藻类硒蛋白研究进展

硒是人体生长必须的基本元素。硒蛋白是由硒代半胱氨酸插入特异结合的蛋白。它在动物中的重要作用具有广泛的研究,而高等植物和酵母只有少量的相关研究。通过对藻类富硒机理、硒代谢途径、硒分布和形态、藻类硒蛋白等方面内容的总结、概括,重点介绍了藻类几种重要的硒蛋白。     

Variation in Vitamin E,Glutathione Peroxidase and Retinol Concentrations in Blood Plasma of Primiparous Sows and their Piglets,and in Vitamin E,Selenium and Retinol Contents in Sows' Milk

The concentrations of vitamin E, glutathione peroxidase (GSH-Px) and retinol were analysed in blood plasma of six primiparous sows during gestation and lactation and in their suckling piglets, and the concentrations of vitamin E, selenium and retinol were analysed in colostrum and in sows' milk. During late gestation the concentrations of vitamin E and retinol in blood plasma increased slowly while no changes were found for GSH-Px. This was followed by a large decline on the day of farrowing, when the lowest mean values of 1.07 mg, 0.15 mg and 2879 U per litre (l) of blood plasma, respectively, were recorded. During lactation the concentrations of vitamin E, GSH-Px and retinol in blood plasma of the sows increased to levels observed during late gestation. The highest mean concentrations of vitamin E (17.9 mg kg- 1 ), selenium (200 w g l- 1 ) and retinol (2.45 mg kg- 1 ) were found in colostrum, but with a large variation between sows. One week postpartum the concentrations of vitamin E, selenium and retinol in sow milk had decreased, on average by 86, 68 and 71%, respectively, after which only small changes occurred. Mean concentrations of vitamin E (1.77 mg l- 1 ), GSH-Px (682 U l- 1 ) and retinol (0.07 mg l- 1 ) were low in the blood plasma of the piglets at birth, but a large increase was observed on the next sampling occasion at the age of 3 days (vitamin E: 5.31 mg l- 1 ; GSH-Px: 996 U l- 1 ; retinol: 0.19 mg l- 1 ). The mean concentration of vitamin E in blood plasma of the piglets declined thereafter until the age of 6 weeks (1.04 mg l- 1 ), when there was a small increase, while the mean concentrations of GSH-Px and retinol continued to increase during the suckling period. A high correlation was found between the concentration of vitamin E in blood plasma and sidefat thickness of the sows (r = 0.94), and between the concentration of vitamin E in colostrum and vitamin E in blood plasma of the sows on the day of farrowing (r =0.77). High correlations were also found between the concentration of vitamin E in the blood plasma of the piglets at 3 days and 5 weeks of age and the concentration of vitamin E in colostrum (r = 0.78) and (r =0.89), respectively. The results indicate that vitamin E storage in the adipose tissues of the sow has a great influence on the supply of vitamin E via colostrum and milk to the piglet.     

Effect of glutathione on gelatinization and retrogradation of wheat flour and starch

Reduced glutathione (GSH) commonly exists in wheat flour and has remarkable influence on gluten properties. In this study, effect of GSH on the gelatinization and retrogradation of wheat flour and wheat starch were investigated to better understand the GSH-gluten-starch interactions in wheat flour. Compared with wheat starch, wheat flour showed significant decreases in peak and final viscosity, and gelatinization onset temperature with increasing GSH concentration. GSH depolymerized gluten and thereby broke down the protein barrier around starch granules to make the starch easily gelatinized. However, the interaction between GSH and wheat starch restrained starch swelling. GSH addition resulted in weakened structure with higher water mobility in freshly gelatinized wheat flour dispersions but decreased water mobility in wheat starch dispersions. After storage at 4 °C for 7 d, GSH increased elasticity and retrogradation degree in wheat flour dispersions but retarded retrogradation in wheat starch dispersions. The results indicated that GSH promoted retrogradation of wheat flour, which mainly attributed to the depolymerized gluten embedding in the leached starch chains, and inhibiting the re-association of amylose, and subsequently promoted the starch intermolecular associations and starch retrogradation. This study could provide valuable information for the control of the quality of wheat flour-based products.     

Characterization of acephate resistance in the diamondback moth Plutella xylostella

Decreased acetylcholinesterase (AChE) sensitivity and metabolic detoxification mediated by glutathione S-transferases (GSTs) were examined for their involvement in resistance to acephate in the diamondback moth, Plutella xylostella. The resistant strain showed 47.5-fold higher acephate resistance than the susceptible strain had. However, the resistant strain was only 2.3-fold more resistant to prothiofos than the susceptible strain. The resistant strain included insects having the A298S and G324A mutations in AChE1, which are reportedly involved in prothiofos resistance in P. xylostella, showing reduced AChE sensitivity to inhibition by methamidophos, suggesting that decreased AChE1 sensitivity is one factor conferring acephate resistance. However, allele frequencies at both mutation sites in the resistant strain were low (only 26%). These results suggest that other factors such as GSTs are involved in acephate resistance. Expression of GST genes available in P. xylostella to date was examined using the resistant and susceptible strains, revealing no significant correlation between the expression and resistance levels.     

以色列二穗短柄草谷胱甘肽过氧化物酶活性分析

二穗短柄草是一种理想的研究温带禾谷类植物和牧草的模式植物.本文对来源于以色列4个不同地区105个生态型二穗短柄草的根、茎、叶中的谷胱甘肽过氧化物酶(GSH-Px)活性进行了测定分析.结果表明:在贵州的自然气候条件下,以色列二穗短柄草根、茎、叶部位的GSH-Px活性存在显著性差异,其活性的高低顺序为:叶>茎>根;不同生态...     

Prophylactic effect of green tea polyphenols against liver and kidney injury induced by fenitrothion insecticide

The ameliorative effect of daily administrated dose of green tea extract (60 mg polyphenols/animal/day) was investigated on albino rats Rattus norvegicus (150-180 gm) intoxicated with 1/30 and 1/60 LD₅₀ fenitrothion organophosphate insecticide for 28 days. Blood samples were taken at 14 and 28 days for further biochemical parameters. Histopathological studies were carried out in the liver and kidney at the end of the experiment. Significant inhibition in plasma cholinesterase (ChE), a biomarker of Ops, was recorded. Damage in the liver and kidney tissues was observed and confirmed with elevation of plasma alanine aminotransferase (ALT), aspartate aminotaransferase (AST), albumin, urea and creatinine, as well as an elevation in the oxidative stress (OS) marker malondialdehyde (MDA). Decrease in total glutathione (GSH) content in erythrocytes and fluctuation in glutathione S-transferase (GST) activity in plasma was also observed. Green tea supplementation (60 mg/animal/day) partially counteracts the toxic effect of fenitrothion on oxidative stress parameters and repairs tissue damage in the liver and kidney, especially when supplemented to 1/60 LD₅₀ intoxicated animals depending on the duration. It seems that enzyme and metabolite markers of these organs need more time to be restored to the control level.     

舞毒蛾LdGSTe1基因的克隆及表达

从舞毒蛾(Lymantria dispar)3龄幼虫转录组文库中鉴定获得舞毒蛾谷胱甘肽S–转移酶基因全长c DNA,命名为Ld GSTe1。该基因全长651 bp,编码216个氨基酸。序列分析显示,Ld GSTe1蛋白氨基酸序列含有GST_C_Delta_Epsilon与GST_N_Delta_Epsilon 2个保守结构域,属于类硫氧还蛋白和谷胱甘肽S–转移酶2个超家族。系统进化树分析表明,舞毒蛾Ld GST蛋白属于GST Epsilon家族。蛋白结构显示,Ld GSTe1蛋白包含一个N端和一个C端结构,α–螺旋与β–折叠为主要结构元件。实时荧光定量PCR结果表明,在4.0、10.0 mg/L鱼藤酮药剂处理下,舞毒蛾3龄幼虫Ld GSTe1基因表达先下降后上升,推测该基因参与舞毒蛾解毒应答。     

家蚕谷胱甘肽-S-转移酶E4基因的组织表达和序列特征及重组蛋白的酶活性分析

昆虫体内的谷胱甘肽-S-转移酶(GSTs)主要行使对异生或内源有毒物质进行代谢解毒的功能。选择家蚕谷胱甘肽-S-转移酶E4的编码基因Bmgste4为靶标,研究其组织表达谱、序列结构特征及原核表达重组蛋白的酶活性。RT-PCR检测不同组织表达特征的结果显示,Bmgste4基因在家蚕5龄第3天幼虫的头部、触角、下颚须、表皮、脂肪体、丝腺以及雄性个体的马氏管和雌性个体的卵巢中均有表达,而在血细胞、中肠以及雄性个体的精巢和雌性个体的马氏管中不表达。多序列比对结果显示BmGSTE4具有完整和保守的GST二级结构特征,N端氨基酸残基保守,特别是谷胱甘肽结合位点。构建插入Bmgste4基因开放阅读框片段的重组表达质粒p28-Bmgste4,并转化E.coli Rosetta(DE3)感受态细胞进行原核表达后,利用分光光度法测定重组蛋白对通用底物1-氯-2,4-二硝基苯的酶活参数:Km=0.58 mmol/L,Vmax=24.55μmol/(mg.min)。这些结果为进一步研究家蚕体内解毒和抗药性机制以及开发鳞翅目害虫新型生物防治药剂提供了基础信息。     

Effects of UTI on oxidative stress damage in rabbit brain tissues after cardiopulmonary resuscitation

AIM: To investigate the effects of ulinastatin (UTI) on oxidative stress damage in rabbit brain tissues after cardiopulmonary resuscitation (CPR). METHODS: The model of cardiac arrest (CA) in adult male New Zealand rabbits was established and the levels of malondialdehyde (MDA) and glutathione (GSH) in plasma 24 h after restoration of spontaneous circulation (ROSC) were detected. In another experiment, 48 CA New Zealand rabbits were used. After ROSC, the animals were randomly divided into model group and UTI treatment group. The effects of UTI on MDA, GSH and the apoptosis of neurons in the cortex and hippocampus were observed. The effect of UTI on the expression of nuclear factor E2-related factor 2 (Nrf2) was also analyzed. RESULTS: The concentration of MDA in plasma increased rapidly after CA and peaked at 2 h after ROSC, whereas the content of GSH dropped to a lower level. UTI restrained the rise of MDA in the cortex and hippocampus and increased the level of GSH. UTI also accelerated the activation of Nrf2. In the cortex and hippocampus CA1~CA3 areas, the number of apoptotic neurons in model group was more than that in UTI group. CONCLUSION: UTI evidently increases the expression of Nrf2 in rabbit brain tissues, elevates the level of GSH and decreases the concentration of MDA to protect cortical and hippocampal neurons.     

Promoting oocyte aging with abnormal mitochondrial function and antioxidant post ovulation in obesity mouse

AIM: To investigate the effects of obesity on mouse oocyte in vivo aging, its in vitro fertilization and embryo development. METHODS: The oocyte-cumulus granulosa cell complexes were obtained at the time points of 14 h, 18 h and 22 h after HCG injection. The in vitro fertilization and embryo culture were conducted. Hoechst 33342 and propidium iodide double staining were performed to determine the apoptotic rate of cumulus granulosa cells. JC-1 and DCFH-DA were used to detect the levels of mitochondrial membrane potential(MMP) and reactive oxygen species (ROS), respectively. The levels of ATP and glutathione (GSH) in oocytes were also detected. RESULTS: The GSH level in obese mouse oocytes just after ovulation was higher than that in control group (P<0.05). No difference of blastocyst formation rate, ATP, ROS and cumulus granulosa cell apoptotic rate between obesity group and control group was observed (P>0.05). However, 18 h after HCG injection, ATP content and blastocyst cell numbers in obese mouse oocytes were less than those in control group (P<0.05), while ROS level in oocytes and the apoptotic rate of cumulus granulosa cells were higher than those in control group (P<0.05). Compared with control group, MMP and blastocyst formation rate were significantly decreased in obesity group 22 h after HCG injection (P<0.05). CONCLUSION: Obesity promotes oocyte aging after ovulation, and oxygen free radicals may play an important role in obesity-induced decrease of maternal infertility.