镉诱发肝细胞毒性和胞内Ca2+变化及硒的保护作用研究

本文通过研究镉诱发鼠肝细胞毒性和胞内游离Ca2 变化及硒的干预效应,探讨镉致肝细胞损伤机制及硒的保护作用。分离培养新生鼠原代肝细胞,随机分为正常对照组、4个5、25、100和250μmol/LCdCl2组、2个10和20μmol/LNa2SeO3组和8个用10和20μmol/LNa2SeO3分别与5、25、100和250μmol/LCdCl2联合作用组。在实验后第12h检测肝细胞存活及其MDA含量和培养液中LDH活性,激光共聚焦显微镜分析肝细胞内游离Ca2 水平([Ca2 ]i)。结果显示,镉处理的肝细胞存活随镉浓度增加明显下降,硒处理组与对照组差异不明显;硒提高或明显提高镉染毒肝细胞存活。肝细胞培养上清液LDH活性随镉浓度增加而逐渐升高,且100和250μmol/LCdCl2组显著高于对照组,而硒处理组未见明显变化;给予硒的25、100和250μmol/LCdCl2处理组LDH活性下降或明显下降。不同浓度镉均诱发肝细胞MDA含量显著升高,而硒处理组未见类似表现;10和20μmol/LNa2SeO3抑制或显著地降低25、100和250μmol/LCdCl2诱发的MDA的生成。经镉处理的肝细胞[Ca2 ]i荧光强度明显高于对照组,且随镉浓度的增加而上升,而给予硒的肝细胞[Ca2 ]i荧光强度未见升高,与对照组相近;加入硒的镉染毒肝细胞[Ca2 ]i均比各对应浓度的镉处理组有较大幅度地下降,其中给予硒的25μmol/LCdCl2处理组差异显著,且接近对照组的水平。结果提示,镉诱发肝细胞毒性和损伤以及肝细胞[Ca2 ]i升高;硒可能通过干预肝损伤细胞脂质过氧化反应,改善和保护肝细胞[Ca2 ]i稳态而减轻镉诱发的细胞毒性和损伤过程。     

脱落酸与镉处理对辣椒果实中辣椒素和维生素C含量的影响

本文以湘研15号辣椒为材料,研究了脱落酸（ABA）与镉（Cd^2＋）在辣椒米实不同发育时期对辣椒素和维生素C含量的影响,并分析了处理后辣椒素、维生素C含量变化与POD活性以及叶片光合特征之间的关系。结果表明：湘研15号辣椒植株经ABA与Cd^2＋处理后果实中辣椒素含量峰值分别比对照提高了80．4％与61．4％,POD活性均显著升高,二者处理后辣椒果实中的辣椒素、POD变化趋势基本一致,处理后叶片光合速率均有下降趋势。ABA处理后果实中维生素C含量在转色期比对照降低12．8％～21．7％,叶片中叶绿素相对值显著增加;Cd^2＋处理后果实中维生素C含量在转色期比对照减少18％～25％,红熟期减少13％-26％,叶片叶绿素相对值在红熟期之前比CK显著降低。     

维生素C对镉负荷小鼠睾丸抗氧化功能的影响*

目的: 分析镉(Cd)负荷不同时间对小鼠睾丸抗氧化酶的影响及维生素C(VC)的保护作用。方法: 清洁级雄性昆明小鼠72只分为4组(n=18):对照组、Cd组(CdCl₂ 3 mg/kg)、VC组(200 mg/kg)、VC(200 mg/kg)+ Cd(CdCl₂ 3 mg/kg)组,每日染毒1次,染毒1 d和3 d及同时补充VC保护,第1日和第3日染毒24 h后,每组取半数小鼠称重,取血清和睾丸组织;检测睾丸脏器系数,血清和睾丸组织丙二醛(MDA)、超氧化物歧化酶(SOD),及睾丸组织谷胱甘肽过氧化物酶(GSH-Px)、还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)及总谷胱甘肽(T-GSH)。结果: 与对照组比较,Cd组1 d和3 d小鼠体重和睾丸脏器系数下降;染毒3 d,Cd组小鼠血清SOD显著降低、MDA显著升高(P＜0.05);Cd组1 d小鼠睾丸的SOD、GSH-Px、T-GSH及GSH/GSSG显著升高(P＜0.05),而3 d的上述指标均显著降低(P＜0.05),Cd组1 d和3 d MDA水平均显著升高(P＜0.05);VC处理后减轻的程度有所降低。与Cd组比较,VC+ Cd组血清SOD和MDA水平在染毒3 d变化有显著性差异(P＜0.05);VC+ Cd组在染毒1 d和3 d,小鼠睾丸的SOD、GSH-Px、T-GSH及GSH/GSSG水平变化有显著性差异(P＜0.05),VC+ Cd组在染毒3 d睾丸的MDA水平显著降低(P＜0.05)。与Cd组1 d比较,染毒3 d小鼠的血清SOD水平显著降低(P＜0.05),睾丸指标变化也有显著性差异(P＜0.05)。结论: VC处理可在一定程度上改善镉负荷小鼠的抗氧化功能,对睾丸氧化损伤具有保护作用。     

维生素E、微量元素锌和硒对大鼠体外原代肝细胞及储脂细胞胶原代谢的影响

对维生素E、微量元素锌和硒对大鼠肝脏细胞胶原代谢的调控进行了体外研究。结果表明:肝细胞和储脂细胞均参与肝脏的胶原代谢;环境中维生素E浓度的提高可抑制肝细胞和储脂细胞的核酸合成,对细胞蛋白合成的促进作用不明显,但对细胞胶原蛋白的合成却有选择性抑制作用;环境中微量元素锌和硒浓度的适当提高对肝细胞和储脂细胞的核酸增殖和蛋白质合成均有明显促进作用,但对细胞胶原蛋白合成具有相对抑制作用,若锌和硒的浓度过高则对细胞有损害。总之,营养环境的不同将大大影响肝脏胶原的代谢,因此对肝病病人给予合理的营养膳食可能有助于防止肝硬化的恶化。     

饲料硒和维生素E对大鼠机体抗氧化能力的影响

 用克山病病区粮配成基础低硒饲料,补充硒和/或维生素E组成四种不同水平的饲料,饲喂雄性断乳大鼠,观察其对机体抗氧化能力的影响。评价指标是用抗坏血酸诱发的红细胞溶血率、被O~-_2(超氧阴离子)氧化的血红蛋白量和组织中的TBA值。动物饲养13周后,自尾静脉取血,测定溶血率和血红蛋白被氧化的百分率,和全血SeGSHPx(含硒谷胱甘肽过氧化物酶)活力。15周后将动物断头杀死,立即取出心脏和肝脏测定SeGSHPx活力和TBA值。结果表明在克山病病区粮的饲料中补充硒或维生素E,或者二者同时补充均明显提高组织中的SeGSHPx活力和降低组织中的TBA值。不论在硒缺乏时或硒充足时,饲料中补充维生素E显著降低抗坏血酸诱发的红细胞溶血率,对O~-_2氧化血红蛋白无保护作用。在维生素E缺乏时,仅补充硒对溶血无作用。不论饲料中维生素E缺乏或者充足,补充硒对O~-_1氧化血红蛋白均有显著保护作用。     

饲料中不同维生素C含量对长吻鮠的影响

用维生素C含量为38、364 和 630 mg /kg 的三种饲料饲喂初体重为23.19±0.58g的长吻鮠8个月,实验结果表明,38mg/kg Vc饲料组饲喂的长吻鮠表现出了典型的Vc缺乏症状：体色发黑,脊椎侧弯,鳍边由开始的萎缩直到腐烂;贫血;特定生长率显著降低（p＜0.05）。并且38mg/kg Vc饲料组长吻鮠的生理指标也受到影响：血清溶菌酶活性显著下降（p＜0.05）;肝脏的SOD活性及MDA含量均显著升高（p＜0.05）;血清皮质醇没有显著差异（p>0.05）;但各处理组长吻鮠肝脏中均未检测到诱导型HSP70的存在。因此,相对于皮质醇和HSP70,血清溶菌酶、肝脏SOD活性、肝脏MDA含量以及血液理化指标（血红细胞数及血红蛋白）更能反映出Vc缺乏下长吻鮠的生理状态。     

硒镉处理对萝卜硒镉吸收的影响及其交互作用

采用盆栽试验的方法,研究不同含量的硒、镉单一和复合作用对萝卜生长及其对硒、镉吸收的影响。结果表明:适量的硒(2.5~5.0 mg·kg-1)可以促进萝卜的生长,表现为萝卜发芽率提高43%、叶片SPAD值提高22%、生物量增加50%;镉单一污染时,镉显著抑制萝卜的生长(P0.05);土壤中硒的浓度低于1.5 mg·kg-1时对镉含量5.0 mg·kg-1具有拮抗作用,同时可以促进萝卜的生长;萝卜中硒的浓度随着土壤中硒的添加量的增加而增加,对硒的富集效果明显,且地下部分对硒的吸收能力大于地上部分;土壤中硒浓度5.0 mg·kg-1时,5.0 mg·kg-1镉的存在不影响萝卜对硒的吸收;当土壤中硒的浓度为5.0~10.0 mg·kg-1时,镉的存在虽然影响萝卜对硒的吸收,但萝卜中硒的含量仍处于稳定积累状态;而当土壤中硒浓度≥10.0 mg·kg-1时,镉的存在会抑制萝卜对硒的吸收,且这种作用表现为地上部分强于地下部分;硒能显著降低萝卜对镉的吸收,且这种作用表现为地下部分强于地上部分。因此,在轻度镉污染的土壤中种植富硒萝卜,既能生产营养安全的富硒食品,又可实现污染农田的再利用。     

硒对高镉引起的心肌细胞生物电活动变化的抑制作用

已经证明 ,缺硒 (Ｓｅ)是导致克山病的基本原因 ,但另外的研究进一步发现 ,单纯缺Ｓｅ并不能完全解释克山病的许多流行病学特点。研究发现 :克山病病区水、土、粮食及人群的血清、头发中除低Ｓｅ外 ,还常伴有镉 (Ｃｄ)含量较高 ,因而推测 :高Ｃｄ是导致克山病心肌损害的重要协同因素 ,为进一步证实上述观点 ,本研究在高Ｃｄ条件下培养大鼠心肌细胞 ,并以细胞内微电极技术对其生物电活动进行定量测定 ,为进一步阐明克山病心肌细胞损害的机理提供实验依据。1　材料与方法(1)动物及药品　乳鼠 (山东省实验动物中心提供 ) ,亚硒酸钠 (北京中联化…     

镉胁迫下硒对罗汉果组培苗光合特性的影响

实验以罗汉果组培苗为材料,室内栽培在内装市售营养土的塑料盆中,以0、10、50、100、200mg·kg-1浓度镉离子和1mg·kg-1浓度硒处理,培养20d后分析罗汉果幼苗的相关光合生理指标。结果表明:低浓度Cd2+对叶片叶绿素含量、光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)影响不大或稍有上升,但高浓度镉离子处理植株叶片的叶绿素含量、光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)明显下降;随Cd2+处理浓度的增加,叶片胞间CO2浓度(Ci)呈现上升趋势;加硒则延缓叶绿素下降,促进光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)上升,降低叶片胞间CO2浓度(Ci)。表明高浓度镉离子的毒害导致罗汉果组培苗叶片光合性能受到伤害,从而影响罗汉果幼苗生长。镉硒混合处理反映出硒对镉的毒害有缓解作用。     

Effects of selenium on endothelial dysfunction and metabolic profile in low dose streptozotocin induced diabetic rats fed a high fat diet

Endothelial dysfunction develops as a result of oxidative stress and is responsible for diabetic vascular complications. We investigated the effects of selenium on endothelial dysfunction and oxidative stress in type 2 diabetic rats. Male Wistar rats were divided into five groups: controls, untreated diabetics, and diabetics treated with 180, 300, 500 mcg/kg selenium each day. Diabetes was induced by a single intraperitoneal injection of low dose streptozotocin to rats fed a high fat diet. Endothelium-dependent and -independent relaxations were measured in the thoracic aorta. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and endothelial nitric oxide synthase (eNOS) mRNA expressions were analyzed using real-time polymerase chain reaction (RT-PCR). Fasting blood glucose, lipid profile, lipid oxidation, insulin and nitric oxide were measured in blood samples. Malondialdehyde, superoxide dismutase, catalase and glutathione peroxidase levels were measured in liver samples. RT-PCR showed that selenium reversed increased NADPH oxidase expression and decreased eNOS expression to control levels. Selenium also improved the impairment of endothelium-dependent vasorelaxation in the diabetic aorta. Selenium treatment significantly decreased blood glucose, cholesterol and triglyceride levels, and enhanced the antioxidant status in diabetic rats. Our findings suggest that selenium restores a normal metabolic profile and ameliorates vascular responses and endothelial dysfunction in diabetes by regulating antioxidant enzyme and nitric oxide release.     

Influence of vitamin C on cadmium absorption and distribution in rats

The purpose of the present study was to evaluate the effect of a dietary vitamin C supplement on cadmium absorption and distribution in an animal model. An aqueous solution of cadmium chloride (labelled with cadmium-109) was given by gavage to male Wistar rats for 28 days at a daily dose corresponding to 10 mg Cd/kg diet (1.0-1.2 mg Cd/kg b.w.). The animals assigned to groups 1 and 2 (45 animals per group) received a standard laboratory diet LSM, and tap water or tap water supplemented with ascorbic acid (1.5 mg/l), respectively. The radioactivity of the samples was measured using a liquid scintillation counter (tissue samples) and a gas-flow automatic counter (ashed carcasses). The fractional uptake of cadmium-109 in the carcass and organs was evaluated within 32 days after treatment by dividing the cadmium-109 activity in the whole sample by the total activity of cadmium-109 administered for 28 days. Results were compared using AUC (areas under the concentration time curve) values. The vitamin C supplement decreased the carcass cadmium burden and the cadmium content in the liver, kidneys, testicles and muscles; the highest decreases were found in the testicles, the lowest ones in the muscles. In addition, the rats supplemented with vitamin C revealed an improved body weight gain during the experimental period.     

Effects of vitamin B12 on cadmium toxicity in rats

The survival rate was high among male Sprague rats treated with 0.15 mg/kg vitamin B12 (cyanocobalamine) after injection of 5 mg/kg CdCl2.H2O (LD50). The cadmium content of the liver and, for some protocols, in the kidney was significantly reduced in survivors. According to UV-V and multinuclear (1H, 13C, 31P, and 113Cd) magnetic resonance spectroscopy no direct interaction seems to take place between cyanocobalamine and CdCl2 in aqueous solution at pH 4.5. An indirect mechanism is put forward to explain the antidotal activity.     

Alterations in plasma total and high density lipoprotein cholesterol levels in hyperlipidemic rats fed diets with varied content of selenium and vitamin E

The effect of dietary selenium and vitamin E on plasma total (TC) and high density lipoprotein cholesterol (HDLC) was evaluated in 54 Sprague Dawley rats fed cholesterol/cholic acid enriched diets. Diets 1, 2, and 3 had no added selenium (low Se) and 0 (low), 60 (adequate), and 600 (high) mg/kg dL alpha tocopheryl acetate added respectively. Sodium selenite at 0.2 mg/kg (adequate Se) was added to diets 4, 5, and 6 and at 4.0 mg/kg (toxic Se) to diet 7, 8, and 9 with the same pattern of vitamin E added to the diet as described above. TC and HDLC were measured using the Kodak Ectachem system. Rats in the low and adequate Se groups fed high vitamin E had lower TC values than rats fed lower vitamin E levels but differences were not significant. In the toxic Se groups, rats fed high vitamin E had significantly (p<0.05) higher plasma TC values than did lower Vitamin E groups. Rats on the high vitamin E diets with low or adequate Se had significantly (p<0.05) higher mean plasma HDLC values when compared to rats fed low or adequate vitamin E diets. HDLC values for animals on Se toxic diets were significantly (p<0.05) lower in rats fed a low vitamin E diet. In rats fed Se deficient and adequate diets, a high vitamin E intake resulted in a decrease in TC and an increase in HDLC. In Se toxic rats, TC was elevated by a high dietary intake of vitamin E as was HDLC with both values being significantly higher than values found in the vitamin E deficient rats. Vitamin E deficiency resulted in a plasma lipid pattern that has been associated with greater cardiovascular disease risk.     

Effects of dietary vitamin E and selenium on antioxidative defense mechanisms in the liver of rats treated with high doses of glucocorticoid

The aim of this work was to determine the effects of dietary intake vitamin E and selenium (Se) on lipid peroxidation as thiobarbituric acid reactive substances (TBARS) and on the antioxidative defense mechanisms in the liver of rats treated with high doses of prednisolone. Two hundred fifty adult male Wistar rats were randomly divided into five groups. The rats were fed a normal diet, but groups 3, 4, and 5 received a daily supplement in their drinking water of 20 mg vitamin E, 0.3 mg Se, and a combination of vitamin E and Se, respectively, for 30 d. For 3 d subsequently, the control group (group 1) was treated with a placebo, and the remaining four groups were injected intramuscularly with 100 mg/kg body weight (BW) prednisolone. After the last administration of prednisolone, 10 rats from each group were killed at 4, 8, 12, 24, and 48 h and the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and catalase (CAT) enzymes and the levels of glutathione (GSH) and TBARS in their livers were measured. GSH-Px, SOD, and CAT enzyme activities and GSH levels in prednisolone-treatment group (group 2) began to decrease gradually at 4 h, falling respectively to 38%, 55%, and 40% of the control levels by 24 h, and recovering to the control levels at 48 h. In contrast, prednisolone administration caused an increase in the hepatic TBARS, reaching up to four times the levels of the control at 24 h. However, supplementation with vitamin E and Se had a preventive effect on the elevation of the hepatic TBARS and improved the diminished activities of the antioxidative enzymes and the levels of GSH. Therefore, the present study demonstrates the effectiveness of vitamin E and Se in reducing hepatic damage in glucocorticoid-treated rats and suggests that reductions in increased TBARS as a result of prednisolone may be an important factor in the action of vitamin E and Se.     

Effects of inorganic selenium compounds on oxidative DNA damage

Exposure of Escherichia coli or mammalian cells to H2O2 results in cell death due to iron-mediated DNA damage. Since selenium compounds have been examined for their ability to act as antioxidants to neutralize radical species, and inorganic selenium compounds are used to supplement protein mixes, infant formula, and animal feed, determining the effect of these compounds on DNA damage under conditions of oxidative stress is crucial. In the presence of Fe(II) and H2O2, the effects of Na2SeO4, Na2SeO3, SeO2 (0.5-5000 microM), and Na2Se (0.5-200 microM) on DNA damage were quantified using gel electrophoresis. Both Na2SeO4 and Na2Se have no effect on DNA damage, whereas SeO2 inhibits DNA damage and Na2SeO3 shows antioxidant or pro-oxidant activity depending on H2O2 concentration. Similar electrophoresis experiments with [Fe(EDTA)](2-) (400 microM) and Na2SeO3 or SeO2 show that metal coordination by the selenium compound is required for antioxidant activity. In light of these results, Na2SeO4 may be safer than Na2SeO3 for nutritional supplements.     

Synergistic effect of vitamin C on DNA damage induced by cadmium.

Salts of divalent cadmium are well-known human mutagens and carcinogens. In the present work, the ability of vitamin C to modulate genotoxic effects of cadmium chloride on human lymphocytes was assessed using single cell gel electrophoresis (comet assay). Vitamin C at 20 and 100 micromol/l and cadmium at 5, 30 and 150 micromol/l significantly increased the tail moment of lymphocytes. Vitamin C also increased the tail moment of cells exposed to cadmium. This effect was concentration-dependent: the higher the vitamin C concentration the greater the tail moment. The combined effects of cadmium and vitamin C were more pronounced at all concentrations tested than the sum of the effects of the compounds applied separately (p < 0.05), so cadmium and vitamin C can be considered to have synergistic effects. The results obtained can be partly explained by the participation of cadmium in the Fenton reaction and reduction of its oxidized form by vitamin C.     

Effects of additional vitamin E and selenium supply on antioxidative defence mechanisms in the kidney of rats treated with high doses of glucocorticoid

The aim of this work was to determine the effects of dietary vitamin E and selenium (Se) on lipid peroxidation as thiobarbituric acid reactive substances (TBARS) and on the antioxidative defence mechanisms in the kidney of rats treated with high-doses of prednisolone. Two hundred and fifty adult male Wistar rats were randomly divided into five groups. The rats were fed a normal diet, but groups 3, 4, and 5 received a daily supplement in their drinking water of 20 mg vitamin E, 0.3 mg Se, and a combination of vitamin E and Se, respectively, for 30 days. For 3 days subsequently, the control group (group 1) was treated with a placebo, and the remaining four groups were injected intramuscularly with 100 mg kg(-1) body weight (bw) prednisolone. After the last administration of prednisolone, 10 rats from each group were killed at 4, 8, 12, 24, and 48 h and the activities of glutathione peroxidase (GSH-Px) and catalase (CAT) enzymes, and the levels of glutathione (GSH) and TBARS in their kidneys were measured. GSH-Px and CAT enzyme activities and GSH levels in the prednisolone treatment group (group 2) began to decrease gradually at 4 h, falling respectively to 48 and 65% of the control levels by 24 h, and recovering to the control levels at 48 h. In contrast, prednisolone administration caused an increase in TBARS in the kidneys, reaching up to twice the levels of the control group at 24 h. However, supplementation with vitamin E and Se had a preventive effect on the elevation of kidney TBARS and improved the diminished activities of the antioxidative enzymes and the levels of GSH. Therefore, the present study demonstrates the effectiveness of vitamin E and Se in reducing kidney damage in glucocorticoid-treated rats and suggests that reductions in increased TBARS due to prednisolone may be an important factor in the action of vitamin E and Se.