PrP106—126诱导小胶质细胞BV-2抗氧化性能的变化

小胶质细胞活化是朊病的病理学特征之一。朊蛋白多肽PrP106—126具神经毒性,是研究异常腕蛋白（PrPSc）的理想工具。为探讨PrP106—126对小胶质细胞氧化压力的影响。本研究以小胶质细胞BV-2为细胞模型,PrP106—126作用48h,应用MTT和流式细胞仪检测细胞的活化情况,应用分子探针技术对细胞的氧化压力（reactiveoxygenspecies,ROs）进行检测,并通过荧光定量RT—PCR对与R0s相关的酶的mRNA表达进行了测定。结果表明PrPl06—126显著促进小胶质细胞BV-2的活化,并提高胞内的ROS水平;定量RT—PCR显示,PrP106—126显著降低细胞S0D-1（P〈0．01）表达水平、提高胞内Cat（P〈0．01）的表达水平;对Grx、Trx-1、和Trx-2mRNA的表达水平有升高的趋势,但未达到显著水平（P〉0．05）,对SOd-2、GPx、GR无显著性影响（P〉0．05）。从分子水平初步阐明小胶质细胞ROS升高的机理。     

两个玉米自交系对纹枯病的抗病反应机制初探

玉米纹枯病是由立枯丝核菌(Rhizoctonia solani Kühn)引起的一种重要病害。具有优良性状的自交系是开展玉米抗病品种选育的重要种质资源。本试验以西南地区两个优良玉米自交系R08与18-599R为材料,采用人工接种R.solani菌丝融合群AG1-IA,对成株期玉米进行了抗性鉴定,并对病原菌诱导后苗期玉米叶片上的活性氧(ROS)、抗病相关基因(PR1、ZmDREB2A、ZmERF3和WIP1)表达及细胞坏死情况进行了动态检测。结果显示,R08对纹枯病表现为中抗,而18-599R为高感材料;受R.solani侵染后,玉米叶片ROS的积累在R08中较18-599R多;两个自交系中基因表达量及表达时间存在显著差异;R08叶片出现显著的细胞坏死反应,而18-599R相对较轻。本研究表明两个自交系对R.solani的抗性差异主要体现在相关抗病反应的时间和强度上,这为进一步研究玉米纹枯病抗病机理提供了依据。     

Potassium fertilization improves apple fruit (Malus domestica Borkh. Cv. Fuji) development by regulating trehalose metabolism

Potassium (K) fertilization and antioxidant enzymes both positively influence plant growth and development. However, it is not known whether K treatment improves fruit development via increasing soluble sugar. In this study, K-treated apple (Malus domestica Borkh.) fruit was harvested from 60 to 150 days after full bloom and was analyzed for ROS production and trehalose metabolism. The results show that K fertilization improved fruit firmness, increased growth according to several parameters, increased soluble sugar content, and decreased ROS production. The ascorbate metabolic pathway more effectively reduced ROS production than catalase and peroxidase (POD) did under K treatment. Trehalose-treated fruit also showed higher activity of ascorbate-related enzymes (DHAR, GR, and APX) compared with non-treated fruit. The changes of antioxidant enzyme activity in trehalose-treated fruit corresponded to those in K-treated fruit. Moreover, trehalase (TREH) activity in fruit was notably reduced by K treatment. This demonstrates that K influences ROS production via regulating trehalose content and TREH activity in fruit. This study provides new insight into the K mechanism which improves fruit development, including fruit firmness and size.     

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.     

Effect of Psammaplin A on in vitro Development of Bovine Aging Oocytes

To investigate the effect of histone deacetylation inhibitor Psammaplin A (PsA) on the development of bovine aging oocytes in vitro,oocytes were randomly divided into control group,aging group and 50 mmol/L PsA treated aging group (PsA group).Immunofluorescence staining and JC-1 were used to detect the blastocyst rate of bovine oocytes after parthenogenetic activation,the number of cells in blastocysts,apoptosis,reactive oxygen species (ROS),glutathione (GSH) and mitochondrial membrane potential intensity of embryos.The results showed that the blastocyst rate of the aging group was significantly lower than that of PsA and control groups (P<0.05).The blastocyst rate of PsA group was not significantly different from that of control group (P>0.05).The number of cells in the blastocysts of control group and PsA group were significantly higher than that of aging group (P<0.05).The number of cells in the blastocysts of PsA group was not significantly different from that of control group (P>0.05).The apoptosis rate in aging group was significantly higher than that of control and PsA groups (P<0.05),the apoptosis rate of PsA group was significantly higher than that of control group (P<0.05).The GSH level of MⅡ oocytes in aging group was significantly lower than that of control and PsA groups (P<0.05).There was no significant difference in GSH level between control and PsA groups (P>0.05).The ROS level of the embryos in aging group was significantly higher than that of control and PsA groups (P<0.05).The ROS level in PsA group was significantly higher than that of control group (P<0.05).The mitochondrial membrane potential of early embryos of aging group 4-8 cells was significantly lower than that of control and PsA groups (P<0.05).The mitochondrial membrane potential intensity of control group was significantly higher than that of PsA group (P<0.05).In summary,PsA could effectively delay the aging of bovine oocytes and improve the quality of oocytes.     

稀释液中添加海藻糖对猪冷冻精液质量参数和活性氧的影响

本试验旨在探讨海藻糖对猪精子冷冻效果以及精子内活性氧(ROS)的含量影响。试验分对照组和4个海藻糖处理组(0.025,0.05,0.1和0.2 mol/L)。精子冷却后,添加0.1 mol/L海藻糖处理组精子活力显著(P<0.05)高于对照组,而精子内和稀释液中各组间ROS发生量都没有显著的差异。精子冷冻解冻后,添加0.05 mol/L海藻糖处理组的精子活力显著(P<0.05)高于对照组;添加0.05 mol/L和0.1 mol/L海藻糖处理组的精子生存率显著(P<0.05)高于对照组;添加0.1 mol/L海藻糖处理组顶体完整的精子百分比显著(P<0.05)高于对照组;4个海藻糖处理组膨胀精子百分比都显著(P<0.05)高于对照组;添加0.025 mol/L海藻糖处理组ROS水平显著(P<0.05)低于对照组。结果表明:海藻糖对精子冷冻保存是有益的,且能抑制精子内ROS的发生,但它的作用机制有待于进一步的研究。     

Characterization of semen collected by pharmacologically induced ejaculation from a stallion with seminal vesiculitis

The present study compared the quality of sperm collected by artificial vagina or pharmacologically induced ejaculation from a 10-year-old thoroughbred stallion with seminal vesiculitis. The pharmacological protocol involved intravenous administration of detomidine (0.01 mg/kg) and oxytocin (20 IU) and successfully induced ejaculation in all attempts of semen collection. Sperm motility, plasma membrane and acrosome integrity (PMAI), reactive oxygen species (ROS) levels, polymorphonuclear neutrophil (PMN) percentage, and bacterial profiles of fresh and cooled semen (5°C for 24 hr) were evaluated. Semen obtained by the pharmacological method presented reduced seminal volume, decreased PMN percentage and superior sperm motility in cooled samples. Moreover, higher PMAI and lower ROS levels were observed in semen collected by the pharmacological method. Therefore, pharmacologically induced ejaculation is an alternative to obtain semen with minimal contamination and with sperm of superior quality and longevity from stallions with seminal vesiculitis.     

In vitro effect of recombinant human tumor necrosis factor-alpha on canine neutrophil apoptosis

Neutrophils (polymorphonuclear leukocytes: PMNs) are essential for the host defense against various infections and are often injurious to the host, causing inflammatory diseases where tumor necrosis factor-alpha (TNF-alpha) is suggested to play an important role. Since an effect of TNF-alpha on canine PMN apoptosis has not been studied, canine PMNs were stimulated with recombinant human (rh)TNF-alpha in the present study to investigate the effect of TNF-alpha on canine PMN apoptosis. PMN apoptosis and function to produce ROS were assessed by flow cytometry. Delayed apoptosis was observed in the PMNs treated with rhTNF-alpha at 100 ng/ml, accompanied by retention of capability to produce ROS. However, PMN apoptosis was accelerated by rhTNF-alpha combined with cycloheximide. Therefore, it is indicated that TNF-alpha is able to activate anti- and pro-apoptotic pathways in PMNs and that the inhibition of PMN apoptosis by TNF-alpha requires protein synthesis in the PMNs.     

植物-植食性昆虫互作关系中早期信号事件研究进展

诱导防御反应的产生取决于植物对植食性昆虫相关信号迅速和准确地识别。在植物与植食性昆虫互作过程中,早期信号事件是负责植物识别和触发下游信号转导途径的最早反应,它们发生在植物防御相关基因表达和防御相关代谢物产生之前,对植物的防御过程至关重要。本文将植物与植食性昆虫互作的早期事件,包括植物对植食性昆虫的感知、电信号和Ca~(2+)信号的产生和转导,活性氧的迸发以及促细胞分裂原活化蛋白激酶(MAPK)信号级联途径等研究进展进行了综述,并提出今后的研究重点与方向,促进对植物诱导防御反应调控网络的研究,以期为改进农业害虫的治理提供重要的理论和技术支撑。