活性氧(ROS)对小鼠早期胚胎胚细胞分裂的影响

在体外CZB培养的不同时期添加过氧化氢(2.1μmol/L),发育至囊胚阶段,制作胚胎标本,观察其囊胚发育率、胚细胞数和分裂相数。结果显示:0~12 h组和0~72 h组的胚胎发育率显著低于其他几组(P<0.05),对照组、12~24 h组、24~36 h组、36~48 h组之间均差异不显著(P>0.05);各组之间的胚细胞数和分裂指数均无显著差异。说明胚胎在体外培养时,2细胞期胚胎对外源性H2O2最敏感,也最容易发生发育阻滞。但是经H2O2处理之后,度过2细胞期发育阻滞的胚胎能在以后的发育中发生补偿性生长,其胚细胞数和分裂指数都和体外正常发育的胚胎没有差异。     

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.     

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.     

萝卜清除DPPH自由基活性研究

[目的]研究萝卜清除DPPH自由基的活性.[方法]通过对萝卜叶水提物、水提物不同极性部位萃取物、萝卜中脂肪油类有效成分β-谷甾醇进行清除DPPH（1,1-二苯基-2-三硝基苯肼）自由基活性测试,对萝卜的抗氧化能力进行评价.[结果]以柠檬酸为对照其清除能力大小顺序为：乙酸乙酯层＞二氯甲烷层＞萝卜叶水提物＞β-谷甾醇＞石油醚层＞柠檬酸.[结论]萝卜具有较好的抗氧化能力.     

模拟胃液条件下假蒟叶提取物抑制亚硝化反应的研究

[目的]研究不同溶剂的假蒟叶提取物对亚硝化反应的影响。[方法]在模拟胃液条件下,测定假蒟叶提取物亚硝酸盐清除率及亚硝胺合成阻断率。[结果]不同溶剂的假蒟提取物均有一定的抑制亚硝化反应作用,其中乙酸乙酯提取物的抑制作用最强,且其对亚硝酸盐的清除率及对亚硝胺合成的阻断率在一定的浓度范围内随其浓度的增加而增加,对亚硝酸盐清除能力较强,其IC50值为0.425mg/ml,对亚硝胺合成阻断的能力相对较低,其IC50值为4.064 mg/ml。[结论]假蒟叶提取物具有较好的亚硝化反应抑制活性且主要通过清除底物亚硝酸盐来实现。     

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。     

Antioxidant Activity of Pigment Extracted from Green-Wheat-Bran

The anthocyanin pigment extracted from green-wheat-bran was studied to identify its antioxidant activity.The antioxidant activities of the pigment were evaluated by anti-lipid peroxidation,total antioxidant activity （TAA）,superoxide anion radical scavenging activity （SARSA）,active oxygen scavenging activity （AOSA）,and DPPH （1,1-diphenyl-2 picrylhydrazyl free radical） radical scavenging activity.The results showed that the pigment had higher antioxidant activity and TAA,SARSA,AOSA and DPPH.scavenging activities at a certain concentration than Vc （antiscorbutic vitamin,vitamin C）,and the capacity increased with the increase of pigment concentration.Its TAA was 51.06 U mL-1,1.73 times of Vc,and SARSA 18 025.21 U mL-1,2.26% higher than Vc,and AOSA 3 776.31 U mL-1,1.24 times of Vc.As to the DPPH.scavenging activity of the pigment,there was a trend that higher concentration performed higher activity significantly improved with the company of Vc.The pigment showed significant antioxidant activities evaluated by different assays.Results will provide a better understanding on antioxidant activity of green wheat and allow the screening or breeding of green wheat varieties with higher antioxidant activity for food processing.     

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。     

茶蒸青叶中儿茶素类的制备及其对氧自由基的清除作用

以茶鲜叶经过蒸汽杀青得到的“蒸青叶”为原料，制备儿茶素类产物。采用HPLC分析了儿茶素类的含量和组分，同时采用邻二氮菲-Fe^2＋氧化法与NBT光还原法，研究了儿茶素类产物对两种氧自由基的清除能力。结果表明：儿茶素类产物具有较强的清除自由基的能力。在本实验条件下，其清除两种氧自由基的IC50分别为42．29mg／L（羟基自由基），5．99mg／L（超氧自由基），均低于从成品绿茶中制备的茶多酚。HPLC分析结果表明，儿茶素类产物的纯度为92．52％，其中酯型儿茶素占84．78％，这是其表现出强烈的清除自由基能力主要原因。从“蒸青叶”中分离制备儿茶素类的产率、纯度与活性均优于从成品绿茶中制备的茶多酚。     

除草剂苯噻草胺胁迫对水稻活性氧清除系统的影响

采用室内培养实验方法,对水稻幼苗的多项生理生化指标进行了测定。结果表明,随着除草剂苯噻草胺浓度的升高,水稻株高变矮,根长缩短;蛋白质、叶绿素和GSH含量应激升高。活性氧·O-2、H2O2和膜脂过氧化产物MDA的含量增加,组织自氧化速率增大,表明苯噻草胺对水稻幼苗的毒害作用主要是破坏了体内的活性氧平衡。水稻叶片中的CAT酶,根中的SOD和POD酶对苯噻草胺的浓度比较敏感。SOD与MDA含量呈显著正相关,提示了抗氧化酶清除活性氧的作用滞后于活性氧对膜脂过氧化的诱导作用。