响应面法优化酶法制备藻蓝蛋白抗氧化肽的实验研究

为优化碱性蛋白酶酶解藻蓝蛋白制备抗氧化肽的工艺条件,采用响应面分析法,以超氧阴离子自由基清除率为响应值,研究了[E]/[S]、酶解温度和时间对制备抗氧化肽工艺的影响.结果表明,制备藻蓝蛋白肽的最佳酶解条件为:温度为44.9℃、时间为6.4 h和[E]/[S]为3.6%,此时对超氧阴离子自由基清除率达到75.39%.在该条件下制备的藻蓝蛋白酶解产物(0.5 g/L)还原能力的吸光度为1.03,并达到稳定.     

虾壳红色素的抗氧化和抗肿瘤作用

为综合利用虾、蟹等水产品壳废料,采用有机溶剂萃取方法提取虾壳中的红色素,它是一类以虾青素及其酯为主的类胡萝卜素。比较虾壳红色素与维生素Ｃ在抑制小鼠肝匀浆脂质过氧化,以及抑制人大肠癌ＳＷ１１１６细胞株增殖的作用。结果表明虾壳红色素对小鼠肝匀浆脂质过氧化具有明显的抑制作用,其效果比维生素Ｃ强１０倍以上;浓度为１×１０－９及１×１０－８ｍｏｌ／Ｌ虾壳红色素对人大肠癌ＳＷ１１１６细胞株增殖抑制率分别为２３．４％和３９．０％,在浓度为１×１０－７ｍｏｌ／Ｌ中培养的细胞基本死亡。提示了虾、蟹壳废料潜在的应用价值。     

镍添加对水鳖(Hydrocharis dubia)叶片镍和营养元素含量以及氧化胁迫和抗氧化系统的影响

采用营养液水培的方法,研究不同镍(Ni)添加浓度(0、0.05、0.10、0.15、0.20 mmol/L)对水鳖(Hydrocharis dubia)叶片中Ni元素积累、营养元素(Ca、Mg、K、Fe和Mn)含量、氧化胁迫和抗氧化系统的影响.结果显示:(1)叶片中Ni含量以浓度依赖的方式增加;(2)叶片中营养元素含量(Ca、Mg、K、Fe、Mn)也受影响,其中Ca、Mg含量随Ni浓度增加而显著上升;K含量随Ni浓度增加有所升高,但不显著;Mn含量虽在较大Ni浓度时有所下降,但处理组始终高于对照组;而Fe含量则随Ni浓度增加而下降;(3)Ni胁迫诱导了明显的氧化胁迫,主要表现为:O·-2产生速率以及过氧化氢和丙二醛含量显著升高,可溶性蛋白以及光合色素含量明显下降;(4)随着Ni浓度的升高,过氧化物酶活性逐渐升高,过氧化氢酶活性呈下降趋势,超氧化物歧化酶活性在Ni添加浓度为0.05 mmol/L时出现短暂升高随后下降;维生素E和黄酮含量随Ni浓度增加呈缓慢上升趋势;还原型谷胱甘肽未发生明显变化,非蛋白巯基和植物络合素含量均表现为先升后降的趋势;总抗氧化能力随Ni浓度升高先增强后减弱.由此可见,Ni添加引起水鳖叶片中Ni元素的累积和营养元素含量变化,同时产生明显的氧化胁迫,打破了抗氧化系统的平衡.     

pH胁迫对不同规格刺参生长及免疫酶活性的影响

以大、中、小3种不同规格的刺参(Apostichopus japonicus)为对象,在pH为7.3、7.5、8.2、8.7、8.9不同环境中养殖30d,观测刺参行为、生长及溶菌酶(LZM)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性。结果显示:当pH为7.5、8.7时, 3种不同规格刺参均能正常活动、摄食、生长,未表现出异常状况;若超出此范围,刺参出现粪便细短、残饵增多、摄食量减少等现象,尤其在pH 8.9时,中规格刺参有明显棘刺收缩的现象,小规格刺参不同程度地出现身体收缩、棘刺平滑状态,而大规格刺参无明显表观不适现象。不同pH对3种规格刺参的特定生长率(SGR)有显著影响,随着pH升高或者降低,刺参SGR均逐渐下降,与对照组差异显著(P0.05),规格越小刺参受到抑制作用越大;刺参LZM、SOD、CAT活性随pH胁迫时间的延长呈现先升高后下降的趋势,在第10天时达到最高,与对照组差异显著(P0.05),至30 d胁迫结束时,在pH 7.3、8.9条件下,刺参免疫酶活性显著低于对照组(P0.05),而pH为7.5、8.7时,刺参免疫酶活性与对照差异不显著; pH胁迫对不同规格刺参非特异性免疫酶活性的影响存在差异,在相同胁迫条件下,酶活性依次为大规格中规格小规格,当pH7.3、8.9时,小中大规格分别在胁迫20 d、25 d、25 d后免疫酶活性显著低于对照组(P0.05)。研究表明, pH胁迫会对刺参生长及免疫产生明显的影响,在刺参养殖过程中, pH属关键的理化因子之一,应密切关注其变化。     

Relationships of growth and mortality to enzymatic activity,and the relative mRNA expression of cultured scallops Patinopecten yessoensis in the Yellow Sea,China

The bottom aquaculture of yesso scallop (Patinopecten yessoensis) has developed rapidly in the north of Yellow Sea, China, but not without accompanying productivity problems. We, therefore, conducted field surveys to investigate factors related to growth and mortality rates of bottom-cultured scallops after release. Specifically, we focused on the effects of total antioxidant capacity (TAOC) and three key enzymes involved in antioxidant and metabolic function:glutamic pyruvic transaminase (GPT), lactic dehydrogenase (LDH), and superoxide dismutase (SOD). Across summer and winter, we also measured the relative mRNA expression of GPT and SOD to understand their seasonal variation in the scallops, along with how such variation correlated to growth and mortality. Results show that bottom cultured scallops experienced mass mortality during the first six months post-release. During winter (December), scallops grew more rapidly and suffered less mortality than in summer (August). The observed lower performance probably resulted from less advantageous environmental factors during the summer seasons, such as high temperature and low dissolved oxygen. These environmental stressors enhance protein consumption while decreasing energetic resources in scallops. Furthermore, scallops in summer exhibited high antioxidant levels that probably competed for energy with process integral to growth and survival. These negative factors combined to elevate mortality rates. In conclusion, we provided evidence suggesting correlations between metabolic/antioxidant activity and growth and mortality of bottom-cultured yesso scallops. These correlations implied us an accurate method to estimate the performance of bottom culture system. Suggestions about innovative aquaculture techniques were also discussed in the study. Our results might provide a possible guideline to the improvement of bottom culture techniques for this commercially valuable seafood species.     

海洋酸度变化下孔石莼对农药硫丹的生态响应研究

硫丹作为一种持久性有机氯农药,其对水生生物的毒性已引起广泛关注。本文以大型海藻孔石莼为材料,探究了海洋酸度变化下硫丹对孔石莼的生态毒理学效应。实验结果表明,低浓度硫丹对孔石莼的生长、光合色素及2种抗氧化酶均有明显的促进效应,50μg/L的硫丹促进效应最为显著,然而随着硫丹浓度的升高或暴露时间的延长,抑制作用明显增强。pH=7.9会促进孔石莼的生长,但会降低藻体内光合色素的含量。低pH环境会进一步加剧硫丹对孔石莼的毒性效应,两者的交互作用在不同时间段发挥不同的主效应。     

海草抗氧化系统及其对逆境胁迫的响应特征

海草具有独特的进化地位和重要的生态价值,广泛分布于潮间带和潮下带浅海海域,易遭受多种环境因子变化的威胁,抗氧化系统在海草抵御逆境胁迫的过程中具有非常重要的作用。本文综述了海草抗氧化系统的组成、特征及其对主要逆境胁迫的响应特征研究进展,阐述了海草主要酶促抗氧化机制,并将北半球代表性海草物种——鳗草抗氧化系统相关酶基因归类分析。目前对于海草逆境胁迫的研究主要集中于单一胁迫下主要抗氧化酶(如SOD、CAT、GST)及其转录组的变化,对于多胁迫因子协同作用和非酶抗氧化物及其他抗氧化酶的响应特征研究较少;另外对于逆境胁迫下不同海草种类间抗氧化系统和关键基因的响应差异也有待深入研究。     

小麦耐盐细胞系的筛选及其抗氧化能力分析

通过逐级提高氯化钠浓度的筛选方法获得了小麦耐盐细胞系。与对照细胞系相比 ,耐盐细胞系在盐分胁迫下能维持较高的超氧化物歧化酶、过氧化物酶和过氧化氢酶活性 ,同时具有较强的脂溶性抗氧化能力。说明在细胞水平上小麦对盐分耐受力的高低与其抗氧化能力的强弱密切相关。     

Adaptation of the antioxidant defence system in hydrothermal-vent mussels (Bathymodiolus azoricus) transplanted between two Mid-Atlantic Ridge sites

The vent mussel Bathymodiolus azoricus is the dominant member of the Northern Mid‐Atlantic Ridge (MAR) hydrothermal megafauna, and lives in an environment characterized by temporal and spatial variations in the levels of heavy metals, methane and hydrogen sulphide, substances which are known to increase reactive oxygen species levels in the tissues of exposed organisms. To evaluate the effects of two contrasting hydrothermal environments on the antioxidant defence system of this vent mussel species, a 2‐week transplant experiment was carried out involving mussels collected from the relatively deep (2300 m), and chemical rich, Rainbow vent field. These were transplanted to the shallower (1700 m), and relatively less toxic, Lucky Strike vent field. To achieve this objective, levels of superoxide dismutase, catalase (CAT), total glutathione peroxidase (GPx), selenium‐dependent glutathione peroxidase and lipid peroxidation (LPO) were measured in the gills and mantle tissues of resident and transplant mussels before and after the transplant experiment. With the exception of CAT, the gills of the transplanted mussels had significantly higher antioxidant enzyme activity compared with the basal levels in the donor (Rainbow) and recipient (Lucky Strike) populations; whereas the antioxidant enzyme levels in the mantle tissues of the transplants reflected the baseline levels of activity in the native Lucky Strike mussels after 2 weeks. In contrast, LPO levels were significantly higher in both tissue types in the transplants than in either the source or the recipient populations, which suggested a response to hydrostatic pressure change (note, the transplant animals were brought to the surface for transportation between the two vent fields). The fact that the Rainbow mussels survived the transplant experience indicates that B. azoricus has a very robust constitution, which enables it to cope behaviourally, physiologically and genetically with the extreme conditions found in its naturally contaminated deep‐sea environment.