OBJECTIVE: : To investigate the effect of chlorogenic acid (CGA) on non-enzymatic glycation and oxidation of low density lipoprotein (LDL). METHODS: : The non-enzymatic glycation incubation system of LDL-glucose was established. The contents of early glycation products (Amodori product) and intermediate products (dicarbonyl compound) were determined by ultraviolet-visible spectrophotometry, and the content of advanced glycation end products (AGEs) was determined by fluorescence spectrophotometry. The LDL oxidation incubation system was established. The contents of thiobarbituric acid reactive substances(TBARS) and conjugated diene were determined by ultraviolet-visible spectrophotometry. The tryptophan fluorescence quenching, and the content of lipofuscin, total fluorescence products, active aldehydes and malondialdehyde were determined by fluorescence spectrophotometry, and further verified by three-dimensional fluorescence spectroscopy. RESULTS: : In the LDL glycation experiment, 150 μg/mL and 300 μg/mL CGA inhibited the formation of Amadori product, dicarbonyl compounds and AGEs. In the LDL oxidation experiment, 15 μg/mL and 25 μg/mL CGA inhibited the formation of TBARS effectively; 5 μg/mL and 10 μg/mL CGA inhibited tryptophan fluorescence quenching, and the formation of active aldehydes, malondialdehyde, total fluorescence products, lipofuscin and conjugated diolefine. And the three-dimensional fluorescence spectroscopy showed the same results. CONCLUSIONS : CGA can inhibit non-enzymatic glycation and oxidation of LDL.
Chlorogenic Acid ; pharmacology ; Glycosylation ; drug effects ; Lipoproteins, LDL ; metabolism ; Oxidation-Reduction ; drug effects ; Thiobarbituric Acid Reactive Substances ; analysis

Animals ; Dust ; Male ; Malondialdehyde ; blood ; Mice ; Nanoparticles ; Oxidation-Reduction ; drug effects ; Particle Size ; Silicon Dioxide ; toxicity ; Superoxide Dismutase ; blood

AIMTo investigate the inclusion of coenzyme Q10 with beta-cyclodextrin (beta-CD).

METHODSThe inclusion of the electroactive guest molecule coenzyme Q10 with the host molecule beta-CD was studied by the polarography. The change of the reduction peak current of the inclusion complex with time and the change of the peak potential of the inclusion complex with beta-CD concentration were examined. In order to study the photostability, the change of the reduction peak current of both coenzyme Q10 and coenzyme Q10-beta-CD inclusion complex with time were also examined under light, separately.

RESULTSIn 0.1 mol x L(-1) HAc/NaAc (pH 4.7) buffer-ethanol/water (60:40) medium, coenzyme Q10 was included with p-CD to form an 1:1 inclusion complex. The formation constant Kf was 1.26 x 10(4) L x mol(-1) the apparent formation rate constant was 6.64 x 10(-2) min(-1). The photodegradation apparent rate constant of coenzyme Q10 as 7.77 x 10(-3) min(-1) and that of the coenzyme Q10-beta-CD inclusion complex was 3.38 x 10(-3) min(-1).

CONCLUSIONThe inclusion of coenzyme Q10 with beta-CD took place. The stability of coenzyme Q10 to lights was improved in a certain degree due to the formation of the inclusion complex.

Coenzymes ; chemistry ; Drug Compounding ; methods ; Light ; Oxidation-Reduction ; radiation effects ; Polarography ; methods ; Ubiquinone ; analogs & derivatives ; chemistry ; beta-Cyclodextrins ; chemistry

OBJECTIVETo investigate the effects of advanced oxidative protein products (AOPP) on the proliferation, apoptosis and matrix metalloproteinase-1 (MMP-1) synthesis of the human gingival fibroblast (HGF). To explore the possible mechanism of the periodontal destruction acceleration in diabetes through AOPP-mediated oxidative stress.

METHODSHGF were isolated by both tissue explant cultivation technique and enzyme digestion method. The culture media with 5, 50, 100 mg/L AOPP-HAS were added into each experimental group, but the culture media in the control group didn't contain AOPP-HAS. MTT colorimetric assay and ELISA were used to measure the changes of HGF proliferation and the levels of MMP-1 protein from HGF at different time periods, respectively. Seventy-two hours after co-culture with 50 mg/L AOPP-HSA, cell apoptosis was detected by flow cytometry with Annexin V/PI staining.

RESULTSCompared to the control group, the growth inhibition rate of HGF in 5, 50, 100 mg/L AOPP-HSA group was significantly different (P < 0.05). The peak value appeared at 48 hours of co-culture [(19.01 +/- 6.28)%, (30.48 +/- 5.75)%, (39.75 +/- 4.60)%, respectively]. There was a dose-dependent relationship between the growth inhibition rate and AOPP-HSA. No significant difference was detected on the apoptotic level between experimental group and the control (P > 0.05). The MMP-1 synthesis in 0.5, 5, 50, 100 mg/L AOPP-HAS group [(55.61 +/- 1.06), (65.78 +/- 4.04), (79.24 +/- 3.09), (89.76 +/- 28.88) mg/L, respectively] was significantly higher than that in the control [(34.90 +/- 3.15) mg/L] after 72 hours co-culture (P < 0.05). There was a dose-dependent relationship between MMP-1 and AOPP-HSA.

CONCLUSIONSAOPP may inhibit the proliferation of HGF and such effect was not achieved through apoptosis. AOPP may increase collagen degradation by promoting MMP-1 synthesis and thus may accelerate periodontal destruction process in diabetes.

Apoptosis ; drug effects ; Blood Proteins ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Gingiva ; cytology ; drug effects ; Humans ; Matrix Metalloproteinase 1 ; metabolism ; Oxidation-Reduction ; Oxidative Stress

OBJECTIVETo study the oxidative damage and apoptosis of renal tubular epithelial cells (NRK-52e cell line) induced by ethylbenzene.

METHODSNRK-52e cells were exposed to 30, 60, 90, 120 μmol/L ethylbenzene for 24 hours. Cell viability were measured using MTT, the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), the contents of malondialdehyde (MDA) and glutathione (GSH) were detected respectively. PI fluorescent staining assay was applied to detect percentage of apoptosis in ethylbenzene-treated groups.

RESULTSCompared with control group, cell outline became clear, cell diopter increased, cell became smaller and shrinkage, some cells broke in 60 μmol/L ethylbenzene-treated group. Plenty of cells died, suspension cells increased significantly in 90 μmol/L ethylbenzene-treated group. Compared with control group, cell viability the activities of SOD and CAT and the content of GSH were significantly decreased in 60 and 90 μmol/L ethylbenzene-treated groups (P<0.05). The MDA content were remarkably elevated in 90 μmol/L ethylbenzene-treated groups (P<0.05).

CONCLUSIONEthylbenzene can induce oxidative stress and apoptosis in NRK-52e cells (P<0.05).

Animals ; Apoptosis ; drug effects ; Benzene Derivatives ; toxicity ; Cell Line ; Epithelial Cells ; drug effects ; metabolism ; Kidney Tubules ; cytology ; Oxidation-Reduction ; Oxidative Stress ; drug effects ; Rats ; Reactive Oxygen Species ; metabolism

OBJECTIVETo investigate the effect of Coptis chinensis on oxidative hemolysis of erythrocytes in mice.

METHODAcetylphenyhydrazine (APH)-induced oxidative hemolysis of erythrocytes in mice were used. The contents of free hemoglobin of blood plasma, indirect bilirubin of serum, reticulocytes of blood, and malondialdehyde (MDA) of erythrocytes were measured. The activity of superoxide dismutase (SOD), reduced glutathione hormone (GSH), and glucose-6-phosp hate dehydrogenase (G-6-PD) of erythrocytes were also determined and the total-antioxygen capability (T-AOC) of blood was analyzed.

RESULTThe levels or amount of free hemoglobin of blood plasma, indirect bilirubin of serum, reticulocytes of blood and MDA of erythrocytes were higher in APH (0.03 g x kg(-1))-induced mice than normal mice. The activity or content of SOD, GSH and G-6-PD was lower in APH-induced mice than in normal mice. Primaquine (0.058 g x kg(-1)) could aggravated the degree of elevated hemolysis of erythrocytes in APH-induced mice. C. chinensis (0.6 g x kg(-1) could deprssed significantly the elevated levels of indirect bilirubin in serum. The levels of free hemoglobin of blood plasma, indirect bilirubin of serum, reticulocytes of blood, the production of SOD and GSH and T-AOC were also decressed by C. chinensis (0.6 g x kg(-1)).

CONCLUSIONC. chinensis suppressed t he degree of hemolysis of erythrocytes in APH-induced mice due to the suppression of the production of lipid peroxidation and increasing of the activity of antioxidase of erythrocytes.

Animals ; Coptis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Erythrocytes ; drug effects ; metabolism ; Female ; Hemolysis ; drug effects ; Male ; Mice ; Oxidation-Reduction ; Phenylhydrazines ; adverse effects ; Plasma ; metabolism ; Random Allocation

OBJECTIVETo study the mechanism of the reducing mechanism of hepatotoxicity induced by ethyl acetate fractions of Kansui Radix stir-baked with vinegar in mice.

METHODMice with normal ICR were orally administered with ethyl acetate fractions of Kansui Radix and Kansui Radix stir-baked with vinegar. Their blood and liver homogenate were collected to detect the level of AST, ALT, LDH, SOD, activities of Na(+) -K(+) -ATPase and Ca(2+) -Mg(2+) -ATPase, GSH and MDA. Liver tissues were collected for HE staining and morphological observation under light microscope.

RESULTAccording to the results of pathological sections, compared with the control group, all of Kansui groups showed a significant increase in the hepatic tissues injury (P < 0.01). Compared with Kansui groups, all of vinegar-baked groups showed a significant decrease in the hepatic tissues injury (P < 0.01). Compared with the control group, all of Kansui groups showed a significant increase in ALT, AST and LDH (P < 0.05, P < 0.001) in serum and hepatic tissues, and significantly decrease in the activity of SOD (P < 0.001) and the content of GSH. They also showed a significant increase in MDA (P < 0.001) and a significant decrease in the level of Na(+) -K(+) -ATPase and Ca(2+) -Mg(2+) -ATPase (P < 0.01) in hepatic tissues, with a certain dose-effect relationship. Compared with all of Kansui groups, all of vinegar-baked groups showed a significant decrease in ALT, AST and LDH (P < 0.05, P < 0.001), and a notable increase in SOD (P < 0.001) and GSH in serum and hepatic tissues. They also showed a remarkable decrease in MDA (P < 0.001), and a significant increase in the level of Na(+) -K(+) -ATPase and Ca(2+) - Mg(2+) -ATPase (P < 0.01) in hepatic tissues, with a certain dose-effect relationship.

CONCLUSIONBeing stir-baked with vinegar can significantly reduce the hepatotoxicity of Kansui Radix. Its mechanism may be related to the reduction of the effect of Kansui Radix on the permeability of hepatic tissues cell membranes and the oxidative injury.

Acetic Acid ; Animals ; Cell Membrane Permeability ; drug effects ; Euphorbia ; toxicity ; Liver ; drug effects ; metabolism ; pathology ; Mice ; Oxidation-Reduction ; Plant Extracts ; toxicity

Animals ; Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Cumulative Trauma Disorders ; metabolism ; Lipid Peroxidation ; drug effects ; Male ; Muscle, Skeletal ; injuries ; pathology ; Oxidation-Reduction ; Oxidative Stress ; drug effects ; Rats

OBJECTIVETo observe the effects of Modified Wenjing Decoction (MWJD) on the ovarian oxidative damage of cold coagulation blood stasis (CCBS) model rats, thus consummating the action mechanisms of MWJD for treating gynecopathy of CCBS syndrome.

METHODSFemale SD rats were randomly divided into the normal group, the model group, and the treatment group. Rats in the model group and the treatment group were placed in the ice water (0 -1 degrees C) to establish the rat model of CCBS syndrome. Meanwhile, MWJD was given by gastrogavage to rats in the treatment group for 2 weeks. Rats were decapitated during the oestrous cycle. The serum levels of estradiol (E2), progestone (P), testosterone (T) were detected. The bilateral ovarian tissues were taken out and operated on ice. The total bilirubin (TBIL), superoxide dismutase (SOD), malondialdehyde (MDA), total antioxygen capacity (T-AOC) were detected after homogenate.

RESULTSCompared with the normal group, serum levels of E2, P, T, and ovarian levels of TBIL, SOD, T-AOC all obviously decreased in the model group, and the ovarian MDA level increased, showing statistical difference (P <0. 05, P <0.01). After treated by MWJD, when compared with the model group, serum levels of E2, P, T, and ovarian levels of TBIL, SOD, T-AOC increased, the ovarian MDA level decreased, showing statistical difference (P <0.05, P <0.01). All indices except serum T level and the ovarian MDA content restored to the normal levels (all P>0.05).

CONCLUSIONSThe ovarian functions are abnormal in CCBS model rats. MWJD could restore the ovarian functions by improving the ovarian oxidative damage state, thus treating gynecopathy of CCBS syndrome.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Malondialdehyde ; metabolism ; Ovary ; drug effects ; metabolism ; pathology ; Oxidation-Reduction ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo establish the oxidative damage model of cochlea hair cells using organic oxidant t-BHP in vitro.

METHODSHEI-OC1 cells were exposed to t-BHP at 8 doses (30~4000 µmol/L) for 12 h. Trypan blue test was used to detected the cellular viability and MTT assay was utilized to measured the cellular proliferation. The intracellular ROS levels were determined by 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA).

RESULTSThe survival rates of HEI-OC1 cells started decrease significantly at the dose of 100 µmol/L t-BHP, the peak of decreased survival rates appeared at the doses of 200~800 µmol/L. The results of MTT assay demonstrated that 30 µmol/L t-BHP could promote cellular proliferation ability, when t-BHP concentrations were higher than 200 µmol/L, the cellular proliferation ability was inhibited. The results of DCFH-DA assay showed that there was no fluorescence in control group, the strong fluorescence was observed in positive control group, the weak fluorescence was observed in 30~50 µmol/L t-BHP groups, the bright fluorescence was observed in 100 µmol/L t-BHP group, still the stronger fluorescence was observed in 200~1000 µmol/L groups, but the cellular number decreased with the doses because of the lower cellular viability.

CONCLUSIONThe exposure to 100 µmol/L t-BHP for 12 h could simulate the oxidative damage induced by noise in cochlear hair cells.

Cell Survival ; Cells, Cultured ; Hair Cells, Auditory ; drug effects ; pathology ; Humans ; Noise ; adverse effects ; Oxidation-Reduction ; Oxidative Stress ; Reactive Oxygen Species ; analysis ; tert-Butylhydroperoxide ; toxicity