Effect of AIR on Inflammatory Reaction Infected by Brucella

In order to investigate the effect of AIR on inflammatory reaction infected by Brucellamelitensis (16M), the AIR domain of Tecpr1 gene of murine macrophages RAW264.7 were knocked down (I-A), overexpressed (O-A) and reversed (OA-IA). Using the chlorine fluorescein (DCFH-DA) as a probe, we detected the variation of ROS production and mitochondria distribution by confocal laser scanning microscopy. We observed the expression changes of NLRP3, ASC and Caspase-1 by qRT-PCR and the expression changes of IL-18,IL-1β and Caspase-1 in host cells by ELISA. The results showed that 16M could stimulate RAW264.7 cells to produce ROS by time-dependent pathway, and I-A group and O-A group showed more abnormal accumulation of mitochondrial. The results of qRT-PCR and ELISA suggested that it had effect on the expression levels of NLRP3, ASC,Caspase-1 and IL-18, IL-1β and Caspase-1 in cells of different groups. Those results indicated that with AIR gene deletion, the release amount of ROS changed, mitochondrial clustered abnormally, and AIR was closely related to the activation of inflammasomes and induction of inflammatory reactions.     

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。

     

Antioxidant role of propolis extract against oxidative damage of testicular tissue induced by insecticide chlorpyrifos in rats

Pesticides induce oxidative stress leading to generate free radicals and alternate the antioxidant or oxygen free radical scavenging enzyme system. This study was conducted to investigate the oral toxicity of chlorpyrifos toward male rat and the oxidative stress of the sub-lethal dose (9 mg/kg; 1/25 LD₅₀) on the lipid peroxidation level (LPO), reduced glutathione content (GSH) and antioxidant enzymes; catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activities of testicular tissue. Also, the protective effects of propolis extract (50 mg/kg b.w.) alone or in combination with chlorpyrifos were investigated. The oral administration of chlorpyrifos significantly caused elevation in LPO level by 1.79-fold as compared to control. The activities of antioxidant enzymes including CAT, SOD, GPx and GST were decreased significantly (23.66%, 27.75%, 29.13% and 11.52%) as well as the level of GSH decreased by 21.97% in testicular tissue as compared to control animals. Co-administration of propolis extract with chlorpyrifos or alone in male rats decreased LPO level, normalized CAT, SOD GPx and GST activities, while GSH content was increased in testicular tissue. We conclude that propolis extract significantly reduces chlorpyrifos-induced oxidative stress in rat testis and the protective effect of the pre-treatment with propolis extract as attenuating agent could be due to its antioxidant properties.     

低温胁迫诱导斑马鱼ZF4细胞ROS及MAPK相关蛋白表达的影响

为了研究鱼类低温下分子调控机制及其与活性氧(reactive oxygen species,ROS)之间的关系,本实验对斑马鱼(Danio rerio)胚胎成纤维ZF4细胞进行不同程度的低温胁迫(18℃和10℃),监测其在不同低温胁迫时间下(1 d、3 d和5 d)ROS的变化以及丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路中蛋白的表达情况。结果显示:(1)DCFH-DA探针法表明在低温胁迫作用下,细胞内ROS含量增加。细胞内ROS上升水平与外界胁迫压力成正相关。低温处理3 d后,18℃和10℃的细胞,其ROS含量与对照组(28℃)相比分别显著升高到(1.23±0.04)倍(P0.05)和(2.31±0.08)倍(P0.05)。(2)Western Blot检测磷酸化的p38(p-p38)和磷酸化的JNK(p-JNK p54和p-JNK p46)的水平变化,结果显示低温胁迫可以使p38和JNK的活性增强,且均在10℃处理3 d达到最高水平。(3)进一步检测DNA断裂标记蛋白γH2A.X的表达水平,结果显示,无论在18℃还是10℃,其在第3天呈现高表达。本实验初步证实低温胁迫能够诱导斑马鱼细胞ROS的产生;通过对不同时间点的蛋白表达水平检测,发现p-JNK、p-p38和γH2A.X激活时间点与低温诱导ROS表达时间点相吻合。该研究为后期对斑马鱼细胞低温胁迫实验奠定基础,其中低温下处理3 d可以作为一个检测各个蛋白变化的关键时间点。     

植物谷胱甘肽应答非生物胁迫的分子机制

谷胱甘肽(GSH)是一种普遍存在于植物中的抗氧化剂,在维持组织抗氧化防御和调节氧化还原敏感信号转导中起着关键作用。深入研究GSH在非生物胁迫中的作用,对从分子水平揭示植物GSH积累的调控机制具有重要意义。本研究从植物GSH代谢途径及其相关酶、GSH在植物应激反应中的调节、GSH参与植物激素代谢等方面进行综述,并对谷胱甘肽在植物生长发育、与其它信号通路间交互作用的研究前景进行展望,以期为植物谷胱甘肽代谢以及其在非生物胁迫方面的研究提供一定的理论参考。     

DEHP对土壤蚯蚓氧化胁迫及DNA损伤的研究

土壤环境中的酞酸酯污染日益严重,为了探讨和分析典型酞酸酯邻苯二甲酸二(2-乙基己基)酯(Di(2-ethylhexyl)phthalate,DEHP)对土壤动物的生态毒理效应,以赤子爱胜蚓为指示生物,暴露于DEHP浓度为CK、0.1、1、10、50 mg kg~(-1)人工土壤中,并于染毒后的7、14、21、28d取样测定。通过蚯蚓体内的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物(POD)、谷胱甘肽转移酶(GST)等指标反映DEHP对蚯蚓的氧化激活程度,通过活性氧自由基(ROS)的含量反映DEHP对蚯蚓的细胞毒性,通过丙二醛(MDA)含量和Olive尾矩反映DEHP对蚯蚓的遗传毒性,从多个角度评价DEHP对蚯蚓的生态毒理效应。结果表明:(1)在DEHP的刺激下,各浓度组蚯蚓的SOD、CAT、POD、GST活性均呈激活状态,表明DEHP对蚯蚓的抗氧化酶有诱导作用;(2)DEHP影响蚯蚓的ROS含量,各染毒处理组与对照组相比均升高且差异性明显,表现出明显的剂量—效应关系;(3)对比各染毒处理组之间的数据,DEHP对蚯蚓MDA含量的影响无明显规律;(4)DEHP可使蚯蚓GST呈激活状态,表明中高浓度的DEHP对蚯蚓GST具有诱导作用;(5)DEHP能够引起蚯蚓体腔细胞DNA的损伤,且随着浓度的增加,Olive尾矩值随之增加,说明DNA损伤程度与DEHP浓度之间具有剂量-效应关系。从实验结果可以看出,DEHP可以对蚯蚓机体和DNA造成一定程度的损伤,表现出较强的生态毒理效应。     

Hepatoprotective activity of angiotensin-converting enzyme (ACE) inhibitors, captopril and enalapril, against paraquat toxicity

Paraquat is a highly toxic herbicide that is used in most countries without restriction. The cytotoxic action of paraquat is mediated by reactive radicals that are products of its metabolic reduction in cells. It has already been hypothesized that some angiotensin-converting enzyme inhibitors (e.g., captopril and enalapril) could show antioxidant and radical scavenging activity through their structural thiol groups, increasing antioxidant enzymes production or nitric oxide synthesis. In this study the hepatoprotective effect of captopril and enalapril against paraquat induced oxidative stress cytotoxicity was evaluated in isolated rat hepatocyte. Subtoxic concentrations of captopril (0.2 mM) and enalapril (0.2 mM) significantly (p < 0.05) protected the hepatocytes against paraquat (2 mM) induced oxidative stress cytotoxicity markers including: cell lysis, reactive oxygen species (ROS) generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Moreover, we showed that non-thiol enalapril acts as well as thiol containing captopril at inhibiting oxidative stress cytotoxicity markers. Finally, our results support the hypothesis that it is the increase in nitric oxide synthesis and not the presence of the thiol group that accounts for the antioxidant activity of ACE inhibitors.     

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.     

白藜芦醇对奶牛性控冻精品质和体外受精能力的影响

本研究旨在探究不同浓度白藜芦醇处理对奶牛性控冻精品质和体外受精能力的影响。在解冻后的奶牛性控冻精中分别添加0、10~(-3)、10~(-4)、10~(-5) mol/L的白藜芦醇,各组精子在受精液中获能孵育1.5 h后,测定精子质量和体外受精能力。结果表明:添加白藜芦醇均可有效降低性控冻精中活性氧(ROS)含量、提高顶体完整活精子比例(P0.05),其中10~(-3)、10~(-4) mol/L的白藜芦醇处理对降低ROS含量最为显著;10-4 mol/L的白藜芦醇处理可显著降低丙二醛(MDA)含量并提高性控冻精的卵裂率和囊胚率(P0.05)。综上所述,白藜芦醇作为一种外源性天然抗氧化剂,通过降低性控精液中过量的ROS水平、抑制精子脂质过氧化反应、保护顶体完整性,从而提高性控精子质量和体外受精能力。     

Accumulation of H<Subscript>2</Subscript>O<Subscript>2</Subscript> in xylem fluids of cucumber stems during ASM-induced systemic acquired resistance (SAR) involves increased LOX activity and transient accumulation of shikimic acid

Systemic acquired resistance (SAR) to Colletotrichum orbiculare was induced in young cucumber (Cucumis sativus) plants within 3 h of ASM (acibenzolar-S-methyl) application onto the first leaves. A potent signal associated with significant accumulation of hydrogen peroxide in xylem fluids from severed stems appeared to be rapidly translocated from elicited lower leaves within 3 h and 6 h after treatment. Some metabolites of the shikimate, phenylpropanoid and lignin biosynthetic pathways were quantified and significant increases in the levels of shikimic acid were observed in ASM-treated plants challenge-inoculated with the anthracnose fungus. Furthermore, the expression of the 5-enolpyruvylshikimate-3-phosphate synthase gene (EPSPS) was 1.5 times higher within 12 h after ASM treatment in challenge-inoculated plants than in the untreated control. The involvement of lipoxygenase activity, shikimic acid and others such as caffeic acid in the induction of SAR is discussed.