顺铂诱导非洲绿猴肾细胞凋亡模型的建立

目的:建立常见凋亡诱导剂顺铂(Cisplatin)诱导非洲绿猴肾细胞(Vero)凋亡的模型,为进一步研究抗凋亡基因在细胞凋亡中的分子机理打下基础。方法:分别以不同浓度的顺铂处理Vero细胞48h,用噻唑蓝(MTT)比色法、Gimesa染色、流式细胞术检测,观察处理后细胞的生长活力和凋亡情况。结果:以未处理的细胞作为对照组,1、2、3、4、5μg/mL的顺铂处理的Vero细胞生存率分别为(79.02±6.10)%、(68.84±4.42)%、(56.66±4.07)%、(46.83±3.76)%、(29.04±5.93)%(P<0.01);经顺铂诱导后细胞形态学发生明显改变,出现膜小泡和凋亡小体形成等凋亡细胞特征;流式细胞仪检测,0、1、2、3、4、5μg/mL的顺铂处理的Vero细胞后凋亡率分别为1.66%±0.19%、16.65%±1.26%、24.82%±1.03%、36.22%±1.04%、48.49%±1.24%、43.34%±1.17%(P<0.01)。结论:本实验成功建立顺铂诱导非洲绿猴肾细胞凋亡模型,将有助于进一步探讨目的基因在Vero细胞凋亡作用的的分子机制。     

顺铂诱导非洲绿猴肾细胞凋亡模型的建立

目的：建立常见凋亡诱导剂顺铂（Cisplatin）诱导非洲绿猴肾细胞（Vero）凋亡的模型,为进一步研究抗凋亡基因在细胞凋亡中的分子机理打下基础。方法：分别以不同浓度的顺铂处理Veto细胞48h,用噻唑蓝（MTT）比色法、Gimesa染色、流式细胞术检测,观察处理后细胞的生长活力和凋亡情况。结果：以未处理的细胞作为对照组,1、2、3、4,5μg／mL的顺铂处理的Vero细胞生存率分别为（79．02±6．10）％、（68．84±4．42）％、（56．66±4．07）％、（46．83±3．76）％、（29．04±5．93）％（P〈0．01）;经顺铂诱导后细胞形态学发生明显改变,出现膜小泡和凋亡小体形成等凋亡细胞特征;流式细胞仪检测,0、1、2、3、4、5μg／mL的顺铂处理的Vero细胞后凋亡率分别为1．66％±0．19％、16．65％±1．26％、24．82％±1．03％、36．22％±1．04％、48．49％±1．24％、43．34％±1．17％（P〈0．01）。结论：本实验成功建立顺铂诱导非洲绿猴肾细胞凋亡模型,将有助于进一步探讨目的基因在Vero细胞凋亡作用的的分子机制。     

顺铂致肾损伤时血管紧张素Ⅱ的变化

目的：探讨顺铂致大鼠肾损伤时血管紧张素Ⅱ含量的变化。方法：取24只大鼠随机分为2组（N=12）：正常组、模型组。采用顺铂尾静脉注射的方法复制顺铂肾损伤模型。6周后,放射免疫法检测肾脏AngⅡ水平,Nasson染色测定肾脏胶原含量,计算肾脏指数。结果：与正常组相比,模型组大鼠实验末体重明显下降、肾脏指数增高,肾脏胶原含量升高、肾组织AngⅡ含量增加（P〈0．01）。结论：AngⅡ可能在顺铂肾损伤的发生、发展中起一定作用。     

抗癌药物顺铂对DNA结构的影响

本文采用脉冲激光荧光技术及常规的荧光分光光度计详细地研究了顺铂对DNA变性温度的影响,DNA氢键受损的碱基对数随温度变化,DNA的G—C碱基含量对顺铂作用效果以及顺铂浓度的影响,初步探讨了抗癌药物顺铂对DNA的结构影响.     

卡维地洛对顺铂所致大鼠肾毒性预防作用的形态学研究

目的研究顺铂诱导大鼠肾损伤的组织病理学和超微结构变化以及卡维地洛对大鼠肾损伤的预防作用。方法雄性Wistar大鼠随机分为4组,给药后3～6d处死,取肾组织,常规石蜡切片,HE染色、PAS反应,超薄切片电镜观察。结果卡维地洛组较顺铂肾脏损伤模型组的组织病理学及超微结构变化明显减轻。结论卡维地洛预防性灌胃给与能明显减轻大鼠顺铂所致的肾损伤,其机制可能与其抗氧化和清除自由基活性有关。     

内毒素在大剂量顺铂所致大鼠急性肾损伤过程中的变化及意义

目的利用大剂量顺铂（cisplatin,DDP）所致大鼠急性肾功能衰竭的动物模型,观察外周血内毒素（endotoxin）在大鼠急性肾损伤中的变化及其意义。方法SD大鼠36只,雌雄各半,依体重随机分为DDP用药6h、48h、对照组和生理盐水（NS）用药6h、48h、对照组,每组6只。10mg／kgDDP单次腹腔内注射,等量Ns对照。观察并记录用药后对照组大鼠的毒副反应;用药6、48h各组大鼠无菌条件下心脏穿刺取血、肝素抗凝,检测外周血内毒素含量,同时内眦静脉取血,测定血清尿素氮、肌酐浓度,并进行统计学分析。结果DDP用药后6h,大鼠体重开始明显降低,用药48h后,大鼠腹泻逐渐加重,用药3d后大鼠死亡。DDP用药后6h大鼠血尿素氮、肌酐的含量与对照组比较差异无显著性（P〉0．05）;DDP用药后48h血尿素氮升至（18．71±9．9）mmol／L,明显高于对照组（7．48±0．6）mmol／L（P〈0．05）,同时血肌酐含量亦升至（49．6±14．1）μmol／L,与对照组（27．17±1．7）μmol／L比较差异具有显著性（P〈0．05）。DDP用药后6h所有大鼠外周血内毒素含量都低于0．0218Eu／rrd最低检出限,明显低于NS对照组大鼠（0．3141±0．1477）Eu／ml（P〈0．01）;DDP用药后48h大鼠外周血内毒素的含量增高均超过0．70Eu／ml最高检出限,明显高于NS对照组大鼠（0．1661±0．1198）Eu／ml（P〈0．01）。结论外周血内毒素含量的变化与大剂量顺铂所致大鼠急性肾损伤早期的发病机制无关,但与大鼠肾功能衰竭有关的发生相关。     

加味黄连阿胶汤对顺铂致肾损伤大鼠TGF-β_1表达的影响

目的：观察加味黄连阿胶汤对顺铂致肾损伤大鼠TGF-β1表达的影响。方法：采用尾静脉注射顺铂的方法,制备顺铂肾毒性大鼠模型,然后给以相应的药物治疗,连续8周。剖取肾脏行HE、免疫组织化学染色,观察肾组织TGF-β1的表达情况,测定其平均吸光度。结果：模型组大鼠24h尿蛋白定量、NAG含量及TGF-β1的平均吸光度均明显高于正常对照组（P〈0.01）,而各治疗组均明显低于模型组（P〈0.05或P〈0.01）。结论：加味黄连阿胶汤可明显下调肾小管上皮细胞内TGF-β1的表达,减轻顺铂引起的肾小管和肾小管间质损伤。     

生物系统中脂质过氧化检测方法的评述

生物体系中脂质过氧化的检测方法是脂质过氧化反应机理研究能否取得成功的关键因素之一.检测生物体系中脂质过氧化的方法有多种,但各种检测方法在不同的实验中都显示出一定的优缺点,近年来应用较多并且很有前途的实验方法是高压液相色谱法以及化学发光法等.     

顺铂肾损伤机制的研究及进展

顺铂(cDDP)是临床化疗最常用的药物之一.它是一种广谱抗肿瘤药,对实体肿瘤的疗效尤为明显,其疗效与剂量成正比.     

在96孔板中进行抗脂质过氧化的微量测定

以Fe²⁺/半胱氨酸诱导大鼠肝微粒体为基本模型,根据硫代巴比妥酸(TBA)反应原理,优化不同反应条件,建立了一种在96孔板上进行抗脂质过氧化测定的一步反应方法,该方法的灵敏度不低于传统的试管法,而且还具有微量、快速、简便等优点,特别适用于大规模筛选和研究抗氧化剂.此外,也可用于其它系统诱导的抗脂质过氧化的测定.     

Cadmium induced lipid peroxidation in rat testes and protection by selenium

The main goal of this study was to investigate the role of cadmium in the promotion of lipid peroxidation in the homogenates of rat testes and the effect of selenium on lipid peroxidation in testes of rats after cadmium injection. Treatment of rats with cadmium resulted in a time- and dose-related accumulation of the metal ions in testes. The concentrations of cadmium, copper, zinc, selenium and iron in the tissues were determined by an atomic absorption spectrophotometer and lipid peroxidation in testes was measured by a spectrophotometer. Cadmium produced enhanced lipid peroxidation in testes. These cadmium-induced changes were accompanied by a significant increase of iron and copper, and a decrease of zinc in testes. Concurrent treatment with selenium and cadmium reduced the cadmium-induced alterations in lipid peroxidation and essential metal levels. Data suggest that lipid peroxidation was associated with cadmium toxicity in testes and that the addition of selenium was found to be effective in attenuation of this effect.     

Effects of cadmium-metallothionein on renal organic ion transport and lipid peroxidation in rats

The effects of cadmium-metallothionein (Cd-MT) on organic ion uptake in renal cortical slices and lipid peroxidation in the kidney were studied in rats. For in vitro studies, slices were prepared from kidneys of control animals and incubated in buffer containing either cadmium chloride (CdCl₂) or Cd-MT in equimolar Cd concentrations ranging from 5 × 10^?6 to 2 × 10^?4 M. Uptake into the slices of the organic anion p-aminohippuric acid (PAH) was found to be inhibited by both forms of Cd in a dose-dependent manner. Although this inhibition was slightly greater in the presence of Cd-MT, accumulation of Cd into the slices was approximately 12 times greater with CdCl₂ than Cd-MT. Tetraethylammonium (TEA) uptake was less sensitive to the inhibitory effects of both CdCl₂ and Cd-MT, although a dose-dependent inhibition did occur with higher Cd concentrations. To study the in vivo effects of Cd-MT on transport function and lipid peroxidation in the kidney, rats were injected with Cd-MT (0.3 mg Cd per kilogram body weight [bw]) and sacrificed at specific time intervals. Similar to the in vitro studies, PAH uptake into the renal cortical slices was markedly inhibited within 12 hours after Cd-MT injection whereas inhibition of TEA uptake was less and not observed until 48 hours after injection. Only a small increase (1.4-fold) in lipid peroxidation, as measured by generation of malondialdehyde (MDA), in the kidney was detected at four hours postinjection, and no further increase was observed at later time periods. The results suggest that Cd-MT affects the transport of organic anions and cations during its renal uptake but that lipid peroxidation may play only a minor role in Cd-MT-induced renal toxicity.     

Lipid peroxidation in liver of rats administrated with methyl mercuric chloride

Parenteral administration of methyl mercuric chloride (MMC, CH₃HgCl) to rats enhanced lipid peroxidation in liver of rats, as measured by the thiobarbituric acid reaction for malondialdehyde (MDA) in fresh tissue homogenates. After sc injection of CH₃HgCl (5 mg/kg body wt), MDA concentration in liver became significantly increased at 24 h and further increased at 48 h. Dose-response studies were carried out with male albino rats of the Fisher-344 strain (body wt 170–280 g) injected with 3 or 5 mg Hg/kg as CH₃HgCl and sacrificed after 24 h. In time-response studies, animals were administered 5 mg Hg/kg as CH₃HgCl and sacrificed after 24 and 48 h. Studies in the authors’ laboratory have shown that (1) mercury is accumulated in liver; (2) concentration of MDA is increased in liver of CH₃HgCl-treated rats; (3) severity of hepatotoxicity is generally proportional to the elevation of MDA concentration, based upon the dose-effect relationships observed after administration of CH₃HgCl to rats. The results of this study implicate that the lipid peroxidation is one of the molecular mechanisms for cell injury in acute CH₃HgCl poisoning.     

Formation of active oxygen species and lipid peroxidation induced by hypochlorite.

Hypochlorite or its acid, hypochlorous acid, may exert both beneficial and toxic effects in vivo. In order to understand the role and action of hypochlorite, the formation of active oxygen species and its kinetics were studied in the reactions of hypochlorite with peroxides and amino acids. It was found that tert-butyl hydroperoxide and methyl linoleate hydroperoxide reacted with hypochlorite to give peroxyl and/or alkoxyl radicals with little formation of singlet oxygen in contrast to hydrogen peroxide, which gave singlet oxygen exclusively. Amino acids and ascorbate reacted with hypochlorite much faster than peroxides. Free radical-mediated lipid peroxidation of micelles and membranes in aqueous suspensions was induced by hypochlorite, the chain initiation being the decomposition of hydroperoxides by hypochlorite. It was suppressed efficiently by ebselen which reduced hydroperoxides and by alpha-tocopherol, which broke chain propagation, but less effectively by hydrophilic antioxidants present in the aqueous phase. Cysteine suppressed the oxidation, but it was poorer antioxidant than alpha-tocopherol. Ascorbate also exerted moderate antioxidant capacity, but it acted as a synergist with alpha-tocopherol. Taken together, it was suggested that the primary target of hypochlorite must be sulfhydryl and amino groups in proteins and that the lipid peroxidation may proceed as the secondary reaction, which is induced by radicals generated from sulfenyl chlorides and chloramines.     

Protective effect of lipoic acid on adriamycin induced lipid peroxidation in rat kidney

Adriamycin, which is widely used in the treatment of various neoplastic conditions, exerts toxic effects in many organs. The present study was designed to investigate the effect of lipoic acid upon adriamycin induced peroxidative damages in rat kidney. The increase in peroxidated lipids on adriamycin administration was accompanied by alterations in the antioxidant defense systems. The extent of nephrotoxicity induced by adriamycin was evident from the decreased activities of the enzymes -glutamyl transferase and -glucuronidase in the rat renal tissues. The study was carried out with adult male albino rats of Wistar strain, which comprised of one control and three experimental groups. Group I rats served as controls. GroupII rats received adriamycin (1 mg kg^–1 body wt day^–1) intravenously through the tail vein. Group III rats were given lipoic acid (35 mg kg^–1 body wt day^–1) intraperitoneally. Group IV rats were given lipoic acid 24 h before the administration of adriamycin. Rats subjected to adriamycin administration showed a decline in the thiol capacity of the cell accompanied by high malondialdehyde levels along with lowered activities of catalase, superoxide dismutase, glutathione peroxidase and glutathione metabolizing enzymes (glutathione reductase, glucose-6-phosphate dehydrogenase, glutathione-S-transferase). Lipoic acid pretreatment also restored the activities of -glutamyl transferase and -glucuronidase nearly to control levels thereby suggesting nephroprotection. The study has highlighted the beneficial effects of lipoic acid pretreatment in reversing the damages caused by adriamycin and thereby bringing about an improvement in the oxidative stress parameters.     

Rosmarinic acid prevents lipid peroxidation and increase in acetylcholinesterase activity in brain of streptozotocin‐induced diabetic rats

We investigated the efficacy of rosmarinic acid (RA) in preventing lipid peroxidation and increased activity of acetylcholinesterase (AChE) in the brain of streptozotocin‐induced diabetic rats. The animals were divided into six groups (n = 8): control, ethanol, RA 10 mg/kg, diabetic, diabetic/ethanol and diabetic/RA 10 mg/kg. After 21 days of treatment with RA, the cerebral structures (striatum, cortex and hippocampus) were removed for experimental assays. The results demonstrated that the treatment with RA (10 mg/kg) significantly reduced the level of lipid peroxidation in hippocampus (28%), cortex (38%) and striatum (47%) of diabetic rats when compared with the control. In addition, it was found that hyperglycaemia caused significant increased in the activity of AChE in hippocampus (58%), cortex (46%) and striatum (30%) in comparison with the control. On the other hand, the treatment with RA reversed this effect to the level of control after 3 weeks. In conclusion, the present findings showed that treatment with RA prevents the lipid peroxidation and consequently the increase in AChE activity in diabetic rats, demonstrating that this compound can modulate cholinergic neurotransmission and prevent damage oxidative in brain in the diabetic state. Thus, we can suggest that RA could be a promising compound in the complementary therapy in diabetes. Copyright © 2013 John Wiley & Sons, Ltd.     

Protective role of ascorbic acid against lipid peroxidation and myocardial injury

Ascorbic acid (AH₂) is a potential scavenger of superoxide radical and singlet oxygen. In the guinea pig, marginal AH₂ deficiency results in intracellular oxidative damage in the cardiac tissue as evidenced by lipid peroxidation, formation of fluorescent pigment and loss of structural integrity of the microsomal membranes. The oxidative damage does not occur due to lack of enzymatic scavengers of reactive oxygen species such as superoxide dismutase, catalase and glutathione peroxidase. Also, glutathione transferase activity is not decreased in AH₂ deficiency. Lipid peroxidation, fluorescent pigment formation and protein modification disappear after AH₂ therapy. These results, if extra-polated to human beings, would indicate that chronic subclinical AH₂ deficiency may result in progressive oxidative damage which in the long run may lead to permanent degenerative diseases in the heart.     

Age-dependent changes in rat liver lipid peroxidation and glutathione content induced by acute ethanol ingestion

The study of the influence of the age of animals (13 to 53 weeks) on total liver thiobarbituric acid reactive substances (TBAR) content showed an increase which is maximal in rats of 39 weeks of age compared to young animals (13 weeks), followed by a dimunition in the 53 weeks old group. In this situation, the content of hepatic GSH and total GSH equivalents as well as the GSH/GSSG ratio were decreased with ageing, while GSSG levels were enhanced in the oldest group studied. Acute ethanol intoxication resulted in a marked increase in liver TBAR content in young animals, together with a decline in GSH, total GSH equivalents and GSH/GSSG ratio, and an enhancement in GSSG. These changes elicited by ethanol intake were reduced with ageing. It is concluded that ethanol-induced oxidative stress in the liver is diminished during ageing, despite the progressive decrease in the glutathione content of the tissue observed in control animals.     

Effect of oral methionine on blood lipid peroxidation and antioxidants in alloxan-induced diabetic rats

Supplementation of thiol compounds has been suggested to protect against the toxic effects of reduced oxygen species by contributing to the thiol pool of the cell. The present study was designed to determine whether supplementation of methionine in the diet of diabetic animals protected against the oxidative stress in diabetic pathology. Oral methionine was administered at a dosage of 330 mg/100 g feed to diabetic rats. The effect was compared with the effect of insulin administration. Levels of lipid peroxides were measured in plasma, erythrocytes, and erythrocyte membrane. Anti-oxidants were measured in plasma. Diabetic condition was associated with increased lipid peroxidation and depletion in antioxidant levels. Although methionine did not affect the level of blood glucose and some of the antioxidants, it lowered the lipid peroxide content in blood. Erythrocyte lipid peroxidation activity was unaffected by methionine treatment. Administration of insulin lowered both plasma and erythrocyte lipid peroxide levels.     

Local and systemic increase in lipid peroxidation after moderate experimental traumatic brain injury

Traumatic brain injury is a common event associated with neurological dysfunction. Oxidative damage, may contribute to some of these pathologic changes. We used a specific and sensitive marker of lipid peroxidation, the isoprostane 8,12-iso-iPF(2alpha) -VI, to investigate whether local and also systemic lipid peroxidation were induced following lateral fluid percussion (FP) brain injury in the rat. Animals were anesthetized and subjected to lateral FP brain injury of moderate severity, or to sham injury as controls. Urine was collected before anesthesia (baseline), 6 and 24 h after injury. Blood was collected at baseline, 1, 6 and 24 h after injury. Animals were killed 24 h after surgery and their brains removed for biochemical analysis. No significant difference was observed at baseline (preinjury) for urine and plasma 8,12-iso-iPF(2alpha) -VI levels between injured and sham-operated animals. By contrast, plasma and urinary levels increased significantly already at 1 and further increased 24 h following brain injury, when compared to sham-operated animals. Finally, compared with sham, injured animals had a significant increase in brain 8,12-iso-iPF(2alpha) -VI levels. These results demonstrate that moderate brain injury induces widespread brain lipid peroxidation, which is accompanied by a similar increase in urine and plasma. Peripheral measurement of 8,12-iso-iPF(2alpha) -VI levels after brain injury may be a reliable marker of brain oxidative damage.