苦参碱对奶牛乳腺上皮细胞增殖、凋亡及抗氧化能力的影响

【目的】探究苦参碱对体外培养的奶牛乳腺上皮细胞(BMECs)增殖、凋亡及抗氧化能力的影响。【方法】利用含0(A组),25(B组),50(C组),75(D组)和100μg/mL(E组)苦参碱的培养基培养奶牛乳腺上皮细胞。通过四甲基偶氮唑盐(MTT)法检测BMECs活性,采用流式细胞仪(AnnexinV/PI双染法)检测苦参碱对BMECs凋亡的影响,并检测苦参碱对BMECs抗氧化酶活性及丙二醛(MDA)含量的影响,采用real-time PCR对BMECs中Caspase-3、p53、STAT1和SOCS3基因的相对表达量进行检测。【结果】用药5d时,低质量浓度(25和50μg/mL)苦参碱对BMECs增殖具有促进作用,高质量浓度(75和100μg/mL)苦参碱对细胞增殖具有抑制作用;B~E组BMECs的凋亡率均极显著高于A组(P0.01);B~E组BMECs培养上清液中NO和乳酸脱氢酶(LDH)水平明显高于A组。B~E组BMECs的过氧化氢酶(CAT)活性均比A组高,其中C组极显著高于A组(P0.01);B~E组的谷胱甘肽过氧化物酶(GSH-Px)活性均极显著高于A组(P0.01),E组的超氧化物歧化酶(SOD)水平极显著高于A组(P0.01),各组MDA含量无显著性差异。与A组相比,苦参碱上调了B~E组BMECs中Caspase-3、p53、STAT1和SOCS3基因的相对表达量。【结论】低质量浓度苦参碱能够促进BMECs增殖,高质量浓度苦参碱则会抑制BMECs增殖;不同质量浓度苦参碱均可提高BMECs的抗氧化能力,其中50μg/mL苦参碱提高BMECs抗氧化能力的效果最明显。     

果蔬籽油生物活性研究进展

果蔬籽油是由果蔬籽经压榨或萃取工艺制得的植物油脂。果蔬籽油含有丰富的维生素、多不饱和脂肪酸、有机酸等活性成分,这些成分赋予了果蔬籽油抗氧化、抗癌、调节血脂和调节免疫系统等生物活性。本文综述了果蔬籽油生物活性的研究进展,旨在为果蔬籽油的综合开发和利用提供理论依据和指导。     

Resistance monitoring and enzyme activity in three field populations of cowpea aphid (Aphis craccivora) from Egypt

The cowpea aphid (Aphis craccivora) is one of the most important sucking insect pests attacking certain legumes in Egypt particularly faba bean, cowpea and pea. In this study we monitored the resistance level of three field populations of A. craccivora to seven insecticides belonging to three different chemical classes (organophosphates, carbamates and neonicotinoids). The three populations were collected from three governorates in Egypt namely Dakahlia, Qalyobia and Beni Suef. Diagnostic concentrations (LC₉₀ values for susceptible strain) for each insecticide were established using a leaf dipping technique. Resistance monitoring showed that the field population from Dakahlia was highly resistant to all the tested insecticides. In a similar manner, the population from Qalyobia was also resistant to all insecticides except for fenitrothion to which only moderate resistance was observed. The field population from Beni Suef exhibited a lower level of resistance to all the seven tested insecticides.Biochemical assay showed that esterase activity in these three field populations was generally higher as the enzyme activity ratio ranged from 4.3 to 7.8 fold more than that for the susceptible strain. The activity of the other measured detoxifying enzymes (glutathione -S- transferase and mixed function oxidases) was moderate in the populations from Qalyobia and Dakahlia. Nevertheless, the enzyme activity in A. craccivora collected from Beni Suef was variable and differed slightly from the activity measured in the susceptible strain. Monitoring insecticide resistance among the three aphid populations was a proactive approach to detect any shift in insecticide efficiency. The possible occurrence of resistance in the cowpea aphid to the tested insecticides may be due to the higher activity of carboxylesterases. Further studies on the resistance mechanism to these insecticides are needed to provide insights in how to manage and delay the onset of the resistance and thus prolong the performance of insecticides against A. craccivora.     

固态发酵醋糟饲料对育肥猪生长性能、养分表观消化率、血清指标及粪便中挥发性脂肪酸含量的影响

本试验旨在研究固态发酵醋糟饲料(SFVD)对育肥猪生长性能、养分表观消化率、血清指标及粪便中挥发性脂肪酸(VFA)含量的影响。选择体况相近[(59.04±0.33)kg]的健康“杜×长×大”三元杂交生长育肥猪60头,随机分为2个组,每组3个重复,每个重复10头猪。对照组饲喂基础饲粮+150 mg/kg土霉素钙,试验组饲喂基础饲粮+5%的SFVD。预试期6 d,正试期35 d。结果表明:与对照组相比,基础饲粮中添加5%的SFVD对育肥猪平均日采食量(ADFI)具有一定的促进趋势(P=0.056);显著降低了育肥猪饲粮中干物质(DM)、粗蛋白质(CP)、有机物(OM)和总能(GE)的表观消化率(P<0.05),对粗脂肪(EE)表观消化率有一定的促进趋势(P=0.052);显著提高了育肥猪血清谷草转氨酶(AST)和谷胱甘肽过氧化物酶(GSH-Px)活性(P<0.05),血清高密度脂蛋白(HDL)和免疫球蛋白A(IgA)含量也呈现出一定的升高趋势(P=0.055和P=0.077);显著提高了育肥猪粪便中丙酸含量(P<0.05),粪便中甲酸和总挥发性脂肪酸(TVFA)含量也呈现出一定的升高趋势(P=0.071和P=0.054)。由此可见,基础饲粮中添加5%的SFVD对育肥猪生长性能、肠道健康及免疫性能的提高具有潜在的促进作用。     

单宁酸对生长猪胃-小肠仿生消化中消化酶活性及饲粮粗蛋白消化率的影响

本试验旨在探讨单宁酸对生长猪胃、小肠仿生消化中消化酶活性及玉米-豆粕型饲粮干物质和粗蛋白消化率的影响，为评价单宁酸的生物学效应提供参考。试验一采用单因素完全随机设计，考察在无饲粮下2种单宁酸对猪模拟胃液、模拟小肠液消化酶活性的影响。设5个处理，单宁酸添加量分别为0 mg （胃液体积为20 mL，小肠液体积为22 mL）；单宁酸1，10 mg；单宁酸1，20 mg；单宁酸2，10 mg；单宁酸2，20 mg。测定各处理的胃蛋白酶、淀粉酶、胰蛋白酶和糜蛋白酶的活性。试验二考察玉米-豆粕型饲粮添加单宁酸对猪仿生消化中胃、小肠阶段消化酶活性及养分消化率的影响。采用单因素完全随机设计，设5个处理，单宁酸在饲粮中的含量分别为0 mg·（2 g）^-1；单宁酸1，10 mg·（2 g）^-1；单宁酸1，20 mg·（2 g）^-1；单宁酸2，10 mg·（2 g）^-1；单宁酸2，20 mg·（2 g）^-1。测定仿生消化中胃阶段0.5和4 h时胃蛋白酶活性，小肠阶段0.5、4和8 h时淀粉酶、胰蛋白酶和糜蛋白酶活性及生长猪胃-小肠仿生消化测定饲粮的干物质和粗蛋白消化率。结果表明：1）无饲粮的情况下，和空白对照组相比，2种单宁酸对模拟胃液中胃蛋白酶活性无显著影响（P>0.05），单宁酸1比单宁酸2更高地降低了模拟小肠液中淀粉酶、胰蛋白酶和糜蛋白酶的活性（P<0.05）。2）在饲粮进行仿生消化的胃消化0.5～4 h内，除4 h时10 mg·（2 g）^-1添加量外，添加单宁酸1时胃蛋白酶的活性均显著高于添加单宁酸2时的相应值（P<0.05），除单宁酸2在消化0.5 h外，2种单宁酸在添加10 mg·（2 g）^-1时胃蛋白酶活性均显著高于20 mg·（2 g）^-1添加量的相应值（P<0.05）。在小肠仿生消化0.5 h时，饲粮中添加单宁酸1、2的2个水平对消化液中淀粉酶活性无显著影响（P>0.05），但均显著降低了糜蛋白酶的活性（P<0.05）；单宁酸1的消化液中胰蛋白酶活性高于单宁酸2的相应值（P<0.05）。在小肠仿生消化4 h时，除添加水平为20 mg·（2 g）^-1时的糜蛋白酶活性外，饲粮中添加单宁酸1消化液中淀粉酶、糜蛋白酶活性高于添加单宁酸2的相应值，而胰蛋白酶活性低于添加单宁酸2的相应值（P<0.05）。单宁酸1、2的两个添加量均降低了胰蛋白酶和糜蛋白酶的活性（P<0.05）。在小肠仿生消化8 h时，饲粮中单宁酸的添加量影响了淀粉酶的活性，但单宁酸1和单宁酸2各两个添加量在淀粉酶的平均活性上无显著差异（P>0.05）。单宁酸1、2的两个添加量均降低了胰蛋白酶、糜蛋白酶的活性（P<0.05）。3）与对照组相比，两种单宁酸在两种添加水平下均显著降低了饲料粗蛋白消化率（P<0.05），且单宁酸2比单宁酸1更多地降低了饲粮粗蛋白的消化率（P<0.05）。综上所述，在有、无饲粮条件下，单宁酸对消化酶活性呈现不一致影响。单宁酸影响饲粮粗蛋白的消化率可能主要与消化液中糜蛋白酶活性降低以及单宁酸与饲粮中的化学成分形成螯合物降低了小肠消化酶的水解效率有关。     

miR-106b-5p靶向KLF4调控山羊肌内前体脂肪细胞分化

旨在明确miR-106b-5p对山羊肌内前体脂肪细胞分化的影响，并确定这种作用是通过靶向KLF4来实现的。本研究利用实时荧光定量PCR （quantitative real-time PCR，qRT-PCR）技术检测miR-106b-5p在山羊肌内前体脂肪细胞分化过程中的表达模式，通过脂质体转染技术将miR-106b-5p mimic和miR-106b-5p inhibitor转入体外培养的山羊肌内前体脂肪细胞，油红O染色法从形态学验证miR-106b-5p对脂肪细胞中脂滴积聚的影响，qRT-PCR检测预测的靶标基因KLF4和脂肪分化标志基因的表达情况，利用双荧光素酶报告系统鉴定miR-106b-5p与KLF4的靶标关系。qRT-PCR结果显示，miR-106b-5p在山羊肌内前体脂肪细胞诱导分化第3天时表达量最高。在山羊肌内脂肪细胞中干扰miR-106b-5p后油红O染色显示脂滴聚积减少，过表达miR-106b-5p后脂滴聚积增加。在山羊肌内前体脂肪细胞中转染miR-106b-5p inhibitor后PPARγ表达量显著降低（P<0.05），而KLF4表达量极显著升高（P<0.01）；转染miR-106b-5p mimic后LPL和PPARγ表达量极显著升高（P<0.01）。荧光素酶活性试验结果显示，过表达miR-106b-5p可显著抑制KLF4荧光活性。miR-106b-5p通过靶向并负调节KLF4的表达促进山羊肌内脂肪细胞分化。     

不同处理对芹菜种子萌发过程中抗氧化系统及激素水平的影响

为研究芹菜种子的萌发机制,采用不同试剂组合（5 g/L NaOH+10% PEG、10% PEG+500 mg/L壳聚糖、5 g/L NaOH+500 mg/L壳聚糖）和不同温度(18、24、30℃)对芹菜种子进行处理,观察芹菜种子的萌发指标,并探究种子萌发过程中的抗氧化系统及内源激素水平的变化。结果表明,30℃对种子发芽率、发芽势及发芽指数提高具有促进作用,加快种子萌发进程。与蒸馏水对照组相比,试剂组合处理在18、24℃时种子发芽率、发芽势和发芽指数升高,促进种子萌发,18℃时5 g/L NaOH+500 mg/L壳聚糖处理下的发芽势最高,24℃时5 g/L NaOH+10% PEG处理下发芽势达到最大值;30℃时不同试剂组合处理间发芽率、发芽势和发芽指数无明显差异。18、30℃对种子具有一定程度的胁迫作用,种子SOD和CAT活性较24℃条件下有所增加;18℃时MDA含量和脯氨酸含量显著增加;5 g/L NaOH+10% PEG处理能显著降低MDA含量。萌发过程中,芹菜种子内部ABA含量下降,GA₃、ZA含量增加。试剂组合及适当高温有利于提高芹菜种子的发芽率、发芽势和发芽指数,促进种子萌发,提高种子萌发整齐度;芹菜种子萌发过程中抗氧化系统和激素水平会积极响应不同的处理条件。     

Effect of feeding different sources of selenium on growth performance and antioxidant status of broilers

ABSTRACT

1. This study was conducted to determine the effect of different sources of selenium (Se) on breast and liver tissue deposition, apparent metabolisable energy (AME), growth performance and antioxidant status of broilers, measured as Se content in liver and breast tissues and glutathione peroxidase (GSH-Px) in blood, when used in 0–35 d broiler chicken diets.

2. A total of 200 male Ross 308 broilers were used in the feeding trial, which comprised two dietary phases, a starter from 0 to 21 d and finisher from 21 to 35 d of age. Four treatments with 10 replications each were used. A control diet (C) was formulated that was sufficient in protein and energy (230 and 215 g/kg of crude protein and 12.67 and 13.11 MJ/kg of metabolisable energy, respectively), for both phases, but contained background Se only from the feed ingredients. Diet 2 (IS) was supplemented with 10.35 g/t inorganic, elemental source of Se. Diet 3 (SY) was supplemented with 136.36 g/t selenised yeast, an organic source derived from Saccharomyces cerevisiae. Diet 4 (SS) was supplemented with 0.666 g/t sodium selenite, an inorganic source.

3. Birds fed the SY diet consumed less and weighed less than those fed IS or C (P < 0.05; 0–35 d of age), but there was no difference compared to birds fed SS diets. There were no differences in FCR or dietary AME between broilers fed different Se sources. All diets containing supplementary Se increased concentrations in the liver and breast muscle, and for GSH-Px levels in blood compared to birds fed the C diet (P < 0.001). Birds fed SY diets had greater Se levels in liver and breast tissues compared to birds fed any of the other diets (P < 0.001).

4. Diets supplemented with Se had variable effects on broiler growth performances and antioxidant status. Feeding Se from a yeast source has higher transfer into breast tissues. Feeding different sources and levels of Se to birds in a more challenging situation to induce oxidative stress may bring more conclusive results.     

Fusarium solani Upregulated Sesquiterpene Synthase Expression,Sesquiterpene Production and Allelopathic Activity in Piper betle L.

Activators of sesquiterpene synthase (STS) gene expression and sesquiterpene production in Piper betle L. were examined using quantitative real time PCR and gas chromatography mass spectrometry methods, and the allelopathic activity of untreated and Fusarium solani-treated betel extracts was tested on seed germination and on the shoot and root growth of Thai rice variety PSL2 (Oryza sativa cv. Phitsanulok 2) and three dominant paddy weeds (Eclipta prostrata, Echinochloa crus-galli and Chloris barbata). The results demonstrated that F. solani dramatically upregulated STS expression and productions of β-cubebene, β-caryophyllene and germacrene D sesquiterpene when compared with the untreated control, and that betel extracts had a greater inhibitory effect on weeds than on rice. The effects were more clearly detected on seed germination and root growth than on shoot growth, and they were found to be dose-dependent. It is also noted that F. solani-treated extract had stronger effects than the untreated extract. The species most sensitive to the allelopathic effects was C. barbata, germination of which was completely inhibited even at a dose of 0.1 mg/mL untreated extract. With regards to rice, although betel extract at 1.0 mg/mL showed no inhibition on germination, it affected the elongation of rice roots, in addition to those of the tested weeds. The obtained data suggested that F. solani has potential as an activator of sesquiterpene allelochemical production via STS expression, the latter leading to the treated betel extract having a stronger phytotoxic effect. These results were beneficial in the promotion of natural herbicide production using biotechnology.     

Dietary clinoptilolite influences antioxidant capability and oxidative status of broilers

Natural clinoptilolite has been shown to have positive effects as an antioxidant, which means it traps free radicals in its complex structure, inactivating and eliminating them. Synthetic or modified clinoptilolite delays lipid peroxidation with water-soluble peroxyl radicals, and reduces the catalytic production of radicals to protect the organism. The objective of this study was to evaluate the effect of natural clinoptilolite and modified clinoptilolite on the antioxidant status of broilers. The antioxidant capability of natural clinoptilolite or modified clinoptilolite is exerted, at least in part, by increasing glutathione content in liver and intestinal mucosa, the superoxide dismutase, and glutathione peroxidase activity in serum, liver, and intestinal mucosa. In addition, to enhance broiler performance, there is a reduction of the concentration of malondialdehyde in serum, liver, and intestinal mucosa.