Apoplastic redox metabolism: Synergistic phenolic oxidation and a novel oxidative burst

The plant apoplast is an important mediator of communication between the cell cytoplasm and its surroundings. Plant cell suspensions offer a convenient model system to gain insight into apoplastic physiology. Here, we describe a novel phenomenon that took place when two naturally occurring phenolics were added together to either soybean or tobacco cell suspensions. Acetosyringone (AS) and/or hydroxyacetophenone (HAP), phenolics found in the extracellular/apoplast of tobacco cells, were added to soybean or tobacco cell suspensions undergoing an oxidative burst. Individually, AS appeared to be utilized as a typical peroxidase substrate to scavenge hydrogen peroxide, while HAP was utilized at a much lower rate. However, when added together the rate of utilization of both phenolics increased and surprisingly resulted in the production of hydrogen peroxide. We have further characterized this novel phenomenon in suspension cells. This study demonstrates that certain phenolics in plants can cause co-oxidation which, as in animals, could alter the structure and bioactivity of surrounding phenolics.     

低氧胁迫下黄瓜幼苗根系多胺代谢的变化

 利用外源Spd、PAO 抑制剂研究了黄瓜幼苗根系在低氧水培时的生长状况以及内源PAs、H₂O₂ 含量和PAO活性的动态变化。结果表明, 外源PAs对低氧胁迫下黄瓜幼苗根系的生长有促进作用;低氧处理后3种PAs含量都明显上升, 外源Spd和PAO抑制剂增加了根系中的PAs含量, 降低了根系中H₂O₂ 含量, 减缓了低氧伤害; PAs为低氧胁迫的应激产物, PAO为适应酶类, 其活性随PAs含量的变化而发生变化。     

Induction of apoptosis with Salvia miltiorrhiza monomer IH764-3 via downregulating focal adhesion kinase in H2O2-stimulated hepatic stellate cells

AIM: To investigate the effect of IH764-3 on the apoptosis of H₂O₂-stimulated hepatic stellate cells(HSCs) and the alteration of focal adhesion kinase (FAK). METHODS: The proliferation of HSCs was examined by direct cell count and the apoptosis was determined by Annexin-V/PI labeling, while the morphological change was observed by light microscopy and transmission electron microscopy. In addition, FAK mRNA was detected by RT-PCR. RESULTS: H₂O₂ promoted the proliferation of HSCs. IH764-3 induced the apoptosis of HSCs in a dose-dependent manner. The HSC apoptotic rates of different groups were 6. 35%,9. 28%,15. 10%,19. 69%,respectively, after treated with different concentrations of IH764-3 for 48 h while H₂O₂ group showed 2. 30%. In 30 mg/L group, the apoptosis rates were 6. 73%、10. 34%、15. 10% for the indicated time periods(12 h, 24 h, 48 h). In the presence of IH764-3, FAK mRNA decreased. The FAK mRNA reduction began at 2 h after adding IH764-3. CONCLUSION: IH764-3 induced the apoptosis of HSCs. Down-regulating the expression of FAK mRNA may be one of its mechanisms.     

Protective effect of HSP70 on injuried myocardial cells induced by H2O2

AIM: To investigate the role of heat-shock protein 70(HSP₇₀) in the protection of myocardial cells against ischemic injury.METHODS: Myocardial cells were cultured in vitro. HSP₇₀ was induced by hyperthermia. H₂O₂-injured myocardial cells were divided into different groups. Flow cytometry, DNA Ladder and biochemistry methods were employed to observe the myocardial cells of different groups.RESULTS: Immunohistochemistry showed hyperthermia induced the up-regulation of HSP₇₀ in myocardial cells. Apoptotic rate, activity analysis of cytochrome C and succinic dehydrogenase in H₂O₂-injuried and HSP₇₀-protected groups were obviously different. Electron micrograph shomed hyperthermia alliviated myocardial cell injury induced by H₂O₂. CONCLUSION: HSP₇₀ delays apoptosis and protects against H₂O₂-induced myocardial cell injury.     

Evaluation of chemical disinfestants in reducing Verticillium dahliae conidia in irrigation water

Irrigation water disinfestation is an unexplored option for reducing Verticillium dahliae inoculum in water and consequently for more efficiently managing Verticillium wilts in Andalusia. We assessed Suppressive Efficacy (SE; water was infested and subsequently treated) and Preventive Efficacy (PE; previously treated water was subsequently inoculated) of OX-VIRIN^®, OX-AGUA AL 25^® and Deccoklor^® in reducing water infestations by V. dahliae conidia. Five concentrations of each disinfestant, the lowest three being recommended by the manufacturer, were tested in vitro against six V. dahliae isolates. Validation assays were carried out in experiments under natural environmental conditions in spring. The four highest concentrations of OX-VIRIN^® (0.8–51.2 mL L⁻¹), the three highest of OX-AGUA AL 25^® (46.4–417.5 μL L⁻¹) and the two highest of Deccoklor^® (0.375 and 3.75 mL L⁻¹), showed an in vitro-efficacy (SE and PE) of 96.2, 80.0 and 100.0% after 30, 5 to 30 and 15 days respectively. Therefore, recommended concentrations for OX-VIRIN^® and OX-AGUA AL 25^® showed a greater in vitro-effectiveness than those recommended for Deccoklor^®. Assays in natural environmental conditions proved that OX-VIRIN^® at the recommended concentration of 3.2-mL L⁻¹, applied every 28 days to water, was the most effective treatment (SE and PE), with a 100% reduction of the average relative viability after 56 days. Other chemical treatments showing high in vitro-efficacy, such as OX-VIRIN^® at 0.8 mL L⁻¹ and OX-AGUA AL 25^® at 46.4 μL L⁻¹ showed an SE of 99.9% after 14 and 28 days when applied every 28 and 14 days, respectively. However, PE of OX-AGUA AL 25^® at 46.4 μL L⁻¹ was only 59 and 38% after 28 and 14 days respectively, depending on the experiment.     

对蜂蜜中主要抗褐变因子的探讨

选用ＡＧ５０Ｗ－Ｘ８阳离子交换树脂，并以批处理的方式，成功地将蜂蜜中的肽类物质与其它组合得到了分离。抗褐变实验结果清楚地表明，蜂蜜中的肽类成分完全没有抑制褐变的活性。这与以前的报道截然不同。     

Polyamines enhance chilling tolerance of cucumber (Cucumis sativus L.) through modulating antioxidative system

Cucumber chilling-resistant cultivar Changchun mici and -sensitive cultivar Beijing jietou were used in this study to investigate the effects of exogenous PAs on protection against chilling injury as well as on changes of physiological features, and the fluctuation of free PAs content in the leaves under chilling stress. Upon chilling treatment, free spermidine (Spd), spermine (Spm) and putrescine (Put) were remarkably induced in the leaves of cv. Changchun mici 1 day after treatment. The induction of Put declined thereafter, whereas Spd and Spm levels increased steadily. In the leaves of cv. Beijing jietou, Put content was increased only at 1 day after chilling while Spd content decreased significantly upon chilling treatment. Chilling reduced soluble protein content, and decreased the activities of antioxidant enzymes, including superoxidase dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) only in Beijing jietou. However, these changes could be renovated by exogenous application of Put and Spd. It was also found that pretreatment with Put and Spd diminished the increased electrolyte leakage and malondialdehyde (MDA) content caused by chilling in the leaves of both cultivars. Pretreatment of methyglyoxal-bis-(guanylhydrazone) (MGBG), the PAs biosynthetic inhibitor cancelled the effects of PAs in most of the treatments. Moreover, histochemical staining and quantitative measurements showed that exogenous application of Put and Spd eliminated but MGBG exaggerated the hydrogen peroxide (H₂O₂) accumulation caused by chilling stress, especially in leaves of Beijing jietou. Interestingly, Changchun mici was found to contain higher endogenous free PAs contents compared to Beijing jietou. While no significant difference of SOD, POD and CAT activities was found between non-chilling Changchun mici and Beijing jietou seedlings, the former exhibited higher APX activity than the latter. These results suggest that PAs play important roles in the tolerance of cucumber against chilling stress, which is most likely achieved by acting as oxidative machinery against chilling injury.     

Effect of exogenous hydrogen sulfide on lipophagy in mouse primary hepatocytes

AIM: To evaluate the effect of exogenous hydrogen sulfide (H₂S) from GYY4137 on lipophagy in mouse primary hepatocytes. METHODS: The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion method were divided into 4 groups: the cells in control group were incubated with normal medium; the cells in model group were incubated with 1.2 mmol/L oleic acid (OA) for 48 h; the cells in H₂S group or propargylglycine (PAG) group were incubated with 1.2 mmol/L OA for 48 h followed by serum-free phenol red-free RPMI-1640 medium which contained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h. The cells were collected to conduct immunofluorescence staining of LC3 and photography under fluorescence microscope, phase-contrast microscope or transmission electron microscope. The protein expression of LC3-Ⅰ/Ⅱ in the hepatocytes was determined by Western blot. RESULTS: In contrast with the model group, the fluorescent particles of LC3, the protein expression of LC3, the number of autophagic lysosome and vacuoles in hepatocytes in H₂S group increased. CONCLUSION: In steatosis hepatocytes, exogenous H₂S promotes the lipophagy.     

Effect of GYY4137 on cytosolic lipid decomposition in mouse primary hepatocytes

AIM: To evaluate the effect of GYY4137, a novel hydrogen sulfide (H₂S) donor, on cytosolic lipid decomposition in mouse primary steatosis hepatocytes. METHODS: Oleic acid (OA) was used to induce hepatic steatosis model in vitro. The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion were divided into 4 groups:the cells in control group were incubated with normal medium for 54 h; the cells in model group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 for 6 h; the cells in H₂S group or DL-propar-gylglycine (PAG; an inhibitor of cystathione γ-lysase, inhibiting H₂S synthesis) group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 which contained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h. The glycerin release and the protein expression of hormone-sensitive lipase (HSL) in the cells were mea-sured. RESULTS: Compared with model group, the glycerin release and the protein expression of phosphorylated HSL (p-HSL) in H₂S group decreased significantly, while those increased significantly in PAG group. CONCLUSION: In steatosis hepatocytes, exogenous H₂S possibly decreases cytosolic lipid decomposition by decreasing the protein level of p-HSL.