NR和NOS在CTK延缓离体小麦叶片衰老中的作用

用一氧化氮(NO)清除剂血红蛋白(Hb)、硝酸还原酶(NR)抑制剂钨酸钠(Na2WO4)、一氧化氮合酶(NOS)抑制剂L-硝基精氨酸甲酯(L-NAME)并结合激动素(KT)和玉米素(ZT)两种细胞分裂素(CTK)处理离体小麦叶片,测定分析各处理的相关生理生化指标,以明确NR和NOS在CTK延缓离体小麦叶片衰老过程中的作用.结果显示:KT和ZT单独处理均能显著延缓离体小麦叶片衰老过程中叶绿素、可溶性蛋白含量的降低,抑制丙二醛(MDA)的积累,促进NR和NOS活性升高;在Hb、Na2WO4或L-NAME存在时,上述KT和ZT延缓衰老的效应均显著减弱,同时NR和NOS活性的升高也分别被Na2WO4和L-NAME显著抑制.该结果暗示CTK延缓离体小麦叶片衰老可能与其诱导了NR和NOS活性的提高,进而促进NO的生成有关.     

一氧化氮参与外源谷胱甘肽对盐胁迫下番茄幼苗抗氧化损伤的调控

采用营养液水培,研究外源谷胱甘肽(GSH)对NaCl和NaCl+Hb(牛血红蛋白,一种一氧化氮清除剂)处理下番茄(Lycopersicon esculentum)幼苗叶片中一氧化氮合酶(NOS)活性及其一氧化氮(NO)水平、膜脂过氧化程度、活性氧(ROS)积累和ROS清除能力的影响。结果表明:施用Hb使盐胁迫下番茄幼苗叶片的氧化损伤程度加剧,NO含量下调,但对GSH含量无显著影响。外源GSH的施用显著提高了盐胁迫下番茄幼苗叶中内源GSH与NO水平,单脱氢抗坏血酸还原酶(MDHAR)和硝酸还原酶(NR)活性,超氧化物歧化酶(SOD)活性(除72 h),过氧化氢酶(CAT)活性(除48 h),处理48和72 h的谷胱甘肽还原酶(GR)、抗坏血酸过氧化物酶(APX)和脱氢抗坏血酸还原酶(DHAR)活性;降低电解质渗透率、硫代巴比妥酸(TBARS)含量以及处理48和72 h的过氧化氢(H_2O_2)和超氧阴离子(O_2ˉ·)含量。外源喷施GSH亦显著提高NaCl+Hb处理下番茄叶中抗坏血酸(AsA)、GSH和NO水平,CAT和APX活性,处理24和48 h的NR活性以及处理48和72 h的NOS活性;降低电解质渗透率及H_2O_2和TBARS含量。表明外源GSH通过介导内源GSH和NO水平上调,提高抗氧化酶活性和ROS的清除能力来缓解NaCl和NaCl+Hb处理导致的氧化损伤。因此,NO参与了外源GSH对盐胁迫下番茄幼苗抗氧化损伤的调控。     

外源NO供体对小麦离体叶片过氧化氢代谢的影响

分析了外源一氧化氮 (nitricoxide ,NO)供体硝普钠 (sodiumnitroprusside ,SNP)对离体小麦 (TriticumaestivumL .)叶片过氧化氢 (H2 O2 )含量及其清除酶活力的调节作用。不同浓度的SNP (1mmol/L和 5mmol/L)处理 3 0min内 ,离体小麦叶片H2 O2 含量均有一个显著上升的过程 ,同时过氧化物酶 (POD)活力受到显著抑制 ,而过氧化氢酶 (CAT)活力则轻微下降 ;处理 3 0min到 2 4 0min时 ,POD活力的抑制状态基本维持不变 ,而CAT活力开始恢复上升 ,H2 O2 含量也相应地开始下降。粗酶液的体外实验也表明 ,SNP对POD和CAT的抑制类型不同 ,前者可能是不可逆抑制 ,后者则可能是可逆抑制。因此NO可通过对POD和CAT的不同抑制作用来调节小麦叶片内源H2 O2 含量     

燕麦叶片衰老与活性氧代谢的关系

燕麦连体叶片与高体叶片衰老中,过氧化氢酶和超氧物歧化酶(SOD)活性下降,脂类过氧化产物丙二醛(MDA)迅速积累,组织自动氧化速率显著加快。植物激素BA,GA_3,2,4—D及光、亚胺环己酮(CH),EDTA处理均不同程度地延缓离体叶片的衰老过程,同时抑制过氧化氢酶和SOD活性下降,阻止MDA的积累和组织自动氧化速率的提高.推测叶片衰老中活性氧起着重要的作用。     

施氮量对小麦叶片硝酸还原酶活性、一氧化氮含量和气体交换的影响

研究了不同施氮量对冬小麦分蘖到抽穗期叶片硝酸还原酶(NR)活性、一氧化氮(NO)含量、气体交换参数和籽粒产量的影响.结果表明:叶片光合速率（P_n）、蒸腾速率（T_r）、瞬时水分利用效率（IWUE）和产量均随施氮量的增加呈先升高后降低的趋势,在180 kg·hm^-2氮处理时达到最高.随施氮量的增加,叶片NR活性提高; 在分蘖期和拔节期,叶片NR活性与NO含量呈显著线性相关（R²≥0.68,n=15）,NO含量和气孔导度（G_s）呈显著正二次相关（R²≥0.43,n=15）;低氮处理下,NR活性较低使叶片NO含量维持在较低水平,促进气孔开放,高氮处理下,NR活性较高使叶片NO含量增加,诱导气孔关闭;在抽穗期叶片NR活性和NO含量无显著相关关系,虽然NO含量和G_s也呈显著正二次相关（R²≥0.36,n=15）,但不能通过施氮提高NR活性来影响叶片NO含量,进而调节叶片气孔行为.合理施氮使小麦叶片NO含量维持在较低水平,可提高叶片G_s、T_r和IWUE,增强作物抗旱能力,促进光合作用,提高小麦产量.     

外源NO供体对小麦离体叶片过氧化氢代谢的影响

分析了外源一氧化氮（nitric oxide ,NO）供体硝普钠（sodium nitroprusside,SNP）对离体小麦（Triticum aestivumL.）叶片过氧化氢（H2O2）含量及其清除酶活力的调节作用。不同浓度的SNP (1 mmol/L和5 mmol/L) 处理30 min内, 离体小麦叶片H2O2含量均有一个显著上升的过程, 同时过氧化物酶（POD） 活力受到显著抑制,而过氧化氢酶（CAT）活力则轻微下降;处理30 min到240 min时,POD活力的抑制状态基本维持不变,而CAT 活力开始恢复上升, H2O2含量也相应地开始下降。粗酶液的体外实验也表明,SNP对POD和CAT的抑制类型不同,前者可能是不可逆抑制,后者则可能是可逆抑制。因此NO可通过对POD和CAT的不同抑制作用来调节小麦叶片内源H2O2含量。     

6-苄氨基嘌呤对离体小麦叶片衰老及蛋白质降解过程的影响

6-苄氨基嘌吟(6—BA)对离体小麦叶片游离氨基酸累积的阻抑作用与呼吸抑制剂对叶片游离氨基酸累积的影响不同。从可溶性糖(蔗糖和葡萄糖)放射自显影图谱的比较看到在6—BA处理与对照之间没有阴显差异。6—BA显著地抑制了离体叶片蛋白质分解酶活力的增加。在促进离体叶片蛋白质降解过程的条件下,6—BA的作用才表现得明显。用聚丙烯酰胺凝胶电泳分析离体叶片的可溶性蛋白质,看到6—BA能够降低叶片中部分-Ⅰ蛋白质在离体后的丢失。 综上所述,认为6-苄氨基嘌呤延缓离体叶片衰老可能主要是通过阻抑离休叶片蛋白质的降解。叶绿体可能是6—BA延缓离休叶片衰老的主要作用部位。     

小麦叶片衰老进程中呼吸作用和ATP含量变化(简报)

小麦离体叶片在水中暗下衰老时出现呼吸高峰,体内ATP量也逐渐上升,到达高峰后再下降,两者的变化相平行。KT处理延迟呼吸高峰来临,ABA处理呼吸高峰提前。在光下衰老或用KT处理时,叶片内ATP水平较暗下或水中低。光下以及KT处理中,即延迟衰老作用最明显时,体内ATP水平最低。     

铀在小麦幼苗中的积累分布及其对叶片光系统活性的影响

分别采用不同浓度的铀[0、5、20、50、100mg.L-1 UO2(NO3)2.6H2O]对五叶期的‘西科麦3号’小麦幼苗于水培条件下处理7d,分析小麦对铀的吸收积累情况,并通过快速叶绿素荧光诱导动力学OJIP曲线及820nm光吸收曲线,分析铀对叶片光系统Ⅱ(PSⅡ)和光系统Ⅰ(PSⅠ)活性的影响。结果表明:(1)小麦对铀的富集系数和转移系数较小,吸收的铀主要集中在根部。(2)铀胁迫显著降低了小麦叶片捕光色素叶绿素b的含量,并显著影响小麦叶片两个光系统的活性;铀显著抑制PSⅡ反应中心的活性,但是对PSⅡ的电子供体侧和受体侧电子传递活性及PSⅠ的活性则表现为促进作用。(3)低浓度的铀处理会影响小麦叶片中两个光合系统之间的平衡,对PSⅠ性能的促进作用显著大于PSⅡ。     

精胺与亚精胺对光照下玉米离体叶片光合羧化酶活性的影响（简报）

玉米离体叶片在光下衰老时,其RuBPC、PEPCase和PPDK（丙酮酸磷酸二激酶）活性逐渐降低。精胺（Spm）和亚精胺（Spd）可阻止这三种酶的活性下降,还可阻止叶绿素和可溶性蛋白含量下降,延缓光下玉米离体叶片的衰老。     

Combined NO and PG inhibition augments alpha-adrenergic vasoconstriction in contracting human skeletal muscle

Sympathetic alpha-adrenergic vasoconstrictor responses are blunted in the vascular beds of contracting muscle (functional sympatholysis). We tested the hypothesis that combined inhibition of nitric oxide (NO) and prostaglandins (PGs) restores sympathetic vasoconstriction in contracting human muscle. We measured forearm blood flow via Doppler ultrasound and calculated the reduction in forearm vascular conductance in response to alpha-adrenergic receptor stimulation during rhythmic handgrip exercise (6.4 kg) and during a control nonexercise vasodilator condition (using intra-arterial adenosine) before and after combined local inhibition of NO synthase (NOS; via N(G)-nitro-L-arginine methyl ester) and cyclooxygenase (via ketorolac) in healthy men. Before combined inhibition of NO and PGs, the forearm vasoconstrictor responses to intra-arterial tyramine (which evoked endogenous noradrenaline release), phenylephrine (a selective alpha1-agonist), and clonidine (an alpha2-agonist) were significantly blunted during exercise compared with adenosine treatment. After combined inhibition of NO and PGs, the vasoconstrictor responses to all alpha-adrenergic receptor stimuli were augmented by approximately 10% in contracting muscle (P <0.05), whereas the responses to phenylephrine and clonidine were also augmented by approximately 10% during passive vasodilation in resting muscle (P <0.05). In six additional subjects, PG inhibition alone did not alter the vasoconstrictor responses in resting or contracting muscles. Thus in light of our previous findings, it appears that inhibition of either NO or PGs alone does not affect functional sympatholysis in healthy humans. However, the results from the present study indicate that combined inhibition of NO and PGs augments alpha-adrenergic vasoconstriction in contracting muscle but does not completely restore the vasoconstrictor responses compared with those observed during passive vasodilation in resting muscle.     

Fiat Lux: selective delivery of high flux of nitric oxide (NO) to biological targets using photoactive metal nitrosyls

In addition to its beneficial roles in blood pressure regulation, immune response, neurotransmission, and redox balance, nitric oxide (NO) can induce cellular apoptosis at relatively high concentrations. Since photoactive metal nitrosyls can deliver NO under the control of light, they are uniquely suited as NO drugs in photodynamic therapy (PDT) to destroy cancer cells. Stable and photoactive metal nitrosyls can first be placed in close proximity of a malignant site and then triggered via pulses of light to deliver high flux of NO. During the past few years, a number of such metal-based 'NO carriers' have been synthesized and tuned for rapid release of NO upon exposure to UV or visible light. Using various chromophore conjugation strategies, attempts are now being made to photosensitize the M-NO bond to infrared light. Progress has also been made in incorporating metal nitrosyls into biocompatible matrices for site-specific delivery of NO to tumors. A combination of light and NO could offer a viable treatment modality for cancer.     

A molecular hybrid producing simultaneously singlet oxygen and nitric oxide by single photon excitation with green light

Combination of photosensitizers (PS) for photodynamic therapy with NO photodonors (NOPD) is opening intriguing horizons towards new and still underexplored multimodal anticancer and antibacterial treatments not based on “conventional” drugs and entirely controlled by light stimuli. In this contribution, we report an intriguing molecular hybrid based on a BODIPY light-harvesting antenna that acts simultaneously as PS and NOPD upon single photon excitation with the highly biocompatible green light. The presented hybrid offers a combination of superior advantages with respect to the other rare cases reported to date, meeting most of the key criteria for both PSs and NOPDs in the same molecular entity such as: (i) capability to generate ¹O₂ and NO with single photon excitation of biocompatible visible light, (ii) excellent ¹O₂ quantum yield and NO quantum efficiency, (iii) photogeneration of NO independent from the presence of oxygen, (iv) large light harvesting properties in the green region. Furthermore, this compound together with its stable photoproduct, is well tolerated by both normal and cancer cells in the dark and exhibits bimodal photomortality of cancer cells under green light excitation due to the combined action of the cytotoxic ¹O₂ and NO.     

Effect of Reducing Nitric Oxide in <Emphasis Type="Italic">Rumex</Emphasis> K-1 Leaves on the Photoprotection of Photosystem II Under High Temperature with Strong Light

The purpose of this study was to explore the effect of reducing nitric oxide (NO) in Rumex K-1 leaves on the photoprotection of photosystem II (PSII) under high temperature with strong light. Reducing the content of NO in Rumex K-1 leaves significantly aggravated the PSII photoinhibition and net degradation of D1 protein under high temperature with strong light, but not under high temperature in the darkness. The reduction of NO remarkably inhibited the electron transport of PSII in the leaves under high temperature and strong light, which resulted in an increase in excitation pressure and an over-accumulation of reactive oxygen species (ROS). The over-accumulation of ROS further damaged PSII. However, when the synthesis of D1 protein was inhibited, the D1 protein content and PSII activity were no longer influenced by reducing NO content in the leaves. The reduction of NO in leaves decreased the activities of ROS scavenger enzymes after treatment with high temperature and strong light for 2 h, which enhanced the over-accumulation of ROS to damage photosynthetic apparatus severely. All of these results suggest that NO was involved in the synthesis of D1 protein. Maintaining physiologically appropriate NO content in leaves will alleviate net degradation of D1 protein under high temperature with strong light to keep photosynthetic electrons flowing smoothly, which mitigates the accumulation of ROS in photosystems to avoid damage to the photosynthetic apparatus. Therefore, NO plays an important role in maintaining higher PSII photosynthetic performance under high temperature with strong light.     

Involvement of nitric oxide in light-mediated greening of barley seedlings

When seedlings are grown in the dark, proplastids of the developing leaf differentiate into etioplasts. Greening of etiolated plastids is stimulated by light, which is sensed by various types of photoreceptors. Nitric oxide (NO) has been shown to be a bioactive molecule that could take part in this light-mediated process in plants. In this paper, we show that emission of NO in barley seedlings increased concomitantly with increasing activities of nitric oxide synthase (NOS) during the greening. Treatment with sodium nitroprusside (SNP), a NO donor, increased the accumulation of chlorophyll contents, enhanced the accumulation of thylakoid membrane proteins, such as light harvesting complex of photosystem II (LHCII) and PSIA/B, and then improved the effective quantum yield of photosystem II (PSII) (Phi(PSII)) in the light. Instead, treatment with either NO scavenger 2-phenyl-4,4,5,5-tetramentylimidazoline-1-oxyl-3-xide (PTIO) or NOS inhibitor N(omega)-nitro-l-arginine (l-NNA) retarded the greening of etiolated-seedlings. Moreover, sodium ferrocyanide, an analog of SNP, nitrite and nitrate, two NO-decomposition products did not have any effect on the greening process. These results indicated that NO, as an endogenous signaling molecule, participates in light-mediated greening of barley seedlings, and exogenous NO accelerates this process.     

Online electrochemical monitoring of nitric oxide during photodynamic therapy

Photodynamic therapy (PDT), as a novel treatment modality, is based on the use of a photosensitizing agent with an excitation light source for the treatment of various malignancies. Its effect is mediated through reactive oxygen species and nitric oxide (NO), which are shown to be present in apoptosis. Individual differences among patients and even in different areas of the same tumor in one patient may cause a major problem with PDT: dose calculation during application of the light. An electrochemical sensor is proposed for online monitoring of NO generation as a solution of this problem. 5-Aminolevulinic acid (ALA) was administered as the photosensitizer in rat cerebellum. An amperometric sensor, selective to NO, was designed and tested both in vitro and in vivo during PDT. ALA-mediated PDT resulted in rapid generation of NO, starting as early as the application of light on the tissue. Simultaneous amperometric recordings have been carried out for 5 min during PDT. The progressive increase in NO concentration peaked at 1.10 min and then the response current began to decrease until it reached a plateau at around 70% of its peak value. This study, for the first time, electrochemically demonstrates the generation of NO during PDT. Rapid and stable responses obtained by the experimental setup confirmed that this method could be used as an online monitoring system for PDT-mediated apoptosis.     

Physiological,Pathological and Pharmacological Interactions of Hydrogen Sulphide and Nitric Oxide in the Myocardium of Rats with Left Ventricular Hypertrophy

Left ventricular hypertrophy (LVH) is characterized by increased myocardium thickness due to increased oxidative stress and downregulation of cystathione γ lyase (CSE) endothelial nitric oxide synthase (eNOS). Upregulation of CSE by hydrogen sulphide (H₂S) and ENOS by L-arginine can arrest the progression of LVH individually. The present study explored the combined treatment of H₂S and NO in the progression of LVH, and demonstrated that the response is due to H₂S, NO or formation of either new molecule in physiological, pathological, and pharmacological in vivo settings of LVH. Exogenous administration H₂S+NO in LVH significantly reduced (all p < 0.05) systolic blood pressure (SBP) and mean arterial pressure (MAP), LV index, heart index and oxidative stress when compared to the LVH group. There was downregulation of CSE mRNA and eNOS in the heart, and exogenous administration of H₂S+NO groups upregulated eNOS MRNA while CSE MRNA remained downregulated in the hearts of the LVH group. Similar trends were observed with concentrations of H₂S and NO in the plasma and tissue. It can be concluded that combined treatment of LVH with H₂S and NO significantly ameliorate the progression of LVH by attenuating systemic hemodynamic and physical indices, and by decreasing oxidative stress. Molecular expression data in the myocardium of LVH depicts that combined treatment upregulated eNOS/NO while it downregulated CSE/H₂S pathways in in vivo settings, and it is always eNOS/NO pathways which play a major role.     

Enhancement of apoptosis by nitric oxide released from alpha-phenyl-tert-butyl nitrone under hyperthermic conditions

The aim of this study was to examine whether a neuroprotector, PBN (alpha-phenyl-tert-butyl nitrone), enhances apoptosis induced by hyperthermia, which generates superoxide (O2-) intracellularly, since the release of nitric oxide (NO) from PBN under oxidative stress has been reported. When human myelomonocytic lymphoma U937 cells were treated with hyperthermia (44 degrees C, 10 min) and PBN, an increase in the concentration of nitrite in the culture medium, and a decrease in the hyperthermia-induced production of O2- was observed. Imaging using a fluorescence dye for intracellular NO, diaminofluorescein-2 diacetate (DAF-2 DA), revealed the formation of NO in the apoptotic cells treated with hyperthermia and PBN combined. Apoptotic endpoints were significantly enhanced by the combined treatment: a decrease in mitochondrial trans-membrane potential, cleavage of Bid, release of cytochrome c, and activation of caspase-8 and -3. An increase in the intracellular Ca2+ concentration ([Ca2+]i), externalization of Fas, and decrease in Hsp70 and phosphorylated HSF1 were observed following the combined treatment. Furthermore, scavengers of NO an d ONOO- significantly inhibited the enhancement of apoptosis, the externalization of Fas and the increase in [Ca2+]i. These results suggest that, (1) NO is released from PBN by hyperthermia, and subsequently reacts with O2- to form ONOO-, (2) NO and ONOO- are involved in the enhancement of apoptosis through Fas-mitochondria-caspase and [Ca2+]i-dependent pathways, and (3) a decrease in Hsp70 and phosphorylated HSF1 also contributed to the enhancement of apoptosis.