ROS对氯化镉诱导的人乳腺癌MCF-7细胞凋亡的影响

为研究活性氧(ROS)对氯化镉(CdCl2)诱导的人乳腺癌MCF-7细胞凋亡的影响,本文采用荧光探针DHR检测CdCl2诱导的MCF-7细胞内ROS的变化水平;分别采用MTT法和DNA Ladder法检测CdCl2对细胞的毒性作用和细胞DNA的凋亡情况;通过加入自由基清除剂LNAC抑制细胞内的ROS水平,从而研究ROS对细胞凋亡的影响。结果显示,1.00×10-3 mol/L的CdCl2处理MCF-7细胞6h,细胞内ROS水平为对照组的2.04倍,细胞存活率为26.54%,DNA出现片段化分解。用CdCl2+LNAC复合处理MCF-7细胞6h,细胞内ROS水平为对照组的1.25倍,细胞存活率为61.17%,DNA不出现凋亡条带。因此,由CdCl2所导致的MCF-7细胞凋亡和DNA片段化分解,与细胞内ROS水平的升高有关。     

In vitro effect of recombinant human tumor necrosis factor-alpha on canine neutrophil apoptosis

Neutrophils (polymorphonuclear leukocytes: PMNs) are essential for the host defense against various infections and are often injurious to the host, causing inflammatory diseases where tumor necrosis factor-alpha (TNF-alpha) is suggested to play an important role. Since an effect of TNF-alpha on canine PMN apoptosis has not been studied, canine PMNs were stimulated with recombinant human (rh)TNF-alpha in the present study to investigate the effect of TNF-alpha on canine PMN apoptosis. PMN apoptosis and function to produce ROS were assessed by flow cytometry. Delayed apoptosis was observed in the PMNs treated with rhTNF-alpha at 100 ng/ml, accompanied by retention of capability to produce ROS. However, PMN apoptosis was accelerated by rhTNF-alpha combined with cycloheximide. Therefore, it is indicated that TNF-alpha is able to activate anti- and pro-apoptotic pathways in PMNs and that the inhibition of PMN apoptosis by TNF-alpha requires protein synthesis in the PMNs.     

漆酶活化处理条件对竹材活性氧类自由基变化的影响

采用电子自旋共振波谱检测技术,分析漆酶活化毛竹材产生的自由基种类,并结合单因素方差分析方法,分析漆酶活化处理条件对毛竹材产生的活性氧类自由基相对浓度的影响。未处理毛竹材产生的自由基属酚氧自由基,而漆酶处理毛竹材产生的自由基属典型的活性氧类自由基。酶用量、反应体系pH值、处理时间、处理温度及Cu~(2 )浓度,均对活性氧类自由基相对浓度有极显著影响。当采用酶用量为30u/g、反应体系pH值为4、处理时间为2h、处理温度为60℃、Cu~(2 )浓度为30mmol/L时,漆酶活化毛竹材产生的活性氧类自由基的相对浓度最高。     

不同生境中半红树植物抗氧化防御研究

选取5种半红树植物,研究了其在不同生境(潮间带和陆生)抗氧化防御系统的变化。结果表明,陆生的黄槿、许树O2.产生速率和莲叶桐H2O2含量均显著的高于其潮间带居群,从而使这3种半红树叶片中抗氧化酶类(SOD、APX、CAT、POD和GR)活性均高于其潮间带居群。而处于潮间带杨叶肖槿、阔苞菊(除SOD外)的SOD、APX、CAT活性显著高于其陆生居群,此时,H2O2含量要显著低于其陆生居群,而陆生的阔苞菊居群SOD、POD、GR活性高于其潮间带居群,因此,O2.产生的速率显著低于其潮间带居群。不同树种对不同生境有着不同抗氧化防御策略,总的趋势是,陆生的半红树抗氧化酶活性均要高于其潮间带居群,尤其是POD、GR活性在5种半红树中均显著高于其潮间带居群,暗示环境胁迫造成的选择压力导致半红树植物适应性的反应。高温、高光照、干旱是对陆生半红树的逆境胁迫,因此,上调抗氧化酶活性,以降低逆境对半红树的膜脂过氧化,减少电解质外渗,最终提高了半红树的抗逆性。     

外源信号物质对肉苁蓉种子萌发与吸器形成POD、CAT活性及ROS含量的影响

利用外源信号物质氟草敏(norflurazon)和2,6-二甲氧基对苯醌(2,6-DMBQ)分别诱导肉苁蓉种子萌发与吸器形成,研究它们在此过程中对过氧化物酶(POD)、过氧化氢酶(CAT)活性及活性氧(ROS)含量的影响。结果表明：在诱导肉苁蓉种子萌发时,经norflurazon处理0~168 h后,种子中POD和CAT活性呈现显著升高的变化趋势,种子中ROS含量显著升高。在诱导肉苁蓉种子萌发体吸器形成时,经2,6-DMBQ处理0~72 h后,肉苁蓉种子萌发体中POD和CAT活性呈现显著升高的变化趋势,萌发体中ROS含量显著升高。表明在诱导肉苁蓉种子萌发与吸器形成过程中,外源信号物质norflurazon和2,6-DMBQ能影响POD、CAT活性及ROS含量的变化。

     

外源海藻糖调节西瓜细胞渗透胁迫抗性的研究

为探究外源海藻糖对渗透胁迫下西瓜细胞生长的影响,本研究以西瓜悬浮培养细胞为试材,测定外源海藻糖预处理后,渗透胁迫对西瓜悬浮培养细胞生长量、细胞外pH、细胞内ROS（活性氧簇）相对含量和微管骨架的影响。结果表明：渗透胁迫下西瓜细胞的生长量明显受到抑制,并且渗透胁迫可诱导西瓜悬浮培养细胞质外体碱化,ROS迸发,细胞微管骨架发生解聚;外源添加海藻糖可在一定程度上缓解渗透胁迫对西瓜悬浮细胞生长量的抑制作用,调节pH、ROS表达水平、并维持微管骨架结构的完整性。以上研究结果表明渗透胁迫下,海藻糖对西瓜细胞具有维持细胞生长、保护亚细胞结构并调节抗性反应的功能。     

Changes in antioxidative system and membrane damage of lettuce in response to salinity and boron toxicity

Individual and combined effects of salinity and B toxicity on growth, the major antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX) activities, ascorbic acid, proline, and H₂O₂ accumulation, and stomatal resistance (SR), malondialdehyde (MDA), membrane permeability (MP) and the concentrations of sodium (Na), chloride (Cl) and boron (B) of lettuce were investigated. Boron toxicity and salinity reduced growth of lettuce plants. Under B toxicity, B concentration of the plants was increased, but in the presence of NaCl, the concentration of B was significantly reduced. Sodium and Cl concentrations were increased in B + NaCl and NaCl treatments. Membrane damage was more pronounced in NaCl and B + NaCl treatments. Stomatal resistance of the plants was significantly increased by salinity treatments. The accumulation of proline and ascorbic acid was the highest in the B + NaCl treatment. In general, stress conditions significantly increased H₂O₂ and antioxidant enzyme (SOD, CAT and APX) activities. The present results indicate that stomatal closure is an important response of lettuce against NaCl and B + NaCl stress. Furthermore NaCl and B + NaCl toxicity-induced oxidative stress in lettuce resulting in lipid peroxidation and membrane damage. Increased antioxidant enzyme activities and also accumulation of ascorbic acid and proline are involved in order to overcome B- and NaCl-induced oxidative stress.     

植物-植食性昆虫互作关系中早期信号事件研究进展

诱导防御反应的产生取决于植物对植食性昆虫相关信号迅速和准确地识别。在植物与植食性昆虫互作过程中,早期信号事件是负责植物识别和触发下游信号转导途径的最早反应,它们发生在植物防御相关基因表达和防御相关代谢物产生之前,对植物的防御过程至关重要。本文将植物与植食性昆虫互作的早期事件,包括植物对植食性昆虫的感知、电信号和Ca~(2+)信号的产生和转导,活性氧的迸发以及促细胞分裂原活化蛋白激酶(MAPK)信号级联途径等研究进展进行了综述,并提出今后的研究重点与方向,促进对植物诱导防御反应调控网络的研究,以期为改进农业害虫的治理提供重要的理论和技术支撑。     

First Report of a Peroxiredoxin Homologue in Jellyfish: Molecular Cloning,Expression and Functional Characterization of CcPrx4 from Cyanea capillata

We first identified and characterized a novel peroxiredoxin (Prx), designated as CcPrx4, from the cDNA library of the tentacle of the jellyfish Cyanea capillata. The full-length cDNA sequence of CcPrx4 consisted of 884 nucleotides with an open reading frame encoding a mature protein of 247 amino acids. It showed a significant homology to peroxiredoxin 4 (Prx4) with the highly conserved F-motif (⁹³FTFVCPTEI¹⁰¹), hydrophobic region (²¹⁷VCPAGW²²²), ¹⁴⁰GGLG¹⁴³ and ²³⁹YF²⁴⁰, indicating that it should be a new member of the Prx4 family. The deduced CcPrx4 protein had a calculated molecular mass of 27.2 kDa and an estimated isoelectric point of 6.3. Quantitative real-time PCR analysis showed that CcPrx4 mRNA could be detected in all the jellyfish tissues analyzed. CcPrx4 protein was cloned into the expression vector, pET-24a, and expressed in Escherichia coli Rosetta (DE3) pLysS. Recombinant CcPrx4 protein was purified by HisTrap High Performance chelating column chromatography and analyzed for its biological function. The results showed that the purified recombinant CcPrx4 protein manifested the ability to reduce hydrogen peroxide and protect supercoiled DNA from oxidative damage, suggesting that CcPrx4 protein may play an important role in protecting jellyfish from oxidative damage.     

线粒体中活性氧的产生途径和测定方法

活性氧(ROS)是自然界普遍存在的含氧化合物的总称,对生物体的生理和细胞稳态的调节具有重要意义.但活性氧的测定方法仍然是生物研究领域的一个难点,本文综述了线粒体中活性氧的产生途径和测定方法.