与NADPH氧化酶相关的植物抗病性研究进展

综述了烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶及其催化产生的活性氧在植物抗病过程中的作用。     

Accumulation of H<Subscript>2</Subscript>O<Subscript>2</Subscript> in xylem fluids of cucumber stems during ASM-induced systemic acquired resistance (SAR) involves increased LOX activity and transient accumulation of shikimic acid

Systemic acquired resistance (SAR) to Colletotrichum orbiculare was induced in young cucumber (Cucumis sativus) plants within 3 h of ASM (acibenzolar-S-methyl) application onto the first leaves. A potent signal associated with significant accumulation of hydrogen peroxide in xylem fluids from severed stems appeared to be rapidly translocated from elicited lower leaves within 3 h and 6 h after treatment. Some metabolites of the shikimate, phenylpropanoid and lignin biosynthetic pathways were quantified and significant increases in the levels of shikimic acid were observed in ASM-treated plants challenge-inoculated with the anthracnose fungus. Furthermore, the expression of the 5-enolpyruvylshikimate-3-phosphate synthase gene (EPSPS) was 1.5 times higher within 12 h after ASM treatment in challenge-inoculated plants than in the untreated control. The involvement of lipoxygenase activity, shikimic acid and others such as caffeic acid in the induction of SAR is discussed.     

Characterization and expression of NADPH oxidase genes in stimulated splenic neutrophils of tilapia,Oreochromis niloticus by a mix of traditional Chinese medicine and Bacillus species (TCMBS)

Traditional Chinese medicine and Bacillus species (TCMBS) mixture is an immunostimulant with considerable promise as an alternative in improving fish health. However, nothing is known on its effects on the expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase genes and production of reactive oxygen species (ROS) in the neutrophils of fish. The full lengths of tilapia phagocytic NADPH oxidase genes gp91^phox, p22^phox, p40^phox, p47^phox, and p67^phox were cloned and their expression profiles after TCMBS stimulus investigated. The cDNAs of tilapia gp91^phox, p22^phox, p40^phox, p47^phox, and p67^phox contained open reading frames of 1698 bp, 561 bp, 1053 bp, 1584 bp, and 1470 bp respectively, encoding 561, 186, 350, 527, and 489 amino acids respectively. Comparison of the deduced amino acid sequences showed that tilapia NADPH oxidase genes shared 58%–91% and 21%–67% identity with those of other teleost and mammals respectively. Besides, tilapia NADPH oxidase genes contain conserved domains and motifs required for ROS generation. Phylogenetic analysis suggested tilapia NADPH oxidase genes were close to those of Fundulus heteroclitus. After 2 weeks of TCMBS application showed significant upregulation in expression of NADPH oxidase genes, antioxidant genes (i.e., superoxide dismutase, catalase, and glutathione‐disulphide reductase), and an increase in the production of ROS compared to the control in splenic neutrophils of tilapia. Collectively, our study provides evidence of the structure of tilapia NADPH oxidase genes and demonstrate that TCMBS application could modulate their activity in neutrophils to improve immunity in tilapia.     

MgO-induced defence against bacterial wilt disease in Arabidopsis thaliana

Bacterial wilt, caused by Ralstonia solanacearum, is a devastating soilborne disease in plants that limits the production of many crops worldwide. Although management of bacterial wilt has so far been unsuccessful, enhancing host resistance to the pathogen may be an effective control strategy. Recently, magnesium oxide (MgO) was found to induce defence responses against R. solanacearum in tomato plants. Here, the mechanisms underlying MgO-induced defence responses against R. solanacearum (MgO-i DARS) were investigated using Arabidopsis thaliana as a host plant. MgO-i DARS was confirmed in A. thaliana mutants deficient in jasmonic acid or ethylene signalling pathways as well as in the wildtype (Col-0) plants. In contrast, no MgO-i DARS was found in A. thaliana mutants deficient in the salicylic acid (SA) production (sid2-2) and signalling pathways (tga1-1 and npr-1). MgO treatment led to significant accumulation of SA in both roots and shoots of Col-0. The SA biosynthesis gene isochorismate synthase 1 (ICS1) was induced in roots and shoots of A. thaliana treated with MgO. An NADPH oxidase gene respiratory burst oxidase homolog D (AtRbohD) was up-regulated in both roots and shoots of Col-0 treated with MgO. No MgO-i DARS was observed in A. thaliana mutants deficient in AtRbohD. These results suggest that SA and RBOHD-mediated ROS are pivotal for MgO-i DARS in A. thaliana.     

Role of NOX1 in TNF-α-induced oxidative damage and inflammation in alveolar epithelial cells

AIM: To explore the role of NADPH oxidase 1 (NOX1) in tumor necrosis factor-α (TNF-α)-induced oxidative damage and inflammation in alveolar epithelial cells.METHODS: The mRNA and protein expression levels of NOX1 in alveolar epithelial cells after TNF-α treatment were determined by real-time PCR and Western blot. NOX1 siRNA and its negative control were transfected into the alveolar epithelial cells. After the induction of TNF-α, NOX1 levels in the cells were measured by real-time PCR and Western blot, and the content of malondialdehyde (MDA) in the cells was detected by thiobarbituric acid method. Xanthine oxidation assay was used to detect the activity of superoxide dismutase (SOD) in the cells. The contents of interleukin-4 (IL-4), IL-6 and IL-1β in cell culture medium were examined by ELISA. The rate of apoptosis was analyzed by flow cytometry. Western blot was used to detect the level of apoptotic protein cleaved caspase-3.RESULTS: The expression of NOX1 at mRNA and protein levels in TNF-α-induced cells was increased after induction (P<0.05). After transfection of NOX1 siRNA, the expression of NOX1 at mRNA and protein levels in the cell was downregulated (P<0.05). Transfection of siRNA negative control had no effect on the expression level of NOX1 in the cells. The content of MDA in the cells after TNF-α treatment was increased, the activity of SOD was reduced, the releases of IL-4, IL-6 and IL-1β by the cells were increased, and the apoptotic rate and the level of apoptotic protein cleaved caspase-3 were increased as compared with the cells that were not treated with TNF-α (P<0.05). The content of MDA in the cells with NOX1 knockdown induced by TNF-α was reduced, the activity of SOD elevated, and the releases IL-4, IL-6 and IL-1β, the apoptotic rate and the level of apoptotic protein cleaved caspase-3 decreased, as compared with the cells only treated with TNF-α induction (P<0.05).CONCLUSION: TNF-α induces the expression of NOX1 in the alveolar epithelial cells. Knockdown of NOX1 expression reduces cellular oxidative damage, releases of inflammatory factors, and cell apoptosis.     

FBW7 inhibits ox-LDL-induced granulosa cell apoptosis by promoting NOX1 degradation

AIM To investigate the effect of F-box and WD repeat domain containing protein 7 （FBW7） on the injury of granulosa cells （GCs） induced by oxidized low-density lipopretion （ox-LDL） stimulation and its potential mechanism. METHODS The GCs isolated from women of reproductive age with different obesity levels, the GCs isolated from high fatty diet （HFD）-fed rats, and the ox-LDL-treated rat GCs were collected, and the expression of FBW7 was detected by Western blot. After the rat GCs were infected with adenovirus encoding FBW7 （Ad-FBW7）, the cells were treated with ox-LDL （80 mg/L） for 48 h, and then the cell viability, apoptosis, NADPH oxidase （NOX） activity and the key subunit proteins of NOX were detected by CCK-8 assay, Annexin V/PI staining, lucigenin chemiluminescence and Western blot, respectively. The effect of FBW7 over-expression on NOX1 degradation was evaluated by cycloheximide （CHX） chase assay. RESULTS The expression of FBW7 was lower in the GCs isolated from obese women of reproductive age, the GCs isolated from HFD-fed rats, and the ox-LDL-treated rat GCs （P<0.05）. Over-expression of FBW7 inhibited ox-LDL-induced injury of GCs, NOX activity, and NOX1 expression （P<0.05）. The results of CHX chase assay showed that over-expression of FBW7 accelerated the degradation of NOX1 protein under ox-LDL condition （P<0.05）. CONCLUSION Over-expression of FBW7 reduces ox-LDL-induced injury of GCs by accelerating NOX1 degradation and then weakening NOX activity.     

亚精胺对渗透胁迫的玉米幼苗叶片NADPH氧化酶和抗氧化酶活性的影响

为了探讨多胺增强植物抗旱性的机理,研究了渗透胁迫下,玉米品种农大108(抗旱性较强)和掖单13号(抗旱性较弱)幼苗叶片中亚精胺(spermidine:Spd)含量与质膜NADPH氧化酶和3种抗氧化酶——超氧物歧化酶(SOD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性的变化.结果表明,渗透胁迫48 h,抗旱性弱的掖单13号幼苗叶片的NADPH氧化酶活性的上升幅度明显大于抗旱性强的农大108,而农大108幼苗叶片Spd含量以及SOD,CAT和APX活性的上升幅度明显大于掖单13号.外源Spd处理明显提高了掖单13号幼苗叶片的Spd含量以及SOD,CAT和APX活性,也明显抑制了NADPH氧化酶活性的上升.Spd的生物合成专一性抑制剂——甲基乙二醛-双(鸟嘌呤腙)(MGBG)处理,则明显抑制了渗透胁迫下农大108叶片中SOD,CAT和APX活性以及Spd含量的上升,也明显提高了NADPH氧化酶活性.结果还表明,渗透胁迫下玉米幼苗叶片的Spd可能通过抑制NADPH氧化酶活性以及促进抗氧化酶活性而降低叶片内的活性氧水平,从而减轻活性氧对幼苗的伤害.     

Effects of oxidative stress on high-fat,high-salt diet and sucrose solution-induced metabolic syndrome in nephropathy rats

AIM: To establish a suitable animal model of nephropathy associated with metabolic syndrome (MS) induced by abnormal diet, and to investigate the effects of oxidative stress on renal damage in MS rats. METHODS: Normal 7-week-old male SD rats were randomly divided into 2 groups.The animals were fed with normal chow (control group, n=10) or high-fat and high-salt diet plus 20% sucrose solution (MS model group, n=10) for 20 weeks. Systolic blood pressure (SBP) was measured monthly. The levels of blood glucose, serum and urinary creatinine (Cr), total cholesterol (TC), triglycerides (TG), fasting insulin (FIns), urinary protein, urinary albumin and urinary sodium were determined. Insulin resistance (HOMA-IR), creatinine clearance (Ccr), urinary protein excretion (UPE), urinary albumin excretion (UAE) and urinary sodium excretion (USE) were calculated. Renal total-antioxidant capacity (T-AOC), inhibiting superoxide anion capacity (ISAC), malondialdehyde (MDA) content, and antioxidant enzyme activity were measured. Renal protein expression of Cu/Zn-SOD, NADPH oxidase subunit p47^phox and p22^phox was detected by Western blotting. In addition, pathological changes of the kidney were observed with PAS and Masson staining,and degree of glomerulosclerosis (GS) and tubulointerstitial injury was evaluated. RESULTS: Compared with control rats, SBP, TC, TG, FIns, USE and UAE were increased in MS rats. Furthermore, the MS rats showed a significant elevation of renal MDA content, p47^phox protein expression and GS score, and reduction of T-AOC, ISAC, SOD activity, and Cu/Zn-SOD protein expression in the kidney. CONCLUSION: SD rats fed with abnormal diet produce a suitable animal model of MS nephropathy that mimics the major features of human MS. Oxidative stress caused by up-regulation of NADPH oxidase expression and down-regulation of SOD expression may be one of the mechanisms leading to MS renal damage.