Heme oxygenase-1 protects renal function in septic rats via influencing expression of thrombomodulin

AIM: To investigate the protective effect of heme oxygenase-1 (HO-1) on the kidney of septic rats and the influence of HO-1 on the expression of thrombomodulin (TM) in the kidney. METHODS: Sepsis in rats was developed with cecal ligation and puncture (CLP). The septic rats were randomly divided into sham group, CLP group, CLP+HO-1 inducer group and CLP+HO-1 inhibitor group (n=18). The plasma levels of creatinine (Cr), cystatin-C (Cys-C), carboxyhemoglobin (COHb), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and TM, and the changes of prothrombin time (PT) and activated partial thromboplastin time (APTT) in each group were measured. Histopathological examination was performed in the kidney. The expression of TM in the kidney tissue was detected by Western blot. RESULTS: Compared with sham group, significantly elevated plasma levels of Cr, Cys-C, TNF-α, IL-1β and TM (P<0.05), shortened PT and APTT (P<0.05), significantly increased microthrombus formation, and lowered TM expression in the kidney (P<0.05) of CLP group were observed. The administration of hemin lowered the plasma levels of Cr, Cys-C, TNF-α, IL-1β and TM (P<0.05), prolonged PT and APTT (P<0.05), attenuated microthrombus formation, and up-regulated the expression of TM in the kidney (P<0.05). In contrast, ZnPP had the opposite effects. CONCLUSION: HO-1 increases the expression of TM in the kidney and exerts anticoagulatory and antiinflammatory effects, thereby improving renal function in the septic rats.     

茶树叶绿素合成相关基因克隆及在白叶1号不同白化阶段的表达

高等植物叶绿素的生物合成对其正常光合作用起关键作用。本文根据前期芯片杂交结果, 采用RT-PCR和RACE技术克隆了3个茶树叶绿素合成相关基因, 分别为谷氨酸-tRNA还原酶(CsGluTR)、叶绿素合酶(CsChlS)、叶绿素酸醋氧化酶(CsCAO), 对应的GenBank的登录号为HQ660371、HQ660370、HQ660369。序列分析表明, CsGluTR基因全长2165 bp, 开放阅读框长1665 bp, 编码554个氨基酸, 推测的蛋白分子量约为60.6 kD, 理论等电点为8.78;CsChlS基因全长1463 bp, 其中开放阅读框长1125 bp, 编码374个氨基酸, 推测的蛋白分子量约为40.5 kD, 理论等电点为8.58;CsCAO基因全长2146 bp, 其中开放阅读框长1611 bp, 编码536个氨基酸, 推测的蛋白分子量约为60.8 kD, 理论等电点为8.03。比对分析表明, 3个基因编码的氨基酸序列与其他植物中同源基因的相似性均在70%以上。利用荧光定量PCR技术检测3个基因在不同白化阶段的表达,表明, CsChlS和CsCAO基因具有明显的表达协同性, 它们在叶片中的表达量与叶片的颜色变化高度同步;而CsGluTR在白化叶片和正常叶片中的表达差异相对较小, 同时在新生芽叶转绿过程中最先恢复正常表达水平。说明在白化叶片中, 叶绿素的合成机制受到较大影响, 叶绿素合成受阻导致的叶片内色素类物质含量降低或消失是叶片白化的直接原因。     

Senescence of Top Three Leaves in Field‐Grown Rice Plants

Abstract

The top three leaves play important roles in biomass production and grain yield of rice (Oryza sativa L.) crop since the three leaves not only assimilate majority of carbon for grain filling during ripening phase, but also provide large proportion of remobilized‐nitrogen (N) for grain development during their senescence. The objectives of this study were to (a) compare senescence of the top three leaves and (b) compare the changes in N, chlorophyll, and ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) contents of the top three leaves after their full expansion in field‐grown rice plants. When the basis of comparison among the top three leaves was plant age in terms of days after transplanting (DAT), senescence generally started earliest in ?3rd leaf, intermediate in ?2nd leaf, and latest in flag leaf. If the basis of comparison among the top three leaves was leaf age in terms of days after full leaf expansion (DAFE), it was not clear which leaf senesced earlier. Senescence rate was generally greatest in flag leaf, intermediate in ?2nd leaf, and smallest in ?3rd leaf. Ribulose‐1,5‐bisphosphate carboxylase/oxygenase content declined earlier, and at a faster rate than N and chlorophyll contents during the senescence of all top three leaves. Correlation analysis indicated a close relationship between N and chlorophyll contents. Ribulose‐1,5‐bisphosphate carboxylase/oxygenase content correlated with N content better than with chlorophyll content. The suitability of N, chlorophyll, and Rubisco contents for quantifying the leaf senescence of field‐grown rice plants is discussed.     

Responses of leaf photosynthesis and plant growth to altered source–sink balance in a Japanese and a Dutch tomato cultivar

We investigated the effects of altered source/sink ratio by leaf or fruit pruning on leaf photosynthetic characteristics and whole-plant growth of ‘Momotaro York’, a Japanese cultivar, and ‘Dundee’, a Dutch cultivar and verified a hypothesis for sink-limitation of plant growth proposed by Tanaka and Fujita (1974). Plants were grown hydroponically with a high-wire system in a greenhouse for 11 weeks. Light-saturated photosynthesis of young, fully expanded leaves measured at atmospheric CO₂ partial pressure of 37 Pa and at an intercellular CO₂ partial pressure of 20 Pa was not influenced by alteration of source/sink ratio for either cultivar. Although soluble sugars were accumulated in leaves under high source/sink conditions, the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase, a rate-limiting factor for CO₂-limited photosynthesis, was not affected by the altered source–sink balance. Net assimilation rate of a whole plant increased with decreasing source/sink ratio, but this can be accounted for by the changes in light interception per unit leaf area, without taking the source–sink relationship into consideration. It was concluded that the altered source/sink ratio did not change leaf photosynthetic capacity and the sink-limitation hypothesis cannot be applied to either cultivar under the conditions of the present study.     

Induction of heme oxygenase-1 by Macleaya cordata extract and its constituent sanguinarine in RAW264.7 cells

Macleaya cordata (plume poppy) is used in traditional Chinese medicine for its anti-inflammatory and antibacterial activities. In this study, we examined whether M. cordata extract and/or its major alkaloid constituents, protopine, allocryptopine, sanguinarine and chelerythrine activate the Nrf2 signalling pathway which regulates the expression of cytoprotective enzymes including heme oxygenase-1 (HO-1) and thioredoxin 1. In murine macrophage RAW264.7 cells, M. cordata extract increased both mRNA and protein levels of HO-1. Of the alkaloids examined, only sanguinarine appeared to be responsible for these effects. At the concentration of 2 μM, sanguinarine induced nuclear accumulation of Nrf2, increased the expression of Hmox1 gene encoding HO-1 and elevated HO-1 protein levels. Sanguinarine-induced Hmox1 mRNA expression was suppressed by SB203580, a pharmacologic inhibitor of p38 mitogen-activated protein kinases (p38 MAPKs). The upregulation of HO-1 in RAW264.7 cells by sanguinarine was, however, accompanied by decrease in cell viability. Nonetheless, sanguinarine at micromolar, non-cytotoxic concentrations elevated protein levels of HO-1 and thioredoxin 1 in primary cultures of human hepatocytes. We conclude that sanguinarine may, under appropriate conditions, increase the capacity of the enzymatic antioxidant defence system via activation of the p38 MAPK/Nrf2 pathway.     

Effect of the β‐carotene oxygenase 2 genotype on the content of carotenoids,retinol and α‐tocopherol in the liver,fat and milk of rabbit does,reproduction parameters and kitten growth

Mutations in the β‐carotene oxygenase 2 (BCO2) gene can impair the function of the enzyme that breaks down carotenoids. As a result, gradual accumulation of unoxidized carotenoids in animal tissues gives them a yellow colour. The aim of the study was to determine the content of carotenoids, retinol and α‐tocopherol in the liver, fat and milk of rabbit does with three different genotypes determined by AAT‐deletion mutation at codon 248 of the BCO2 gene and to find out whether differences in the concentrations of the above compounds in the tissues and milk of the does affect reproduction parameters and the rearing rate of kittens. The experimental materials comprised 36 does, 12 of each genotype of the BCO2 gene, with their litters. Females with their litters were placed in individual cages, on deep litter. Between days 7 and 13 of lactation, samples of milk were collected from the does. The kittens stayed with their mothers until 35 days of age. After weaning, the does were sacrificed. Tissue samples of liver and perirenal fat were collected for chemical analyses. Additionally, based on samples taken from one female, RNA expression levels were determined from the mammary gland and liver, adipose tissue and skin. It was found that homozygous does with deletion at codon 248 of the BCO2 gene were characterized by considerably higher concentrations of xanthophylls and beta‐carotene in the liver, adipose tissue and milk than does with the remaining genotypes. However, the differences in the content of the above compounds in milk had no influence on litter weight or the number and rearing rate of kittens. Additionally, RNA expression of the BCO2 gene was found in the mammary tissue of lactating doe and its level was similar to those noted in the liver and adipose tissue.     

黄瓜幼苗Rubisco与Rubisco活化酶对光强的响应

以津优3号幼苗为试材,研究弱光(LL:100 μmol· m-2·s-1)、亚弱光(SL:300 μmol·m-2·s-1)下黄瓜幼苗光合速率(Pn)、核酮糖-1,5-二磷酸羧化/加氧酶(Rubisco)、Rubisco活化酶(RCA)活性及其基因表达量的变化.结果表明,11d弱光处理后,黄瓜幼苗叶片的Pn、Rubis...     

适度水分胁迫提高黄瓜幼苗光合作用弱光适应性

以黄瓜（Cucumis sativus L.）‘戴多星’品种为试材,在人工气候室内研究弱光（75 ~ 85 μmol · m-2 · s-1）下水分胁迫（40%基质最大持水量）对幼苗叶片光合、荧光特性及Rubisco含量的影响。结果表明,弱光逆境下,经20 d水分胁迫后的幼苗较正常水分条件下叶片光合色素（以单位面积计）含量提高,叶绿素a/b（Chl.a/b）下降,植株光饱和光合速率Asat、表观量子效率（AQY）、羧化效率（CE）、RuBP最大再生速率及光饱和点（LSP）均显著增加,CO2补偿点（CCP）显著下降,并且光系统Ⅱ（PSⅡ）最大光化学效率（Fv/Fm）、潜在光化学效率（Fv/Fo）、PSⅡ光化学量子效率（ΦPSⅡ）、光化学反应速率（Prate）、非环式电子传递速率（J）以及PSⅡ吸收光能向光化学反应分配比例P等叶绿素荧光参数呈增加趋势,PSⅠ和PSⅡ间激发能分配不平衡性（β / α–1）减小,叶片可溶性蛋白质含量和Rubisco大、小亚基含量上升,表明适度的水分胁迫有利于提高弱光下黄瓜幼苗的光合适应性;而正常光照下（500 ~ 600 μmol · m-2 · s-1）经水分胁迫后的黄瓜幼苗上述主要光合特性指标如Asat、RuBP最大再生速率、CE、ΦPSⅡ、Prate、J和P等有所下降,光系统间激发能分配不平衡性（β / α–1）增加,光合作用受到一定抑制。