扶正解毒汤对染镍体外培养人支气管上皮细胞抗氧化酶活性及丙二醛含量的影响

目的:研究灌服中药扶正解毒汤(FJD)后兔血清拮抗硫酸镍(NiSO4)的细胞毒作用及其机制。方法:以NiSO4和不同剂量FJD含药血清共同处理体外培养人支气管上皮(16HBE)细胞,测定其存活率及活力,同时观察细胞内谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性以及丙二醛(MDA)含量的变化。结果:在镍暴露细胞培养体系中加入FJD含药血清,可提高该细胞的存活率和活力,增强细胞内GSH-Px及SOD活性,减少MDA的生成,其中以高剂量FJD含药血清的作用最为明显。结论:NiSO4可诱发细胞氧化应激的产生,FJD具有拮抗硫酸镍细胞毒性的作用,可能与其抗氧化应激功能有关。

Physiological characteristics changes of Aesculus chinensis seeds during natural dehydration

A study was conducted to determine the physiological characteristics changes of Aesculus chinensis seeds during natural dehydration in 2003. The results showed that A. chinensis seeds were recalcitrant with being highly desiccation-sensitive. The seed moisture content of fresh fruits was higher than 60%. When the seeds were naturally dried for 30 days, their moisture content declined to 30.2% and their viability was completely lost. The seed germination percentage had a small increase at the beginning of desiccation and then decreased rapidly. The relative electrical conductivity of the A. chinensis seeds increased along with a decrease in seed moisture content. However, there was an abnormal increase in relative electrical conductivity when the seed moisture content was between 53.7% and 50.9%. Superoxide dismutase （SOD） activity decreased rapidly in the period of desiccation except for an abnormality when the seed moisture content was between 53.7% and 50.9%. Malondialdehyde （MDA） content increased slowly at the early stage of desiccation and then rose rapidly after the moisture content was below 50.9%. The soluble sugar content in seeds slowly increased with the increasing period of desiccation. The seed germination percentage was at the high level when seed moisture content was in range of 47%- 60%, which suggests that this was the optimum moisture content for maintaining A. chinensis seed viability.

Occurrences of Yttrium in Soil and Its Potential Impacts on Paddy Rice Triticum aestivum

[Objective] The effects of yttrium nitrate （YNO3） on biomass and antioxi- dant systems of paddy rice （Yttrium （Y）; Oxidative stress; Dismutases （SOD）; Per- oxidases （POD）, Catalases （CAT）, Paddy rice （Triticum aestivum）） together with the occurrences of Y in soils were investigated to assess its ecotoxicological effects on plant. [Method]Y solutions with various concentrations were sprinkled on soil sam- ples, which were well mixed and then put into culture dishes to culture paddy rice seeds for further evaluation. [Result] The results indicated that 25-100 mg/kg Y treatments significantly increased the biomass （total weight, root weight, shoot weight and leaf weight）, chlorophyll （CHL） content and protein content of paddy rice, whereas 200-800 mg/kg Y treatments had a converse effect. Similarly, biomarker for the antioxidant systems including superoxide dismutases （SOD）, peroxidases （POD） and catalases （CAT） all exhibited similar trends in both shoots and roots of paddy rice. At the same time, the malonaldehyde （MDA） content increased at from 25 to 100 mg/kg and decreased with concentrations of Y from 100 to 800 mg/kg in both shoots and roots of paddy rice. This indicated that Y could stimulate the growth of plant at low concentration, but inhibit the growth at relatively high concen- tration. [Conclusion] The levels of Y were 641＋49, 328_＋16 and 473_＋40 mg/kg in soils collected from mining area, farmland and navel orange orchard respectively. The levels of Y in the investigated area were higher than the benefit level （100 mg/kg）, which could cause low biomass as well as low activity of SOD, POD and CAT in paddy rice. Therefore, a more careful use of Y is necessary in crop management.

Mechanism of Oxidative Burst in Tobacco Leaves and Cells Induced by Palmin from Phytophthora palmi

In order to reveal the signaling pathways triggered by elicitor in plant-microbe interactions, the mechanisms of hypersensitive necrosis responses in Nicotiana tabacum L. cv. Gexin III induced by palmin were studied at molecular and cellular level. The burst of superoxide, intercellular diffusion of hydrogen peroxide and process of cell death induced by palmin were investigated in tobacco plants by biochemical methods and Confocal microscopy. The results showed that a large amount of O-2(.-) was rapidly generated in tobacco cell elicited by palmin as a result of activation of NADPH oxidase, and the O-2(.-) was dismutated into H2O2 immediately by superoxide dismutase (SOD). Accumulation and intercellular diffusion of H2O2 were shown to be a trigger for hypersensitive cell death; and Ca2+ and some specific protein kinase were also shown to be involved in the activation of oxidative burst in tobacco cell induced by palmin.

Enzyme responses and lipid peroxidation in gills and hepatopancreas of clam Mactra vereformis, following cadmium exposure

To assess the toxicity of heavy metal pollution to marine intertidal shellfish, enzymatic responses and lipid peroxidation were investigated in the clam Mactra vereformis exposed to cadmium under laboratory conditions. Three antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPx), two immune defense enzymes (acid phosphatase, ACP; alkaline phosphatase, ALP), and one lipid peroxidation product (malondialdehyde, MDA) were measured in the gills and the hepatopancreas of the clam exposed to 0, 25, 75, and 125 μg/L cadmium for 0, 1, 3, 5, and 7 d. The results show that the concentrations of antioxidant enzymes in the organs soared to a peak value on the first day and then decreased afterwards in most cases. CAT and GPx activities in the hepatopancreas were higher than in the gills, but the SOD activity was lower in the hepatopancreas. ACP activity was unchanged until Day 3 in the hepatopancreas and until Day 5 in gills, when it began to increase. ALP activity showed no significant relationship with Cd treatment. MDA concentrations increased in the two tissues after Cd exposure, peaked on Day 3 in gills, and on Day 5 in hepatopancreas. These observations show that changes in the activities of antioxidant enzymes and ACP reflect the time course of oxidative stress in the clam caused by Cd, and could be used as potential biomarkers for ecotoxicological bioassays of heavy metals.

Effects of UV-B irradiation on isoforms of antioxidant enzymes and their activities in red alga Grateloupia filicina(Rhodophyta)

Macroalgae in a littoral zone are inevitably exposed to UV-B irradiance. We analyzed the effects of UV-B on isoenzyme patterns and activities of superoxide dismutase （SOD）, peroxidase （POX）, catalase （CAT）, and ascorbate peroxidase （APX） of red algae Grateloupia filicina （Lamour.） C. Agardh. The activities of SOD, CAT, and APX changed in response to UV-B in a time- and dose-dependent manner. POX activity increased significantly under all three UV-B treatments. The enzymatic assay showed three distinct bands of SODI （Mn-SOD）, SODII （Fe-SOD）, and SODIII （CuZn-SOD） under a low （Luv） and medium （Muv） dose of UV-B irradiation, while SODI and SODIII activities decreased significantly when exposed to a high dose of UV-B irradiation （Huv）. The activity of POX isoenzymes increased significantly after exposure to UV-B, which is consistent with the total activity. In addition, a clear decrease in activity of CATIV was detected in response to all the three doses of UV treatments. Some bands ofAPX isoenzyme were also clearly influenced by UV-B irradiation. Correspondingly, the daily growth rate declined under all the three exposure doses, and was especially significant under Muv and Huv treatments. These data suggest that, although the protection mechanisms of antioxidant defense system are partly inducible by UV-B to prevent the damage, G.filicina has incomplete tolerance to higher UV-B irradiation stress.

Use of antioxidant enzymes of clam Ruditapesphilippinarum as biomarker to polycyclic aromatic hydrocarbon pollution

The typical organic pollutant polycyclic aromatic hydrocarbon(PAH) anthracene was selected as a contaminant to investigate its eff ects on the activities of superoxide dismutase(SOD), catalase(CAT) and glutathione peroxidase(GSH-Px) in the clam Ruditapes philippinarum. The results show that SOD, CAT and GSH-Px had diff erent induction and inhibition reactions to anthracene stress, and that three diff erent organs in R. philippinarum(visceral mass, muscle tissue and mantle) had diff erent sensitivities to anthracene stress. This study suggest that SOD activities of the visceral mass, CAT activitities of the mantle and the visceral mass, and GSH-Px activity of the muscle tissue could be used as sensitive indicators of anthracene stress in R. philippinarum.

Effect of Nitric Oxide on Esophageal Cancer Cell Line TE-1

Objective To investigate the radiosensitizing effect of nitric oxide(NO) combined with radiation on esophageal cancer cell line TE-1.Methods Methyl thiazolyl tetrazolium(MTT) assay was used to assess the effects of NO and radiation on TE-1 cells regarding inhibition of cell proliferation.Flow cytometry was used to examine the effect of NO and radiation on cell apoptosis and cycle.Reverse transcription polymerase chine reaction and Western blot were used to evaluete the effect of NO on mRNA and protein expression of manganese superoxide dismutase(MnSOD).Results NO inhibited the proliferation of TE-1 cells while significantly enhancing their radiosensitivity.The application of NO combined with radiation significantly increased the apoptosis rate and G2/M phase proportion of TE-1 cells,with substantial decreases in the MnSOD mRNA and protein expression levels.Conclusions NO reduces the MnSOD mRNA and protein expression levels by affecting TE-1 cell cycle,further inhibiting the apoptosis of esophageal cancer cells and enhancing the killing effect of radiation on esophageal cancer cells.

Purification and characterization of iron-cofactored superoxide dismutase from Enteromorpha linza

A superoxide dismutase was purified from Enteromorpha linza using a simple and safe procedure, which comprised phosphate buffer extraction, ammonium sulphate precipitation, ion exchange chromatography on Q-sepharose column, and gel filtration chromatography on Superdex 200 10/300GL. The E. linza superoxide dismutase （E/SOD） was purified 103.6-fold, and a yield of 19.1% and a specific activity of 1 750 U/rag protein were obtained. The SDS-PAGE exhibited E/SOD a single band near 23 kDa and the gel filtration study showed E/SOD＇s molecular weight is near 46 kDa in nondenatured condition, indicating it＇s a homodimeric protein. E/SOD is an iron-cofactored superoxide dismutase （Fe-SOD） because it was inhibited by hydrogen peroxide, insensitive to potassium cyanide. The optimal temperature for its maximal enzyme activity was 35℃, and it still had 29.8% relative activity at 0℃, then E/SOD can be classified as a cold-adapted enzyme. E/SOD was stable when temperature was below 40℃ or the pH was within the range of 5 10. The first 11 N-terminal amino acids orE/SOD were ALELKAPPYEL, comparison of its N-terminal sequence with other Fe-SOD N-terminal sequences at the same position suggests it is possibly a chloroplastic Fe-SOD.

Keratinocyte growth factor gene therapy ameliorates ulcerative colitis in rats

AIM:To investigate the effect of keratinocyte growth factor(KGF) gene therapy in acetic acid-induced ulcerative colitis in rat model.METHODS:The colitis of Sprague-Dawley rats was induced by intrarectal infusion of 1 mL 5%(v/v) acetic acid.Twenty-four hours after exposed to acetic acid,rats were divided into three experimental groups:control group,attenuated Salmonella typhimurium Ty21a strain(SP) group and SP strain carrying human KGF gene(SPK) group,and they were separately administered orally with 10% NaHCO3,SP or SPK.Animals were sacrificed and colonic tissues were harvested respectively on day 3,5,7 and 10 after administration.Weights of rats,colonic weight/length ratio and stool score were evaluated.Histological changes of colonic tissues were examined by hematoxylin and eosin(HE) staining method.The expression of KGF,KGF receptor(KGFR) and TNF-α were measured either by enzyme-linked immunosorbent assay or Western blotting.Immunohistochemistry was used to detect the cellular localization of KGFR and Ki67.In addition,superoxide dismutase(SOD) activity and malondialdehyde(MDA) contents in the homogenate were measured.RESULTS:Body weight and colonic weight/length ratio were declined in SPK group compared with SP and control groups(body weight:272.78 ± 17.92 g vs 243.72 ± 14.02 g and 240.68 ± 12.63 g,P < 0.01;colonic weight/length ratio:115.76 ± 7.47 vs 150.32 ± 5.99 and 153.67 ± 5.50 mg/cm,P < 0.01).Moreover,pathological changes of damaged colon were improved in SPK group as well.After administration of SPK strain,KGF expression increased markedly from the 3rd d,and remained at a high level till the 10th d.Furthermore,KGFR expression and Ki67 expression elevated,whereas TNF-α expression was inhibited in SPK group.In the group administered with SPK,SOD activity increased significantly(d 5:26.18 ± 5.84 vs 18.12 ± 3.30 and 18.79 ± 4.74 U/mg,P < 0.01;d 7:35.48 ± 3.35 vs 22.57 ± 3.44 and 21.69 ± 3.94 U/mg,P < 0.01;d 10:46.10 ± 6.23 vs 25.35 ± 4.76 and 27.82 ± 6.42 U/mg,P < 0.01) and MDA contents decreased accordingly(d 7:7.40 ± 0.88 vs 9.81 ± 1.21 and 10.45 ± 1.40 nmol/mg,P < 0.01;d 10:4.36 ± 0.62 vs 8.41 ± 0.92 and 8.71 ± 1.27 nmol/mg,P < 0.01),compared with SP and control groups.CONCLUSION:KGF gene therapy mediated by attenuated Salmonella ameliorates ulcerative colitis induced by acetic acids,and it may be a safe and effective treatment for ulcerative colitis.

Enhancement of Superoxide Dismutase and Catalase Activities and Salt Tolerance of Euhalophyte Suaeda salsa L. by Mycorrhizal Fungus Glomus mosseae

Arbuscular mycorrhizal （AM）-mediated plant physiological activities could contribute to plant salt tolerance. However, the biochemical mechanism by which AM fungi enhance salt tolerance of halophytie plants is unclear. A pot experiment was conducted to determine whether salt tolerance of the C3 halophyte Suaeda salsa was enhanced by the AM fungus Glomus rnosseae. When 60-day-old S. salsa seedlings were subjected to 400 mmol L-1 NaC1 stress for 35 days, plant height, number of leaves and branches, shoot and root biomass, and root length of G. mosseae-colonized seedlings were significantly greater than those of the nonmycorrizal seedlings. Leaf superoxide dismutase （SOD） activity at all sampling times （weekly for 35 days after salt stress was initiated） and leaf catalase （CAT） activity at 2 and 3 weeks after salt stress was initiated were also significantly enhanced in G. mosseae-colonized S. salsa seedlings, while the content of leaf malondialdehyde （MDA）, a product of membrane lipid peroxidation, was significantly reduced, indicating an alleviation of oxidative damage. The corresponding leaf isoenzymes of SOD （Fe-SOD, Cu/Zn-SOD1, and Cu/Zn-SOD2） and CAT （CAT1 and CAT2） were also significantly increased in the mycorrhizal seedlings after 14 days of 400 mmol L-1 NaC1 stress. Our results suggested that G. rnosseae increased salt tolerance by increasing SOD and CAT activities and forming SOD and CAT isoforms in S. salsa seedlings.

Research on Fuzi based on animal thermotropism behavior to discover if it has fewer "hot" characteristics without Ganjiang

OBJECTIVE:To investigate whether Fuzi(Radix Aconiti Praeparata) has fewer "hot" characteristics when administered without Ganjiang(Rhizoma Zingiberis).METHODS:Differences in the thermotropism behaviors of mice treated either with Fuzi(Radix Aconiti Praeparata),Ganjiang(Rhizoma Zingiberis) or the combination of the two given intragastrically were investigated using the Animal Thermotropism Behavior Surveillance System.The water intake volume,oxygen consumption volume,adenosine triphosphatase(ATPase) activity,total antioxidant capacity(T-AOC) and total superoxide dismutase(T-SOD) activity were determined during the investigation.RESULTS:When Fuzi and Ganjiang were administered together,the rate at which mice remained on a warm plate("remaining rate") and the times and distances of their movement were all significantly reduced(P<0.05).Compared with the Normal group,the reduction was 55.1%,48.3% and 44.8%,while compared with the Fuzi group,the reduction was 57.6%,34.3% and 36.0%,indicating that "cold" tropism was significantly increased.Compared with the Normal and Fuzi groups,the ATPase activity and the respiratory oxygen consumption volume of the Fuzi + Ganjiang group were significantly increased(P<0.05),suggesting an improvement in energy metabolism and showing a "hot" characteristic when Fuzi and Ganjiang are present together.Additionally,the T-AOC and T-SOD activity were significantly enhanced(P<0.05).CONCLUSION:The behavior of mice tending toward "cold" tropism can be regarded as a quantitative reflection of Fuzi having fewer characteristics consistent w ith a "hot" nature when not used with Ganjiang,the functional mechanism of which may be a change in the ATPase activity in liver tissue.

Influences of electro-acupuncture at related Jing-well Points in rats with vascular dementia

OBJECTIVE:To observe the effects of electro-acupuncture(EA) at related Jing-well Points(HT 9,PC 9,KI 1 and LU 11) in rats with vascular dementia(VD) and discuss the relative mechanism.METHODS:A randomized controlled animal experiment was designed.A total of 104 rats were involved in the present study and divided randomly into 4 groups:sham-operation group,model group,Jing-well Points group,and medication group.The VD model was established according to the modified 4-vessel occlusion(4-VO) method.VD rats in the Jing-well Points group were treated by EA at the related Jing-well Points(HT 9,PC 9,KI 1 and LU 11) while those in the medication group were treated with nimotop.The step-down avoidance test was performed before and after treatment in all rats.Latency and error frequency indexed memory function were recorded.Nitric oxide(NO) levels and superoxide dismutase(SOD) activity in both cerebral tissue and serum were detected after the treatment course.RESULTS:A total of 42 rats were included in the final analysis.Compared with the model group,the latency in the Jing-well Points group was significantly prolonged(P<0.01) and the error frequency was significantly decreased(P<0.05) after therapy;the decrease in NO levels in both brain tissue and serum was significant(P<0.05 and P<0.01,respectively);and the increase in SOD activity was also significant(P<0.01).There was no significant difference in latency,error frequency,NO levels and SOD activity between the Jing-well Points group and medication group.CONCLUSION:EA at related Jing-well Points can remarkably improve memory impairment in VD rats.Moreover,decreasing the overproduction of NO and strengthening the ability of eliminating free radicals may provide therapeutic potential for the treatment of VD.