Role of vascular NAD(P)H oxidase in vascular remodeling

NAD(P)H oxidase was initially found in phagocytes and it participates in the generation of reactive oxygen species(ROS). Recent researches have showed that NAD(P)H oxidase also expresses in other tissues including blood vessels and it plays a critical role in vascular remodeling through ROS which are important signaling molecules in vascular cells.This article reviews the biochemical characterization, activation paradigms, structure, and function of this enzyme.     

Effect of hypercapnia on lysyl oxidase-dependent collagen cross-linking and hypoxia-induced pulmonary hypertension in rat

AIM: To investigate the effect of hypercapnia on hypoxia-induced pulmonary hypertension and the changes of lysyl oxidase (LOX) and extracellular matrix collagen cross-links in the rat. METHODS: Sprague-Dawley rats were randomly divided into 4 groups:normoxia group, hypoxia group, hypercapnia group and hypoxia+hypercapnia group. LOX activity was detected by fluorescence spectrophotometry. LOX protein expression was detected by immunohistochemistry and Western blot. The mRNA expression of LOX in the pulmonary artery was detected by real-time PCR. RESULTS: The levels of mean pulmonary artery pressure (mPAP), RV/(LV+S) and WA/TA in hypoxia group were significantly higher than those in normoxia group (P<0.01). Moreover, the levels of mPAP and RV/(LV+S) in hypoxia+hypercapnia group were significantly lower than those in hypoxia group (P<0.01). However, no significant difference of mPAP and RV/(LV+S) between hypercapnia group and normoxia group was observed. In hypoxia group, the collagen cross-links in the lung tissue was significantly higher than that in normoxia group and hypercapnia group (P<0.01). Importantly, collagen cross-links in the lung tissue of hypoxia+hypercapnia group was significantly lower than that in hypoxia group (P<0.01). There was no significant difference in collagen cross-links between hypercapnia group and normoxia group. The expression of LOX at mRNA and protein levels and its activity in the pulmonary arteries of hypoxia group were significantly increased as compared with normoxia group (P<0.01). Furthermore, the expression of LOX at mRNA and protein levels and its activity in the pulmonary arteries in hypoxia+hypercapnia group were lower than those in hypoxia group (P<0.01). CONCLUSION: Hypoxia not only up-regulates LOX but also promotes collagen cross-linking in the rat lung, which contributes to the development of pulmonary hypertension. Hypercapnia inhibits hypoxia-induced LOX expression and collagen cross-linking, therefore impairing the progress in hypoxia-induced pulmonary hypertension.     

北青龙衣褐变机制的研究

北青龙衣在由鲜品到干品的贮藏期间会出现失水、颜色变深等褐变现象。研究该褐变过程发生的机制,为控制褐变、提高药材的稳定性提供试验依据。采用北青龙衣鲜果果皮,4℃低温贮藏,研究其褐变度(browning degree,BD)、多酚氧化酶(polyphenol oxidase,PPO)、苯丙氨酸解氨酶(phenylalanine ammonialyase,PAL)、过氧化物酶(peroxidase,POD)活性变化、丙二醛(malondialdehyde,MDA)、总多酚(total polyphenols,TP)、还原糖含量及维生素C(Vitamin C,VC)含量变化。试验数据显示果皮颜色变深、TP含量下降、PPO活性降低、PAL活性上升、SOD和POD活性先上升后下降、MDA含量升高、还原糖和VC含量下降。BD与PPO、SOD、POD活性和TP含量呈显著负相关,与PAL活性呈显著正相关,与VC和还原糖无显著相关性。北青龙衣贮藏过程中发生了酶促褐变和非酶褐变,且以酶促褐变为主,PPO在酶促褐变中可能起主要作用。     

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。     

Overexpression of StRbohA in Arabidopsis thaliana enhances defence responses against Verticillium dahliae

Plant NADPH oxidases are key regulators of plant–microbe interactions and reactive oxygen species (ROS) are essential to plant defences against pathogens. A significant part in the role played by ROS has been ascribed to plant respiratory burst oxidase homologs (RBOHs). In potato (Solanum tuberosum), where RBOHs were previously shown to be involved in wound-induced oxidative burst, we assessed their expression after inoculation with Verticillium dahliae Kleb. and showed that StRbohA was the only homolog to be differentially induced in potato in response to inoculation. In order to investigate the potential role of this gene in plant protection against wilt diseases, we used Agrobacterium-mediated transformation of Arabidopsis to assess the effects of its overexpression on plant responses to V. dahliae. After inoculation with this pathogen, the transformed Arabidopsis line overexpressing StRbohA showed lower disease severity (percent damaged leaf area and vascular discoloration) as compared to the wild type. It also had higher ROS production and more cell death caused by hydrogen peroxide (H₂O₂), compared to the wild type. Suberization of root cells was also more pronounced in the line overexpressing StRbohA, and supports a possible role for StRBOHA in plant resistance to V. dahliae. Together, these findings indicate that overexpressed StRbohA in Arabidopsis enhances the ROS-mediated defence mechanisms against V. dahliae and can be a potential tool to improve plant resistance to this and other soilborne pathogens that cause wilts in economically important crops.     

Molecular characterization of Ascaridia galli from Bangladesh and development of a PCR method for distinguishing A. galli from Heterakis spp.

We analyzed the nuclear ribosomal internal transcribed spacer (ITS) 1 and ITS2 sequences for Bangladesh isolates of Ascaridia galli, and we determined that the sequences were unreliable as molecular markers for distinguishing A. galli from other Ascaridia species, because the sequences showed high identity with that of A. columbae. However, the ITS1 sequences were available for designing PCR primers distinguishable between Ascaridia galli and Heterakis spp. Bangladesh isolates of A. galli constituted a monophyletic clade along with other geographical isolates in the cytochrome c oxidase subunit I (COI) phylogenetic tree, however, we could not clarify the phylogenetic relationships between A. galli and other Ascaridia spp., because their available sequences in GenBank were very few. The developed PCR method using DNA from A. galli and Heterakis spp. eggs would enable differential diagnosis of the individual infections in the future.     

红三叶多酚氧化酶特性的研究

红三叶(Trifolium pretense L.)多酚氧化酶仅作用于邻苯酚,与对苯酚、间苯酚和一元酚无作用,而且对不同的邻苯酚其活性也表现很大差异,其中以咖啡酸作底物时活性最大.红三叶多酚氧化酶反应的最适pH值为6.5～7.0,最适温度为25～30℃,50℃时热稳定性相对较好.     

葡萄糖氧化酶基因植物表达载体的构建

为进一步开展转基因研究创造条件,用限制性内切酶将葡萄糖氧化酶(GOD)基因从pMD/GO载体上切下,定向连接到植物表达载体pROKⅡ的CaMV35S启动子下游和NOS终止子上游,成功地构建了GOD基因植物表达载体pROK/GO。利用冻融法将此表达载体导入根癌农杆菌LBA4404中,提取转化质粒,经PCR和酶切鉴定表明,GOD基因植物表达双元载体构建成功。     

软儿梨冷冻贮藏过程酚类物质与多酚氧化酶的区域化分布

为研究冷冻贮藏中不同部位酚类物质和多酚氧化酶的分布特征,以青海省地方梨优良品种‘软儿梨’果实为研究材料,采用高效液相色谱法和比色法测定软儿梨果实后熟(室温)、冻藏(-18℃)和解冻(室温)3个时期的果皮、果肉中总多酚、绿原酸的含量和多酚氧化酶活性。结果表明:软儿梨果实后熟后,酚类物质果肉?果皮,多酚氧化酶则果皮?果肉;冷冻贮藏过程中,不同部位的酚类物质和多酚氧化酶均呈现不同程度的升高,但绿原酸在果皮急剧下降、果肉中略有增加。解冻后总体呈下降趋势,2个部位总酚、多酚氧化酶下降明显,而果皮中的绿原酸变化不大。研究表明:软儿梨果实后熟后,酚类物质和多酚氧化酶分布呈现区域化,冷冻贮藏加剧了酚类物质和多酚氧化酶活性的积累,导致了软儿梨冷冻贮藏后果皮先于果肉发生了褐变。