高场强超声波对芒果多酚氧化酶和过氧化物酶钝化效果的研究

以芒果中多酚氧化酶(PPO)和过氧化物酶(POD)为研究对象,探讨加热与超声波作用对多酚氧化酶和过氧化物酶钝化效果的影响。结果表明:单独的加热(≤50℃)或0℃低温超声波(403.19、601.25、799.31 W/cm2)处理酶液,随着时间的延长,酶活性逐渐下降,但酶残留活性均在75%以上;而超声波与热处理结合能使POD和PPO显著失活,在45℃,799.31 W/cm2条件下处理15 min后,PPO、POD残余酶活力分别降至38.66%、14.43%。试验结果表明:超声波和热处理结合对芒果PPO和POD的钝化有协同效应。     

不同生育期卷丹百合的多酚积累特性及其抗氧化活性

为探究不同生育期卷丹百合各器官中多酚类物质的积累规律及其抗氧化能力，分别采用福林–肖卡法、NaNO2–AlCl3法、香草醛比色法、pH示差法测定不同生育期(现蕾期、花期、半枯期、全枯期)卷丹百合各器官(基生根、鳞茎、茎生根、茎秆、叶片、株芽、花）中总酚、总黄酮、总黄烷醇和花色苷的含量，运用DPPH法、铜离子还原法和金属螯合能力、抑制脂质过氧化活性，分析其多酚提取物的抗氧化能力。结果表明：不同生育期卷丹百合各器官多酚类物质的含量及其抗氧化活性存在差异；卷丹百合植株在花期总酚含量最高，半枯期的次之，全枯期的最少；花期，花中总酚含量最高，达14.75 mg/g，叶片中的总酚含量次之，珠芽的最低，仅为3.12 mg/g；卷丹百合多酚提取液的抗氧化能力在花期最强，其次是半枯期；相关性分析表明，卷丹多酚类物质与抗氧化活性在各时期、各器官呈现出显著(P<0.05)或极显著(P<0.01)的相关性，相关系数为–1.00~1.00，其中，花期和半枯期卷丹多酚类物质与铜离子还原力基本呈显著或极显著正相关。     

Immune defense enzymes: Advances in L‐amino acid oxidase of marine animals

L‐amino acid oxidase (LAAO) is able to stereoselectively catalyse the deamination of L‐amino acids to α‐keto acids, ammonia and H₂O₂. It has been reported that this enzyme has biological functions including inhibition of bacteria and antiviral, anti‐parasitic and anti‐tumour activities. To date, marine animal LAAO are constantly being identified. L‐amino acid oxidase derived from marine animals has been purified for Sebastes schlegelii, Platichthys stellatus, Gadus morhua, Signanus oramin, Aplysia californica and some others. In this paper, we review the reported progress of research on marine animal LAAO, including basic characteristics, tissue distribution and biological function. We intended to provide a reference for the further study of LAAO in marine animals.     

兰州百合多酚提取工艺优化研究

以多酚得率为优化指标,对兰州百合中多酚的提取工艺进行研究,通过单因素试验考查了溶剂类型、料液比、溶剂体积分数、超声功率、提取温度和提取液pH值对提取多酚得率的影响,在此基础上,为得到最优的兰州百合多酚提取条件,对乙醇浓度、提取温度、提取液pH值和超声功率4因素进行正交试验,结果得出兰州百合多酚提取最佳工艺参数组合为:乙醇浓度80%、提取温度40 ℃、提取pH值7、超声功率500 W,其多酚的萃取量为100.27 mg/g。     

5种蔷薇科树种多酚氧化酶比较基因组学分析

蔷薇科植物中有多种具有重要价值的水果,如苹果、梨、桃、草莓和黑树莓等。这些水果普遍容易发生由多酚氧化酶(PPO)介导的酶促褐变而导致重大经济损失,从全基因组角度分析比较PPO基因家族,有助于加深对PPO基因家族和功能的认识。采用比较基因组学的方法,对这5种植物的PPO基因家族进行了基因鉴定、染色体定位、编码蛋白的亚细胞定位、内含子统计分析、基因系统进化、基因倍增与丢失等特征分析。从这5种植物中,共计鉴定出了42个PPO基因,其中6个基因被认定是假基因;绝大多数基因编码的蛋白定位于叶绿体,2个定位于线粒体,3个是分泌型蛋白;PPO基因在染色体上有串联重复和散布两种形式;9个基因具有内含子,聚类结果显示可以将它们分为6个类型,每个基因类型在进化过程中都发生过基因倍增和丢失;同型基因内含子的位置和大小具有相似性。这些结果揭示蔷薇科植物PPO基因内含子是伴随着基因倍增产生的,基因倍增也是推动PPO基因多样化的重要动力,PPO基因倍增和丢失差异导致PPO基因数量在不同物种之间产生差异。     

红茶提取物对高尿酸血症小鼠血尿酸的影响

研究红茶提取物对高尿酸血症小鼠血尿酸的影响。将36只雄性KM小鼠随机分为空白组、模型组、红茶提取物低、中、高剂量组及别嘌呤醇组。空白组和给茶组连续1周分别灌胃生理盐水和红茶提取物,给茶组第7天造模后1 h给茶;模型组在第7天腹腔注射氧嗪酸钾并灌胃酵母膏造模。测定结果显示:与模型组相比,各给茶组血尿酸(UA)水平均降低;与模型组相比,给茶组血尿素氮(BUN)水平均降低,其中、高剂量给茶组BUN水平显著降低(P0.05),高剂量组差异极显著(P0.01);各给茶组血肌酐(Cr)值与模型组相比极显著下降(P0.001)。高剂量组黄嘌呤氧化酶(XOD)活性较模型组显著降低(P0.05),低、中剂量组有一定的抑制作用,但无显著差异。研究表明,红茶提取物对氧嗪酸钾和酵母膏导致的小鼠高尿酸血症有明显的改善作用。     

橄榄多酚氧化酶基因(PPO)克隆及其试管苗离体保存过程中的表达分析

以来源于橄榄成熟胚的橄榄试管苗为材料,进行PPO基因克隆,比较试管苗不同保存阶段PPO基因转录水平和PPO酶活性的变化。结果表明:克隆获得1个PPO基因全长cDNA序列,命名为PPO2,GenBank登录号为JQ319005,cDNA全长序列为1 934 bp,推测出橄榄PPO开放阅读框(ORF)1 818 bp,编码606个氨基酸。橄榄PPO2属于碱性、亲水、非分泌蛋白,可能定位于线粒体基质空间,具有3个典型的保守结构域,丝氨酸磷酸化占主导地位,并且二级结构以不规则卷曲为主。而三级结构预测表明,橄榄试管苗PPO2蛋白主要由α螺旋和不规则卷曲组成;进化树分析显示橄榄PPO蛋白与胡杨PPO关系最近,而与葡萄和莲的亲缘关系较远;qPCR分析显示,PPO2基因在橄榄试管苗不同保存阶段都有表达,但在橄榄试管苗保存1个月幼叶比较多时PPO2基因的表达量最高;而橄榄试管苗在分别保存1、2、3、4、5个月时PPO活性变化趋势不明显,当保存时间到6、7、8个月时PPO活性急剧增加。因此,PPO基因可能与幼嫩组织的启动分化有关,并在橄榄试管苗生长发育过程中发挥重要作用。     

Sulodexide protects against hyperglycemia-induced retinal vascular injury via suppressing NOX4/NLRP3 pathway

AIM To investigate the effect of sulodexide (SDX) on high glucose-induced damage in retinal microvascular endothelial cells. METHODS (1) High-fat diet combined with intraperitoneal injection of streptozocin were used to induce type 2 diabetes mellitus (DM) followed by injection of saline or SDX in C57BL/6J male mice. Retinal microvascular leakage and density, and the protein levels of NLRP3 inflammasome-related proteins, zonula occludens-1 (ZO-1) and NADPH oxidase 4 (NOX4) were measured. (2) Human retinal microvascular endothelial cells (HRMECs) were treated with normal glucose or high glucose with or without SDX, and were further transfected with siRNA to knock down NOX4, or infected by adenovirus to over-express NOX4. The protein levels of ZO-1, VE-cadherin (VE-Cad), NOX4 and NLRP3 inflammasome-related proteins as well as the level of reactive oxygen species (ROS) were detected. RESULTS Treatment with SDX increased the protein level of ZO-1, attenuated retinal leakage and NLRP3 inflammasome activation, and enhanced the density of microvasculature and the number of ganglion cells in diabetic retinas. The protein levels of ZO-1 and VE-Cad were decreased, while the levels of NOX4, NLRP3 inflammasome-related proteins and ROS generation were increased in high glucose-treated HRMECs. Silencing of NOX4 inhibited high glucose-induced increases in NLRP3 inflammasome and ROS generation, and decreases in the protein levels of ZO-1 and VE-Cad. Over-expression of NOX4 significantly increased the levels of NLRP3 inflammasome-related proteins and ROS generation in HRMECs, and reduced the protein levels of ZO-1 and VE-Cad. Treatment with SDX partly reversed NOX4 over-expression-induced changes. CONCLUSION SDX alleviates hyperglycemia-induced retinal microvascular endothelial injury via inhibiting NOX4/ROS/NLRP3 pathways.     

Effect of sulphide in Pacific white shrimp Penaeus vannamei under varying oxygen and pH levels

Sulphide is one of the known environmental stressors, which potentially binds to cytochrome C oxidase (COX), a key enzyme in the electron transport chain, thereby blocking oxygen transport and ATP production. To ascertain the toxic effects of sulphide on Pacific white shrimp, Penaeus vannamei, two distinct exposure experiments were carried out with varying concentration of sulphide (0–1 mg/L), dissolved oxygen (normoxia and hypoxia) and pH values (8.2, 6 and 5). The activity of enzymes viz COX, superoxide dismutase (SOD), phenoloxidase and lactate accumulation was investigated. Outer membrane integrity and COX monomer separation were also done with the isolated crude mitochondrial preparations. Results indicated a significant reduction (p ≤ .05) in COX enzyme activity in sulphide exposed treatments when compared to control. The reduction was more intense when pH levels were reduced under hypoxia condition. Lactate accumulation as a result of anaerobic metabolism was found to be higher in hypoxic treatments. No significant difference (p ≥ .05) was observed in superoxide dismutase activity between the treatments, whereas phenoloxidase activity significantly decreased at higher concentration sulphide. Separation of mitochondrial proteins resulted in the identification of ~205 kDa of COX monomer, and significant damage was found in outer membrane integrity under hypoxia and pH treatments. From this study, it is evident that at a given concentration, sulphide is toxic to P. vannamei, and in association with hypoxia and low pH, they further intensify sulphide toxicity. Our results indicated that sulphide toxicity should not be considered as a single factor, rather it should be a considered as combination of factors.