Progress in the research of GSH in cells

Glutathione (GSH), γ-Glu-Cys-Gly, is one of the most abundant small non-protein thiol molecules in mammalian tissues, particu- larly in the liver. Although glutathione is present in thiol-reduced (GSH) and disulfide oxidized (GSSG) forms, the predominant form is GSH and its content can exceed 10 mmol/L in liver cells. As an important intracellular reductant, GSH has many biological functions in cells. Its major function is as an anti-oxidant as it can protect proteins from oxidation by reversible posttranslational modification (glutathionylation) and decrease reactive oxygen species-mediated damage. However, it does have numerous other functions, including to chelate metal irons; enhance the absorption of iron, selenium and calcium; participate in lipid and insulin metabolism; regulate cellular events such as gene expression, DNA and protein synthesis, cell proliferation and apoptosis, redox-dependent signal transduction pathways, cytokine production and the immune response; and control protein glutathionylation. Therefore, GSH plays important roles in cell survival and health, and an imbalance in the GSH level can lead to many diseases. In this review, we provide an overview of the function of GSH in mammalian cells and discuss future research of GSH.     

胃癌高发区不同人群血浆总抗氧化力及谷脱甘肽过氧化物酶活性的研究

用微弱化学发光法和酶联分光光度技术研究胃癌高发区－－甘肃省武威地区不同人血浆总抗氧化力（TRAC）和谷胱甘肽过氧化物酶（GSH－Px）活性，结果表明：中度表浅性胃炎18例、21例明显降低(p-≤0．01），但各组间血浆GSH－Px活性无显著性差异（P＞0．05），提示当地胃癌高发可能与血产供销 TRAC水平的降低有关，而与GSH－Px活性改变无明显相关。     

The effects of vitamin A nutritional status on glutathione levels and microsomal lipid peroxidation in rat lung

Summary In vitamin A-deficient rats, the glutathione level in lung was diminished and microsomal lipid peroxidation much increased. In vitamin A-loaded animals, however, both were depressed below control. Thus vitamin A protection against lipid peroxidation is independent of glutathione.Acknowledgments. We thank Dr L. Y. Y. Fong and Mr David Y. H. Woo for preparing the animals used in this research, for retinol determinations and for valuable discussions, and also the China Medical Board of New York and the University of Hong Kong for the award of a Fellowship to V.P.     

活性氧对人体中与衰老相关物质的影响

研究了过硫酸按-TEMED体系产生的超氧负离子对健康人血液中几种生物活性物质的作用，结果表明，可以使细胞膜发生脂质过氧化作用，产生丙二醒（MDA），使生物膜造成损伤．同时使血液中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH－PX）活性均有所降低．通过对CAT纯品的研究揭示出在血液中经SOD，的作用产生毒性更强的·OH，它是使生物膜破坏及各种酶活性降低的主要原因．     

用化学发光法研究谷胱甘肽对羟自由基氧化鸟嘌呤的保护作用

利用鸟嘌呤-过氧化氢-钴（Ⅱ）化学发光体系，结合流动注射技术研究了生物活性物质谷胱甘肽（GSH）对羟自由基氧化鸟嘌呤的保护作用．实验结果表明，GSH明显抑制鸟嘌呤的化学发光，且抑制率与GSH浓度成量效关系，达到50％抑制率的GSH浓度为6．5mg／L．该方法为进一步研究DNA的抗氧化损伤机制提供了有效的研究手段．     

Effects of dietary nickel on detoxification enzyme activities in the midgut of <Emphasis Type="Italic">Spodoptera litura</Emphasis> Fabricius larvae

Nickel accumulated in midugt of Spodoptera litura Fabricius could induce the expression of metallothionein, one of the most important detoxification proteins in organisms. In the present study, the effects of dietary nickel on the activities of detoxification enzymes, such as carboxylesterase （CarE） and giutathione S-transferase （GST） in the midgut of S. litura larvae have been studied to get an understanding of the detoxification mechanisms of S. litura larvae to excessive nickel. Results showed that CarE activities in the midgut of the 5th instar larvae decreased at lower levels of nickel （≤5 mg/kg）, while increased with increasing nickel doses at higher levels of nickel （≥10 mg/kg） exposure in successive 3 generations. CarE activities of the 6th instar larvae were also characterized as inhibited at low levels of nickel exposure, and improved at higher levels in the 1st generation. CarE activities of 6th instar larvae in the 2nd and 3rd generations were all lower than that in control. However, GST activities in the midgut of the 5th and 6th instar larvae all increased with increasing nickel doses （1 --20 mg/kg） in diets.     

N末端缺失萝卜谷胱甘肽磷脂氢过氧化物酶的表达、纯化及结构预测

将N末端定位信号缺失萝卜PHGPx（RsPHGPx）的cDNA 片段克隆到表达载体pGEX-6P-1上, 并转化至大肠杆菌内进行表达. 通过GST亲和层析、离子交换层析和凝胶过滤层析, 制备了用于晶体学研究的RsPHGPx. 其浓度约为10 g/L, 纯度超过95％, 具有PHGPx活性. 三维结构同源建模显示RsPHGPx的结构为典型的硫氧还蛋白折叠形式.     

促肝细胞生长素联合还原型谷胱甘肽治疗重度慢性乙型肝炎

目的观察促肝细胞生长素联合还原型谷胱甘肽治疗重度慢性乙型肝炎的临床疗效。方法选择60例重度慢性乙型肝炎随机分为对照组和治疗组各30例,比较治疗前后两组病人的临床症状和生化指标等方面的变化。对照组给予甘草酸二铵、香丹等抗炎保肝支持及防治并发症治疗;治疗组在此基础上加用促肝细胞生长素和还原型谷胱甘肽联合治疗。结果治疗组疗效明显优于对照组,治疗组在临床症状和生化指标改善等方面与对照组比疗效差异有显著性意义（P〈0．05）。结论两药联用有助于改善临床症状和生化指标。缩短重度慢性乙型肝炎的病程,防治疾病进展。     

伽马辐照对斑马鱼胚胎谷胱甘肽还原酶活性及基因表达的影响

谷胱甘肽还原酶(GR)是一种重要的抗氧化酶,其在生物体的抗氧化系统中起着关键作用.为了研究伽马辐照对斑马鱼胚胎GR活性及基因表达的影响,本实验用不同的辐照剂量(0.01、0.05、0.1、0.5和1 Gy)分别对三个发育时期(原肠胚期、体节期和咽囊期)的斑马鱼胚胎进行了辐照处理.分析了各组GR的活性,并采用荧光实时定量PCR技术(qRT-PCR)检测了GR基因的表达水平.结果显示,伽马辐照以剂量依赖的方式调整了斑马鱼胚胎GR的活性及GR基因的表达.随着辐照剂量的增加,GR活性及其mRNA的相对表达量呈现先升高后回落的趋势.这表明,GR是一种可以对伽马辐照作出早期预警的生物标志物.斑马鱼胚胎可以通过上调GR基因的表达,来增加GR的合成,从而提高对伽马射线的防御能力.但是,这种反应和调整是有限度的.咽囊期胚胎比其它两个时期的胚胎对伽马射线的防御能力更强.     

Biosynthesis of selenosubtilisin: A novel way to target selenium into the active site of subtilisin

Glutathione peroxidase （GPx, EC1.11.1.9）, an important anti-oxidative selenoenzyme, can catalyze the reduction of harmful hydroperoxides with concomitant glutathione, thereby protecting cells and other biological issues against oxidative damage. It captures considerable interest in redesign of its function for either the mechanism study or the pharmacological development as an antioxidant. In order to develop a general strategy for specifically targeting and operating selenium in active sites of enzymes, the catalytically essential residue selenocysteine （Sec） was first successfully bioincorporated into the catalytic center of subtilisin by using an auxotrophic expression system. The studies of the catalytic activity and the steady-state kinetics demonstrated that selenosubtilisin is an excellent GPx-like biocatalyst. In comparison with the chemically modified method, biosynthesis exhibits obvious advantages： Sec could be site-directly incorporated into active sites of enzymes to overcome the non-specificity generated by chemical modification. This study provides an important strategy for specifically targeting and operating selenium in the active site of an enzyme.