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.     

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。     

Dough rheological properties and noodle-making performance of non-waxy and waxy whole-wheat flour blends

Dough rheological properties and noodle-making performance of non-waxy whole-wheat flour (WWF) with partial- or full-waxy (PW- or FW-) WWF substitution were studied. The substitution levels were 0, 250, 500, 750, and 1000 g/kg, respectively. FW-WWF reduced the peak viscosity and pasting temperature of WWF blends as its substitution level was increased due to its higher proportions of B-type starch granules and short amylopectin chains, while PW-WWF increased peak viscosity with the increasing substitution level because of its higher amylopectin content. As demonstrated by farinograph and rheometer measurements, FW-WWF interfered with gluten development because of the increased competition for water by arabinoxylans and amylopectin; however, PW-WWF enhanced dough strength due primarily to its increased protein content. Consequently, FW-WWF showed a detrimental effect on cooked noodle texture as the cooked noodle hardness was reduced by 50% at the 1000 g/kg substitution level. In contrast, PW-WWF enhanced noodle integrity and elasticity by increasing cooked noodle cohesiveness and resilience by 10.1% and 14.8%, respectively, at the 1000 g/kg substitution level. The results suggest that with waxy WWF substitution, the changes in starch composition, arabinoxylans, and protein content could modify the interactions among flour components and influence the quality characteristics of noodle products.     

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。     

枇杷果实发育过程中山梨醇含量及相关酶活性的变化

以果型和品质差异较大的黄肉枇杷(Eriobotrya japonica Lindl.)‘解放钟’和白肉枇杷‘白梨’的果实为试材,测定果实发育过程中山梨醇、果糖、葡萄糖含量及与山梨醇代谢相关酶--山梨醇脱氢酶(Sorbitol dehydrogenase,SDH)和山梨醇氧化酶(Sorbitol oxidase,SOX)活性的变化,并探讨山梨醇代谢与枇杷果实生长发育和品质形成的关系。结果表明:2个枇杷品种果实中山梨醇含量均随果实发育进程呈下降趋势,且‘白梨’枇杷果实中山梨醇含量高于‘解放钟’。在花后35~80 d,2个枇杷品种幼果中葡萄糖和果糖含量均较低且相对稳定,而花后95~140 d果实中葡萄糖和果糖含量逐渐上升,并在花后125~140 d果实成熟期急剧增加而达到峰值;果实中葡萄糖含量在2个品种间差异不显著(p0.05),花后125~140 d的果实成熟期,‘白梨’果实中果糖含量显著高于‘解放钟’(p0.05)。果实发育过程中SDH和SOX的活性均呈先降后升的变化趋势。说明2个枇杷品种果实发育过程中山梨醇、果糖、葡萄糖含量及SDH和山SOX的活性表现出相似的变化规律,但果实山梨醇含量与SDH和SOX的活性在果实膨大期均存在显著差异(p0.05),这可能是黄肉枇杷与白肉枇杷品质差异的关键。     

铜锌对华北落叶松多酚氧化酶活性的影响

以华北落叶松3年生幼苗为试验材料,对不同生育期4种处理(对照、锌、铜、铜 锌)下各器官(叶、枝、干、主根、侧根)多酚氧化酶(PPO)活性进行测定分析,结果表明:华北落叶松幼苗各器官内多酚氧化酶活性差异极为显著,干中PPO活性在3月达225.2 U·g-1,而叶中PPO活性在5月仅为57.3 U·g-1.叶子中PPO活性呈递增变化趋势,10月最高,枝、干、主根、侧根的PPO活性在年循环周期内呈规律性变化,7月均达高峰期.铜、锌对各器官内PPO活性有明显的影响,而且与生育期有关.铜对PPO活性的增强作用达显著水平,尤以枝最为突出,10月份铜处理后枝中PPO活性比对照提高95.1%;锌对主根PPO活性的提高也有较明显影响,对其他器官PPO活性有不同程度的抑制作用.10月份锌处理后干中PPO活性比对照下降35.5%;铜、锌互作对干和主根PPO活性的提高表现出较大的协同作用.华北落叶松幼苗体内铜元素含量与PPO活性呈显著的正相关,铜、锌元素通过影响PPO活性而影响幼苗的生长及抗寒性.     

中国药用植物中单宁及多酚类化学成分的研究

从１２种中国药用植物中分离到１１３个单宁及多酚类化合物，经波谱分析及化学方法鉴定了它们的结构。其中１９个为新化合物。     

桉木抽出物中多酚化合物的特性研究

采用化学分析和光谱分析相结合的方法，从制浆造纸的角度，重点对雷林一号桉木的多酚化合物的特性进行了研究。结果表明：幼龄材的抽出物含量较少，适宜于制浆的采伐期为５～７年。１５年生雷林一号桉木心材中的多酚类化合物以水解单宁为主（占原料的５．９８％），其次为缩合单宁（１．９１％）。稀碱预处理桉木，可除去大部分多酚类化合物。存放过久的桉木中多酚化合物大部分变成‘黄粉’，既不溶于水也不溶于丙酮，只溶于稀碱或亚硫酸钠。桉木制浆前采用碱预处理可减轻多酚化合物对制浆产生的不良影响。     

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。