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。     

重组GSTA3蛋白对福美双诱导的肉鸡TD中抗凋亡基因BAG-3表达的影响

【目的】肉鸡胫骨软骨发育不良（TD）是肉鸡常见的一种骨骼性疾病,研究重组GSTA3蛋白对福美双诱导的TD肉鸡软骨细胞中抗凋亡基因BAG-3表达的影响,为治疗TD提供新的思路和方法。【方法】将120羽1周龄肉雏鸡随机分为6组（编号为A、B、C、D、E、F组）。A、B、C组为基础日粮对照组,D、E、F组为添加福美双日粮诱导TD组。试验饲喂福美双2 d诱发TD,在添加福美双第1、3、5、7天,腿部肌肉注射重组鸡GSTA3蛋白和磷酸盐缓冲液,A组与D组注射（100 μg·kg ^-1）磷酸盐缓冲液;B组与E组注射低剂量（100 μg·kg ^-1）GSTA3;C组与F组注射高剂量（200 μg·kg ^-1）GSTA3。试验历时23 d。添加福美双后1、2、4、6、10和15 d采集胫骨生长板。通过Real-time qPCR检测BAG-3基因的mRNA水平,利用免疫组化来检测BAG-3蛋白表达水平。【结果】Real-time qPCR结果显示,TD损伤修复期内,相比较于基础日粮对照组,福美双对照组肉鸡胫骨生长板中BAG-3 mRNA的表达水平基本都显著上调（P<0.05）;相比较于福美双对照组,E和F组在第2、4、10、15天都有显著差异,且在第10和15天显著低于福美双对照组（P<0.05）,表明与D组相比恢复较快。免疫组化结果表明BAG-3蛋白在肉鸡胫骨软骨细胞的增殖区和前肥大区无表达,只在肥大区细胞质中表达;福美双组与空白对照组相比,BAG-3蛋白表达增加;福美双高低剂量组与未注射蛋白的福美双组相比,重组GSTA3增加了肥大区的蛋白表达水平（第10和15天）。【结论】在福美双诱导肉鸡发生TD的过程中,GSTA3重组蛋白能够通过调控BAG-3表达参与凋亡途径,抑制细胞凋亡。在TD损伤修复期,注射GSTA3后使抗凋亡基因BAG-3蛋白表达增强,从而可参与细胞凋亡来缓解TD损伤,使得肉鸡TD生长板功能较快地恢复正常。     

蓖麻GST的基因克隆及生物信息学分析

谷胱甘肽S-转移酶(glutathione S-transferase,GST)是谷胱甘肽结合反应的关键酶,催化谷胱甘肽结合反应的起始步骤,主要存在于胞液中,有多种形式,其在病理学上有一定的重要性。本研究通过克隆GST基因以及对其进行相关的生物信息学分析。利用聚合酶链式反应(polymerase chain reaction,PCR)技术扩增该基因,经分析该基因可编码219个氨基酸,并且通过相关分析得到这219个氨基酸组成的蛋白为稳定的亲水性蛋白,有一个明显的区段存在信号肽,在该蛋白中α-螺旋和无规则卷曲是主要的二级结构方式,根据序列同源比对,蓖麻、麻疯树和榴莲聚类到一起,说明相对于其他物种,它们的同源性较高。这些分析结果为蓖麻谷胱甘肽S-转移酶生物合成及其功能的进一步研究提供一定的理论与实践依据。     

Effect of gold nanoparticles on reversing adriamycin resistance of human hepatocellular carcinoma HepG2/ADM cells

AIM: To investigate whether gold nanoparticles (GNPs) reverses adriamycin (ADM), resistance of human hepatocellular carcinoma drug-resistant cell line HepG2/ADM and to explore the potential mechanism. METHODS: The sensitivities of HepG2 cells and HepG2/ADM cells to ADM were tested by MTT assay before and after GNPs pretreatment. The apoptotic rate was examined by flow cytometry. The concentration of ADM in HepG2/ADM or HepG2 cells was determined by ultraviolet-visible spectrophotometer. The content of glutathione (GSH) in HepG2/ADM or HepG2 cells by DTNB method. RESULTS: The half maximal inhibitory concentrations (IC₅₀) of ADM for HepG2/ADM cells were(29.46±1.73) mg/L and (15.18±0.85) mg/L before and after GNPs pretreatment,respectively. The IC₅₀ of ADM for HepG2 cells was (9.16±2.03) mg/L before pretreatment. The apoptotic rate in GNPs+ADM group was higher than that in ADM group (P<0.05). The concentration of ADM in HepG2/ADM group was lower than that in HepG2 group (P<0.01). After GNPs pretreatment, the concentration of ADM in HepG2/ADM cells was higher than that before pretreatment. The content of GSH in HepG2/ADM group was higher than that in HepG2 group (P<0.01). After GNPs pretreatment, the content of GSH in the HepG2/ADM cells was lower than that before pretreatment. CONCLUSION: Gold nanoparticles can reverse ADM resistance of human hepatocellular carcinoma drug-resistant cell line HepG2/ADM, reduce the content of GSH and increase the concentration of ADM in HepG2/ADM cells.     

Schisandrin B protects against solar irradiation-induced oxidative injury in BJ human fibroblasts

The effects of schisandrin B (Sch B) and its analogs on solar irradiation-induced oxidative injury were examined in BJ human fibroblasts. Sch B and schisandrin C (Sch C) increased cellular reduced glutathione (GSH) level and protected against solar irradiation-induced oxidative injury. The photoprotection was paralleled by decreases in the elastases-type protease activity and matrix-metalloproteinases-1 expression in solar-irradiated fibroblasts. The cytochrome P-450-mediated metabolism of Sch B or Sch C caused ROS production. The results suggest that by virtue of its pro-oxidant action and the subsequent glutathione antioxidant response, Sch B or Sch C may offer the prospect of preventing skin photo-aging.     

Biochemical and histopathological studies to assess chronic toxicity of triazophos in blood, liver and brain tissue of rats

Triazophos, O,O-diethyl-1-H-1,2,4-triazol-3-yl phosphorothioate, (TZ) is an organophosphorus pesticide which is extensively used in agriculture for controlling insect pests. Except a FAO/WHO report no study has investigated its short-term toxicity with respect to its potential to cause biochemical and histopathological alterations. The present study was designed to identify the effect of TZ at different doses (1.64, 3.2 and 8.2 mg/kg) on the oxidative stress parameters in blood as well as organs involved in xenobiotic metabolism (liver and brain) following chronic exposure for 90 days. Moreover, the study also delineates the effect of TZ on the histo-architecture of these organs. The results indicated a dose dependent induction (p < 0.001) of oxidative stress, as evident by increased malondialdehyde (MDA) level and compromised antioxidant defense including glutathione S transferase (GST) activity, glutathione (GSH) content and ferric reducing ability of plasma (FRAP) in blood, and increased MDA level with concomitantly decreased GSH content in tissues, following chronic exposure to TZ. The ratio of MDA: FRAP in blood was found to be increased following chronic exposure to TZ and may serve as a suitable indicator of severity of oxidative damage. Onset of such biochemical alterations is one of the early adaptive responses to TZ exposure which leads to histopathological alterations in terms of diffuse fatty changes expanding from mid-zonal area to whole lobule in liver. However, increased oxidative stress did not bring any morphological alteration in brain. The present study concludes that induction of oxidative stress, leading to subsequent histopathological alterations in liver, is an important mechanism underlying the TZ induced chronic toxicity.     

氨氮胁迫对四鼻须鲤鱼肝脏GST酶活力的影响

实验采用生态毒理学方法,以NH4Cl为实验药物,设置0、50、150和250mg/L实验浓度。通过对野生和养殖群体的四鼻须鲤鱼氨氮对肝脏谷胱甘肽巯基转移酶（GST）活力的影响,比较两个群体抗氨氮能力。结果表明：在相同的氨氮浓度和胁迫时间下,四鼻须鲤鱼养殖群体GST酶活力的变化趋势与野生群体大致相同。除250mg/L浓度组在胁迫5d和10d时,野生群体酶活性略低于养殖群体外,在其余时间点的野生群体酶活性均高于养殖群体,而且,在高浓度氨氮胁迫下,野生群体肝脏的GST酶活性达到峰值的时间比养殖群体更短,显示出较强的抗氨氮能力。     

Glutathione, Cysteine, and Ascorbate Concentrations in Clinically Ill Dogs and Cats

Background: Oxidative stress plays a role in the pathogenesis of many systemic diseases. Hospitalized human patients are glutathione, cysteine, and ascorbate deficient, and antioxidant depletion has been correlated with poor clinical outcome. To date little is known about antioxidant concentrations in hospitalized veterinary patients. The purpose of this study was to determine whether ascorbate, cysteine, or glutathione depletion is present in ill dogs and cats compared with healthy controls.
Hypothesis: Clinically ill dogs and cats would be antioxidant depleted, and depletion would correlate with illness severity and clinical outcome.
Animals: Clinically ill client-owned dogs (n = 61) and cats (n = 37), healthy control dogs (n = 37) and cats (n = 33).
Methods: Prospective, observational, case control study. Erythrocyte reduced glutathione, plasma cysteine, and plasma ascorbate were quantified using high-performance liquid chromatography.
Results: Clinically ill dogs had significantly lower erythrocyte glutathione concentrations (1.22 mM, range 0.55–3.61) compared with controls (1.91 mM, range 0.87–3.51; P = .0004), and glutathione depletion correlated with both illness severity ( P = .038) and mortality ( P = .010). Cats had higher ascorbate concentrations when ill (10.65 μM, range 1.13–25.26) compared with controls (3.68 μM, range 0.36–13.57; P = .0009).
Conclusions and Clinical Importance: Clinically ill dogs had decreased erythrocyte glutathione concentrations, which could be a marker of illness severity and prognostic of a poor outcome. Clinically ill cats had an unexpectedly high plasma ascorbate, which could represent a unique species response to oxidative stress.     

Ontogeny of glutathione and glutathione-related antioxidant enzymes in rat liver

We conducted a comprehensive characterization of the ontogeny of glutathione (GSH) and its related enzymes in rat liver. GSH content and activities of glutathione reductase (GR), cytosolic glutathione-S-transferases (GST), and glutathione peroxidase (GPx) were determined in male rat livers (n = 4) at different developmental stages. Our results indicate total hepatic GSH content and GR, GST, and GPx activity were low in the perinatal period. GSH content remained relatively constant throughout postnatal development, but GR, GST, and GPx activity underwent significant increases to attain adult levels by late post-weaning. Whether the ontogenic pattern of GSH and its related enzymes explain, in part, the altered susceptibility of neonates to some toxicants requires further investigation. Our study provides fundamental biological information on the ontogeny of cytosolic antioxidant/detoxification enzymes in a relevant toxicological risk assessment species. Additional studies are needed to fully characterize the ontogeny of other xenobiotic detoxification enzymes to further promote the rat as a developmental toxicology model that can identify toxicokinetic reasons for differences in susceptibility to toxicity between the neonate and adult.     

SO2对家蚕血淋巴中谷胱甘肽含量及相关酶活性的影响

通过分析在SO2胁迫下家蚕5龄幼虫血淋巴中谷胱甘肽(GSH)含量及相关酶活性的变化,探讨SO2对家蚕的毒理作用机制。当对家蚕5龄幼虫每天以40 mg/m3SO2熏气刺激6 h,蚕体血淋巴中的GSH含量显著降低,雄蚕GSH的平均含量为49.8μmol/L,雌蚕为41.0μmol/L,分别为对照的73.80%和55.76%,而谷胱甘肽硫-转移酶(GST)活性显著升高,雄蚕GST的平均活性为31.86 U/L,雌蚕为31.80 U/L,分别高于对照46.15%和27.30%。与此同时,γ-谷氨酰转肽酶(γ-GT)活性在蚕体中也有不同程度的升高,其中在雄蚕体内的变化极为显著;谷胱甘肽过氧化酶(GSH-Px)和谷胱甘肽合成酶系的活性在蚕体内均无显著变化。以上结果表明:SO2刺激可对家蚕血淋巴中的GSH含量及其相关酶的活性产生影响,且雌、雄蚕对SO2的敏感性不同;GSH含量的减少及其相关酶活性的上升,说明SO2对蚕体的毒害作用与其降低蚕体抗氧化物质水平、削弱抗氧化防御系统有关。