid, PBS. Membrane integrity of the processed bull sperm was evaluated by lipoperoxidation (LPO) test. Gelatin digestion test was performed to determine the ability of bull spermatozoa to undergo acrosomal exocytosis. Single cell gel electrophoresis (SCGE) assay was performed to detect the DNA strand breaks and alkali labile damages in the individual cell.

RESULTSWe found a significant increase in the lipoperoxidation (LPO) indicating the deleterious effect of Cd on the sperm membrane integrity. This effect was prominent at the concentration of 1000 micromol/L Cd. There was a negative correlation between LPO rate and the percentage of motile spermatozoa (r = -0.94, P < 0.001). The gelatin digestion test indicated that Cd caused a decline in the percentage of acrosomal exocytosis of bull spermatozoa. A reverse correlation was also found between LPO rate and the percentage of halos (r = -0.97, P < 0.001). Data obtained from the comet assay revealed that Cd was capable of inducing DNA breaks in the sperm nuclei. Almost 93% of DNA damages were double-stranded breaks. The correlation between LPO rate and the percentage of DNA breaks was found to be 0.95 (P < 0.001).

CONCLUSIONCollectively, Cd induced membrane impairments, lowered motility, DNA breaks and a decreased rate in the acrosome reaction of bull spermatozoa, leading to sperm dysfunction. Entering Cd in the male gonads and seminal plasma may exert deleterious effects on the animal sperm cells.

Acrosome Reaction ; Animals ; Cadmium ; toxicity ; Cattle ; DNA Breaks ; Lipid Peroxidation ; Male ; Semen ; drug effects ; Sperm Motility

OBJECTIVETo investigate the effect of carbon disulfide (CS(2)) on oxidation-antioxidation function of rat nerve tissues.

METHODSThirty male Wistar rats were randomly divided into the control group, the low-dosage exposure group and the high-dosage group, 10 rats each. The rats of the two exposure groups were administered with CS(2) by gavage at a dosage of 300 or 500 mgxkg(-1)xd(-1), 5 times every week for continuous 12 weeks. The alterations in glutathione (GSH), malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), hydrogen peroxidase (CAT) and total anti-oxidation (T-AOC) in cerebrum, spinal cord, and sciatic nerve of CS(2)-treated animals were assayed.

RESULTSThe results showed that the contents of MDA and ROS in nerve tissues of CS(2)-treated groups increased significantly except ROS in spinal cord and sciatic nerve of low dose group. The content of MDA was increased by 20.7% and 33.6% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 18.5% and 23.3% respectively in the spinal cord, and by 20.7% and 53.0% respectively in the sciatic nerve, The content of MOS was increased by 20.1% and 34.9% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, and by 14.1% and 15.4% respectively in the spinal cord and the sciatic nerve of the rats of the high-dosage group (P < 0.05 or P < 0.01). Furthermore, the activities of SOD, GSH-Px, CAT and T-AOC decreased significantly except GSH-Px and SOD in cerebrum of low dose group. The content of GSH was decreased by 17.2% and 26.5% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 26.4% and 31.2% respectively in the spinal cord, and by 15.1% and 20.0% respectively in the sciatic nerve. The content of T-AOC was decreased by 11.1 and 26.4% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 15.1% and 38.4% respectively in the spinal cord, and by 35.6% and 42.3% respectively in the sciatic nerve. The activity of SOD was decreased by 12.1% and 25.4% respectively in the spinal cord of the rats of the low-dosage group and the high-dosage group and by 16.4% and 30.3% respectively in the sciatic nerve. The activity of GSH-Px was decreased by 17.3% and 32.5% respectively in the spinal cord of the rats of the low-dosage group and the high-dosage group and by 17.1% and 21.5% respectively in the sciatic nerve. The activity of GSH-Px and SOD was decreased by 12.6% and 30.1% respectively in the cerebrum of the rats of the high-dosage group. The activity of CAT was decreased by 17.5% and 39.4% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 25.2% and 31.3% respectively in the spinal cord, and by 17.1% and 36.9% respectively in the sciatic nerve (P < 0.05 or P < 0.01).

CONCLUSIONSubchronic exposure to CS(2) can induce significant changes of oxidation-antioxidation function in rat nerve tissues, which might be related to CS(2)-induced neurotoxicity.

Animals ; Antioxidants ; metabolism ; Carbon Disulfide ; Lipid Peroxidation ; drug effects ; Nerve Tissue ; metabolism ; Rats ; Rats, Wistar

Adult ; Comet Assay ; DNA Damage ; drug effects ; Female ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Peracetic Acid ; toxicity

Homocysteine ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Lipid Peroxidation ; drug effects ; Metallothionein ; pharmacology

OBJECTIVETo investigate the chemical constituents of tuber of Gymnadenia conopsea.

METHODThe constituents were isolated by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and C-18, as well as reversed-phase HPLC. Structures of the isolates were identified by spectroscopic data analysis.

RESULTThirty-four compounds were isolated. Their structures were identified as six 2-isobutyltartrate benzyl ester glucosides: coelovirin A (1), coelovirin B (2), coelovirin E (3), coelovirin D (4), dactylorhin B (5) and loroglossin (6). Three 2-isobutylmalate benzyl ester glucosides: dactylorhin A (7), dactylorhin E (8) and militarine (9). Three lignans: arctigenin (10), lappaol A (11) and lappaol F (12). Six aromatic acid (alhyde or alcohol) derivatives: 4-beta-D-glucopyranosyloxyl-trans-phenylpropenoic acid (13), 4-beta-D-glucopyranosyloxyl-cis-phenylpropenoic acid (14), gastrodin (15), 4-beta-D-glucopyranosyloxylphenylaldehyde (16), 4-beta-D-glucopyranosyloxylbenzyl methyl ether (17), 4-beta-D-glucopyranosyloxyloxylbenzyl ethyl ether (18), and bis(4-hydroxybenzyl) ether mono 4-O-beta-D-glucopyranoside (19). Four cyclodipeptides: cyclo(L-Leu-L-Tyr) (20), cyclo(L-Leu-L-Pro) (21), cyclo(L-Val-L-Tyr) (22), and cyclo(L-Ala-D-Phe) (23). One N6-substituted andenosine: N6-(4-hydroxybenzyl)-adenine riboside (24). An aromatic amide: N-trans-feruloyltyramine (25). Nine aromatic acids (or aldehyde or alcohol): 3-hydroxybenzoic acid (26), 4-hydroxyisophthalic acid (27), 4-hydroxybenzyl alcohol (28), 4-hydroxybenzyl methyl ether (29), 4-hydroxybenzylaldehyde (30), 4-hydroxybenzoic acic (31), 4-hydroxy-3-methoxybenzoic acid (32), trans-p-hydroxyphenylpropenoic acid (33), and cis-p-hydroxyphenylpropenoic acid (34). At a concentration of 1.0 x 10(-6) mol x L(-1), compounds 10-12 showed antioxidative activity inhibiting Fe(+2) -cystine induced rat liver microsomal lipid peroxidation with inhibitory rates of 53%, 59%, and 52%, respectively(positive control VE with 35% inhibition).

CONCLUSIONThese compounds were obtained from the genus Gymnadenia for the first time except for 5-7, 9, 15, 28-34. Compounds 10-12 possess antioxidant activity.

Animals ; Lipid Peroxidation ; drug effects ; Microsomes, Liver ; drug effects ; metabolism ; Orchidaceae ; chemistry ; Plant Extracts ; analysis ; isolation & purification ; pharmacology ; Rats

Animals ; Biogenic Monoamines ; metabolism ; Brain ; drug effects ; metabolism ; Female ; Lipid Peroxidation ; drug effects ; Male ; Mice ; Paraquat ; toxicity

Adult ; Electrolysis ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Middle Aged ; Nickel ; adverse effects ; blood ; pharmacokinetics ; Occupational Exposure ; adverse effects

AIMTo study the relativity of the renocortical lipid peroxidation with renal tubules structure damage in renal injury induced by cisplatin in rats.

METHODSFemale Wistar rats were randomly divided into NS group, CDDP(I) group, CDDP(II) group and CDDP(III) group. All rats were injected via the tail vein with NS or cisplatin and NS qd in five days. The changes in content of Scr, BUN and MDA, the activity of SOD and GSH-Px of the renal cortex were measured. Alkaline phosphatase of renal tubular epithelia was stained by histochemistry and the slices of renal cortex were observed.

RESULTSThe contents of Scr and BUN of CDDP groups were significantly higher than those of NS group (P < 0.01). The content of renocortical MDA was significantly higher than that of NS group (P < 0.05). The activities of renoconical SOD and GSH-Px were lower than those of NS group (P < 0.05). The content of MDA, activities of renocortical SOD and GSH-Px with the content of Scr and BUN were significantly correlative. Alkaline phosphatase of renal tubular epithelia cells was losed largely and renal tubular epithelia cells were denaturative and necrotic partly in sections.

CONCLUSIONThe damage of renal cortex was correlative with its lipid peroxidation. The injury of renal cortex became heavier with cisplatin dose increased.

Animals ; Cisplatin ; adverse effects ; Female ; Kidney Cortex ; drug effects ; physiopathology ; Kidney Tubules ; pathology ; Lipid Peroxidation ; Rats ; Rats, Wistar

OBJECTIVETo establish an oxidative stress model induced by cumene hydroperoxide (cHP) in testis and epididymis of rats in vivo, and to understand the peroxidative damage of oxidative stress in testis, epididymal sperm and its propensity to induce nuclear DNA damage during spermatogenesis and sperm maturation in vivo.

METHODSAn organic hydroperoxide, cHP, 70% aqueous, diluted by 0.9% NaCl, was employed as model prooxidant. Ninety-day-old male Wistar rats were divided into a control and three cHP groups, and were administered intraperitoneally 0, 1/10, 1/6 and 1/4 LD50 cHP per day respectively at a dose of 2 ml/kg, for 7 consecutive days and were observed for any toxic symptoms and mortality. Twenty-four hours after the last dose, rats were sacrificed and induction of oxidative stress was ascertained by monitoring the degree of lipid peroxidation expressed as nano molar of malondialdehyde (MDA) in testicular homogenate and epididymal sperm. Nuclear DNA damage in testes and epididymal sperms was determined by comet assay. Motility of caudal sperms was counted and the morphology of testes and epididymis was observed under light microscope.

RESULTSRats of cHP administered groups were less vigorous than those of the control, but there were not death of rats during treatment. 1/10 LD50 per day for 7 consecutive days resulted in only a marginal increase in testicular MDA levels. However, 1/6 and 1/ 4 LD50 per day for 7 days of cHP administered to adult rats induced marked oxidative stress in testis and epididymal sperms as evidenced by a marked increase in MDA or nuclear DNA damage in testis and caput sperms, as well as significant decreases both in the body weight-and motility of caudal sperms. While the nuclear DNA damage caput sperms of 1/6 and 1/4 LD50 cHP administered rats increased significantly, nuclear DNA damage in caudal sperms showed no treatment related alterations.

CONCLUSIONOxidative stress in testis and epididymal sperms can be safely induced by applying multiple doses of cHP (1/6 and 1/4 LD50 per day for seven consecutive days). DNA damage caused by cHP induced oxidative stress may occurred mainly in testes.

Animals ; Benzene Derivatives ; toxicity ; DNA Damage ; Epididymis ; drug effects ; pathology ; Lipid Peroxidation ; drug effects ; Male ; Rats ; Rats, Wistar ; Sperm Count ; Spermatozoa ; drug effects ; pathology ; Testis ; drug effects ; pathology