Comparison of cellular and molecular cytotoxic mechanisms of Cochlodinium polykrikoides in isolated trout and rat hepatocytes

Harmful algal blooms produced by the marine ichthyotoxic dinoflagellate Cochlodinium polykrikoides are responsible for mass mortalities of wild and farmed fish globally. This study compared the cytotoxic mechanisms of C. polykrikoides total extract on both trout and rat liver hepatocytes. Trout hepatocytes were more sensitive than rat hepatocytes against C. polykrikoides extract. The effective concentration 50 after 3 hour incubation (EC50_3hr) concentrations found for C. polykrikoides extract in trout and rat hepatocytes (i.e., 50% membrane lysis in 3 hr) were Eq. 1 cell/ml and Eq. 240 cell/ml, respectively. C. polykrikoides extract exposure in both isolated trout and rat hepatocytes resulted in membrane lysis, reactive oxygen species formation, glutathione depletion, collapse of mitochondrial membrane potential, ATP depletion, increase in adenosine diphosphate (ADP)/adenosine triphosphate (ATP) ratio, cytochrome c release into the hepatocyte cytosol, and activation of caspases cascade. Trout hepatocyte toxicity was also associated with lysosomal membrane injury. Mitochondrial permeability transition in both trout and rat hepatocytes produced cytochrome c release from the mitochondrial intramembrane space into the cytosol. Thus, the cytochrome c release triggered activation of caspase-3 and apoptosis. Finally, data demonstrated that C. polykrikoides extract may induce more apoptotic phenotype in rat than trout hepatocytes, which in the latter favored predominantly necrotic mode of cell death.     

Thallium(I) and thallium(III) induce apoptosis in isolated rat hepatocytes by alterations in mitochondrial function and generation of ROS

Environmental metal toxins, generated through diverse anthropogenic activities, constitute one of the major contaminants that have led to global dispersion of these toxic metals in the ecosystem. Thallium is one of these widely dispersed metals that produce severe adverse effects on human and biological systems. The influence of thallium(I) and thallium(III) on the early events that trigger apoptosis signaling were examined in freshly isolated rat hepatocytes. In addition, the role of oxidative stress, and mitochondria in the induction of apoptosis were also investigated. Incubation of thallium(I) and thallium(III) with isolated rat hepatocytes generated reactive oxygen species (ROS), collapse of mitochondrial membrane potential, activation of caspases cascade, and appearance of apoptosis phenotype. Mitochondrial permeability transition (MPT) pore sealing agents (cyclosporine A and carnitine) and ATP generators (L-glutamine, fructose, and xylitol) inhibited the activation of caspase-3 and apoptosis, indicating that both the cations activated apoptosis signaling via mitochondrial pathway. Pretreatment of hepatocytes with antioxidants (α-tocopherol or deferoxamine) also blocked caspase-3 activation induced by these cations, suggesting that oxidative stress may be directly involved in a mitochondrial MPT pore opening and activation of caspases cascade. These findings contribute to a better understanding of the mechanisms that mediate thallium-induced apoptosis in isolated rat hepatocytes.     

Mitochondrial toxicity and in vitro biological effects of ambient PM2.5 from an urban site in China

Abstract

The roles of PM_2.5-induced mitochondrial damage and oxidative stress on mast cell degranulation were examined in vitro. Mast cells were treated with suspensions of PM_2.5 in Dulbecco’s modified Eagle’s medium at concentrations from 25 to 200?mg/L in the absence or presence of 10?mmol/L N-acetyl-L-cysteine. Biological effects and mitochondrial function were assessed by determining cell viability, β-hexosaminidase release, interleukin-4 secretion, reactive oxygen species generation, adenosine triphosphate production, potential alteration of mitochondrial membrane, and activities of mitochondrial electron transport chain complexes I and III. Exposure of mast cells to PM_2.5 induced reduction of adenosine triphosphate production, collapse of mitochondrial membrane potential, and inhibition of the activity of complex III. Co-treatment of mast cells exposed to PM_2.5 with N-acetyl-L-cysteine attenuated cytotoxicity and the production of reactive oxygen species, and decreased the release of β-hexosaminidase and interleukin-4. Evidently, PM_2.5-induced oxidative stress plays an essential role in mitochondrial toxicity and mast cell activation.     

Antimony induces oxidative stress and cell death in normal hepatocytes

Antimony (Sb) accumulates in the liver which is one of the target organs for metal-mediated toxicity. Although toxicity of Sb was previously investigated, the precise mechanism of Sb-induced hepatotoxicity remains to be determined. The aim of this study was to examine the role of oxidative stress, and mitochondria in the induction of cell death by Sb. Our results showed that liver cell lysis induced by Sb is mediated by reactive oxygen species (ROS) formation, lipid peroxidation and decline of mitochondrial membrane potential (MMP). Antimony-induced ROS formation, lipid peroxidation and reduction of MMP were significantly diminished by antioxidants and ROS scavengers such as dimethyl sulfoxide and mannitol; mitochondrial permeability transition (MPT) pore sealing agents such as carnitine and trifluoperazine; and adenosine triphosphate (ATP) generator, L-glutamine. Antimony-induced ROS formation, lipid peroxidation and fall in MMP were potentiated by glutathione (GSH) depletion via n-bromoheptane. MPT pore sealing agents and ATP generator inhibited hepatotoxicity, indicating Sb-activated cell death via mitochondrial pathway. Pretreatment of hepatocytes with antioxidants and ROS scavengers also blocked cell death induced by Sb, whereas GSH depletion enhances Sb-induced cell death, suggesting that oxidative stress may be directly involved in the reduction of MMP. These findings contribute to a better understanding of the mechanisms that mediate Sb-induced cell death in isolated rat hepatocytes.     

Direct toxicity of amyloid beta peptide on rat brain mitochondria: preventive role of Mangifera indica and Juglans regia

Role of mitochondrial dysfunction and oxidative stress has been well documented in various cognitive-related disorders such as Alzheimer's disease (AD). Evidence indicates that Aß formation impairs mitochondrial function and that mitochondrial dysfunction is an early event in the pathogenesis of AD. The present study was, therefore, designed to investigate the direct toxicity of Aß peptide on isolated mitochondria obtained from rat brain. Various mitochondrial toxicity/integrity parameters such as succinate dehydrogenase activity, reactive oxygen species (ROS) formation, mitochondrial membrane potential collapse (MMP), mitochondrial swelling, and cytochrome c release were measured following the addition of Aß peptide on isolated mitochondria and then, mitoprotective effect of aqueous extracts of Mangifera indica and Juglans regia against mitochondrial toxicity endpoints parameters induced by Aß peptide were assessed. Our results showed that exposure to Aß peptide (30 nM) in isolated brain mitochondria induced mitochondrial ROS formation, MMP collapse, mitochondrial swelling, and cytochrome c release which is the starting point of apoptosis signaling. All these mitochondrial toxic endpoints induced by Aß peptide inhibited by aqueous extracts of Mangifera indica (100–400 µg/ml) and Juglans regia (200–400 µg/ml). To our knowledge, this is one of the first apparent studies to claim directly targeting of brain mitochondria and induction of apoptosis by Aß peptide as a new hypothesis for etiology of AD and other related neurodegenerative diseases as well as mitopreventive role of common antioxidant nutritional products including walnut and mango.     

ZnO纳米粒子通过线粒体通路抑制小鼠光感受器细胞Na~+/K~+-ATP酶表达和活性的作用研究

氧化锌(ZnO)纳米粒子已被发现具有生物毒性,氧化应激被认为是最重要的因素之一。前期实验证实,ZnO纳米粒子能显著减少锰超氧化物歧化酶(Mn SOD)蛋白的表达,降低Mn SOD活性。本文通过检测乳酸脱氢酶(LDH)释放、线粒体活性氧(ROS)水平和膜电位(Δφm)、延迟整流钾电流变化和Na~+/K~+-ATP酶的表达及活性等变化,检测ZnO纳米粒子对小鼠光感受器细胞的细胞毒作用。结果表明,ZnO纳米粒子可显著增强小鼠光感受器细胞中LDH的释放、增加线粒体内ROS水平并下调Δφm、阻断延迟整流钾电流,同时降低Na~+/K~+-ATP酶的表达及活性,从而对小鼠视网膜光感受器细胞产生细胞毒作用,提示ZnO纳米粒子可通过线粒体通路引起氧化应激,从而抑制小鼠光感受器细胞Na~+/K~+-ATP酶表达和活性,产生细胞毒性,导致细胞死亡。本文的研究结果有助于理解ZnO纳米粒子引起细胞毒性的作用机理。     

Toxicity of depleted uranium on isolated liver mitochondria: a revised mechanistic vision for justification of clinical complication of depleted uranium (DU) on liver

Depleted uranium (DU) is widely used in military anti-armor weapons. Recent evidence suggested that oxidative stress and mitochondrial dysfunction may contribute to DU-induced toxicity. However, the underlying mechanisms of DU toxicity in mitochondria are not well understood. In this study, liver mitochondria were obtained from Wistar rats treated with DU in the form of uranyl acetate (UA) (0.5, 1 or 2 mg/kg i.p.) using differential centrifugation. For in vitro experiments, control rat liver mitochondria were incubated with different concentrations of UA (50, 100 or 200 μM) for 1 hr. Mitochondrial reactive oxygen species (ROS) production, collapse of mitochondrial membrane potential, and mitochondrial swelling were examined by flow cytometry. Mitochondrial sources of ROS formation were determined using specific substrates and inhibitors. Extent of lipid peroxidation (LPO) and glutathione (GSH) oxidation, and also complex II and IV activities were detected via spectroscopy. Further, the concentration of ATP and ATP/ADP ratio was measured using luciferase enzyme and release of cytochrome c from mitochondria which was detected by ELISA kit. UA induced succinate-supported mitochondrial ROS production, elevated LPO levels, GSH oxidation, and mitochondrial complex II inhibition. UA also induced mitochondrial permeability transition and increase in cytochrome c release which subsequently disturbed oxidative phosphorylation and reduced the mitochondrial ATP concentration. Data suggest that mitochondrial oxidative stress and uncoupling of oxidative phosphorylation may play key roles in DU-induced hepatic toxicity.     

Single walled carbon nanotubes induce cytotoxicity and oxidative stress in HEK293 cells

The aim of the present study was to evaluate the potential toxicity and general mechanisms involved in single walled carbon nanotubes (SWCNTs)-induced cytotoxicity using human embryonic kidney cell line (HEK293) cells. Carbon nanotubes (coded as CNT) used in this study were synthesized by the chemical vapor deposition method. To elucidate the possible mechanisms underlying SWCNT-induced cytotoxicity, cell viability, cell membrane damage (lactate dehydrogenase activity (LDH) assay), reduced glutathione (GSH), interleukin-8 (IL-8) and lipid peroxidation products levels were quantitatively assessed following SWCNT exposure for 48 hr using HEK293 cells. Exposure of cells to SWCNT at 3–300 μg/ml produced significant reduction in cell viability in a concentration-dependent manner. The IC₅₀ value of SWCNT was found to be 87.58 μg/ml. Exposure of HEK cells to SWCNT at 10–100 μg/ml resulted in concentration-dependent cell membrane damage, increased production of IL-8, elevated levels of thiobarbituric acid reactive substances like malondialdehyde and decreased intracellular GSH levels. In summary, exposure to SWCNT resulted in a concentration-dependent cytotoxicity in cultured HEK293 cells that was associated with increased oxidative stress.     

Toxicity of 4-methylimidazole on isolated brain mitochondria: using both in vivo and in vitro methods

4-methylimidazole (4MI) is a compound widely used in various industrial and consumer applications. The most important sources of exposure include chemical caramel coloring, ammoniated molasses, dyes and pigments, rube, cleaning and agricultural chemicals. Toxicity attributed to 4MI in foods has recently become a focus of research. Recent studies showed that 4MI induced adverse changes in various target tissues. Brain is known to be a target organ for 4MI-induced toxicity but its cytotoxic mechanisms have not yet been elucidated. In this study, experiments were divided into two parts: (1) using in vivo methodology, doses of 4MI at 100, 200, or 300 mg/kg were administered orally to mice daily for 14 to obtain brain mitochondria; and (2) utilizing in vitro methodology, brain mitochondria were incubated with 4MI at 400, 800, or 1600 μM concentrations. Subsequently, the neurotoxicity of 4MI was assessed using mitochondrial dysfunction tests, including reactive oxygen species (ROS) formation, mitochondrial membrane potential (MMP) collapse, mitochondrial swelling, and cytochrome c release. Our results from both in vivo and in vitro experiments on isolated brain mitochondria showed a significant decrease in complex II activity and also marked elevation in the ROS formation, MMP collapse, mitochondrial swelling, and enhanced release of cytochrome c. Data indicated that 4MI induced neurotoxicity through the impairment of electron transfer chain especially at complex II and elevated ROS formation leading to subsequent oxidative stress events including mitochondrial membrane depolarization, mitochondrial swelling, and release of cytochrome c, which is the starting point of mitochondrial-mediated apoptosis signaling and neurodegeneration.     

Effect of Zinc Oxide nanoparticles on cellular oxidative stress and antioxidant defense mechanisms in mouse liver

Zinc oxide nanoparticles (ZnO₂), a common ingredient of cosmetics has a huge variety of applications. Previous studies reported oxidative stress mediated toxicity of ZnO₂ nanoparticles on various mammalian cell lines. Although zinc (Zn) is an essential mineral at higher concentrations this metal is toxic. The present study focused on size determination by monitoring changes in activities of antioxidant defense mechanism in response to oxidative stress induced by ZnO₂ nanoparticles using mouse liver tissue homogenates. The study also investigated effects of oxidative stress induced DNA damage by determining formation of 8-OHdG in mouse liver homogenate. A cytotoxicity assay was also carried out in L929 cells to determine cell viability. The results of the study indicated that 50μg/ml of ZnO₂ nanoparticles induced 50% cell death. Alterations in antioxidant parameters and 8-OHdG were also noted. Data showed that there was a concentration-dependent fall in cell viability, decrease antioxidant enzyme levels and increase formation of DNA adduct (8-OHdG) when mouse liver tissue homogenate were exposed to ZnO₂ nanoparticles.     

Toxicity of cigarette smoke on isolated lung,heart, and brain mitochondria: induction of oxidative stress and cytochrome c release

Cigarette smoking is one of the main risk factors for premature human death which is associated with a variety of respiratory and vascular diseases, and cancer due to exposure to hundreds of toxicants. Rat mitochondria were obtained by differential ultracentrifugation and incubated with different concentrations (1%, 10%, or 100%) of standardized cigarette smoke extract (CSE). Our results showed that CSE induced a rise in mitochondrial reactive oxygen species (ROS) formation, lipid peroxidation, and mitochondrial membrane potential (MMP) collapse before mitochondrial swelling ensued in isolated pulmonary mitochondria. Disturbance in oxidative phosphorylation was also confirmed by decrease in ATP concentration in the CSE-treated mitochondria. In addition, collapse of MMP and mitochondrial swelling produced release of cytochrome c via outer membrane rupture or mitochondrial permeability transition (MPT) pore opening. Our results suggested that CSE-induced toxicity in lung tissue is the result of disruptive effect on mitochondrial respiratory chain that leads to ROS formation, lipid peroxidation, MMP decline, and cytochrome c expulsion which results in apoptosis signaling and cell loss.     

Ameliorated effects of garlic (Allium sativum) on biomarkers of subchronic acrylamide hepatotoxicity and brain toxicity in rats

Acrylamide (ACR) exerts its toxicity through stimulation of the oxidative stress; yet, its effect on neurotransmitter catabolic enzymes has not been elucidated. We investigated the effects of ACR exposure on brain and hepatic tissues antioxidant enzymes activities and different markers such as, acetylcholinesterase (AChE), nitric oxide (NO), monoamine oxidase (MAO), and lipid profile, and to evaluate the protective effects of garlic against ACR toxicity. Male Sprague-Dawley rats were exposed to ACR (1 mg kg^?1 body weight) with or without diet containing 1.5% of garlic powder for 40 days. ACR administration showed a decrease in AChE activity associated with an increase in MAO activity in both brain and hepatic tissues. In addition, ACR administration increased the lipid peroxidation and NO levels of both tissues while decreased the activities of glutathione (GSH), superoxide dismutase, and glutathione-S-transferase (GST). On the other hand, the activities of glutathione peroxidase (GPx) and catalase activities increased as a consequence of GSH depletion after ACR exposure. Finally, ACR exposure increased the brain and liver lipid profile of cholesterol, triglycerides and total lipid, while phospholipids level was decreased. Coadministration of garlic powder with ACR significantly attenuated oxidative stress, MAO activity, and inflammation in brain and hepatic tissues but did not ameliorate AChE activity. In conclusion, our results emphasized the role of garlic as a potential adjuvant therapy to prevent ACR neurotoxicity and hepatotoxicity.     

Vitamins E and C reduce oxidative stress and mitochondrial permeability transition caused by camptothecin – an in vitro study

Camptothecin (CPT), a broad spectrum antineoplastic agent, is known to induce oxidative stress and mitochondria are among the main sources of intracellular reactive oxygen species (ROS). We investigated the merit of vitamins E and C supplementation on CPT-induced mitochondrial alterations in vitro. Following treatment of isolated liver mitochondria with CPT, we assessed the mitochondrial membrane permeability transition (MPT), concentration of malondialdehyde, antioxidants and activities of the enzymes of the respiratory chain and Krebs cycle. Our results provide evidence that CPT caused mitochondrial swelling, increased lipid peroxidation and transition of mitochondrial permeability. The CPT lowered the levels of reduced mitochondrial thiols suggesting that thiol oxidation is the mechanism underlying CPT-induced MPT. Identical experiments were also performed after preincubating the mitochondria with vitamins E and C. It was found that vitamins E and C pretreatment inhibited the deleterious effects of CPT and loss of enzyme activity was restored by antioxidant supplementation. Our results suggest that the toxicity of CPT was mediated by an increase in ROS production by mitochondria. However, the addition of vitamins E or C ameliorated the oxidative stress. We propose that an attempt to counteract the deleterious consequences of chemotherapy with nutritional therapies may be a rational approach in superior patient care especially in a disease like cancer.     

Oxidative stress-mediated cardiac mast cell degranulation

Pulmonary mast cell degranulation is a well-characterized response to diesel exhaust exposure. A primary constituent of fossil fuel combustion is sulfur dioxide (SO₂). SO₂ was shown to induce mast cell degranulation in an immortalized cell line secondary to induction of intracellular oxidative stress; however, it is not known whether SO₂-induced oxidative stress directly triggers the activation of cardiac mast cells. Accordingly, this study sought to determine whether Na₂SO₃ induces degranulation of cardiac mast cells, and furthermore whether cardiac mast cell activation may be prevented by inhibition of oxidative stress. To this end, cardiac mast cells were isolated from epicardial surface of the heart and incubated with increasing concentrations of Na₂SO₃ (0, 0.5, or 5 mM). Antioxidant compounds targeting different mechanisms of free radical generation, including ebselen, diphenyleneiodonium (DPI), or α-tocopherol, were incubated with 5 mM of Na₂SO₃ in order to determine their efficacy in preventing mast cell degranulation. Na₂SO₃ induced a significant concentration-dependent histamine release from cardiac mast cells ranging from 8.87% to 18.86%, which was prevented by ebselen. No effect was observed with DPI and α-tocopherol under these conditions. In conclusion, these findings indicate that SO₂ is capable of producing cardiac mast cell degranulation in vitro; however, the variable effectiveness of the three antioxidants evaluated is indicative of a multifactorial mechanism mediating SO₂-induced cardiac mast cell degranulation. The particular effectiveness of ebselen in preventing mast cell degranulation may be related to its multiple mechanisms of preventing oxidative stress.     

Mercuric chloride effects on adult rat oval cells-induced apoptosis

In the present investigation, the toxicity of mercuric chloride (HgCl₂) was evaluated in adult oval cells isolated from rat utilizing the 2-acetylaminofluorene/partial hepatectomy technique. Isolated oval cells were incubated with 5 μM of HgCl₂ for 8 hr to elucidate in vitro cytotoxic responses. Recently, autophagic cell death was found in rat hepatocytes in vitro within 30 min of incubation with 5 μM of mercury (Hg) which triggered apoptosis and necroptosis in a time-dependent manner. Nuclear degradation occurred within 30 min of incubation and progressed with time until 8 hr. Apoptosis evidenced by activation of caspase-dependent pathway between 30 min to 8 hr of incubation was mediated via interchange of death domain signaling pathways. Receptor-interacting protein played a positive role to modulate the death domain receptors in the scenario of apoptotic death of oval cells until 6 hr. Autophagic marker proteins ATG12 and LC3B exerted a significant role in triggering apoptosis in 5 μM Hg-treated oval cells. No apparent expression of apoptosis-inducing-factor (AIF) and HMGB1 indicated absence of caspase-independent apoptosis and necrosis in the rat oval cells between 30 min and 8 hr. Thus a low concentration of Hg modulates programmed cell death in adult rat oval cells by altering expression of proteins involved in the molecular mechanisms of cellular functions.     

Cadmium sulfide nanoparticle induces oxidative stress and pro-inflammatory effects in human lung adenocarcinoma epithelial cells

Cadmium sulfide nanoparticles (CdSNP) are increasingly used in biological applications. This study was undertaken to understand the mechanisms underlying adverse effects of CdSNP using human lung adenocarcinoma epithelial (A549) cells. Cellular toxicity was evaluated by using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide and neutral red assays. Results showed that CdSNP reduced mitochondrial function and induced lysosomal activity in concentration and time-dependent manner. CdSNP produced oxidative stress as evidenced by reduction of glutathione (GSH) levels and increase in reactive oxygen species and lipid peroxidation levels. Induction of caspase-3 enzymes and condensed, fragmented nuclei was observed in CdSNP-treated cells. Furthermore, the levels of interleukin-8, tumor growth factor and DNA fragmentation were significantly higher in CdSNP exposed cells. Data indicated that toxicity of CdSNP noted in A549 cells may be mediated through oxidative stress. This study warrants more comprehensive assessment of CdSNP prior to industrial applications.     

Carbon black-benzo(a)pyrene complex-induced apoptosis and autophagy blockage in rat alveolar macrophages

Abstract

Cooking of foods and the burning of biomass and fossil fuels in stoves are the main sources of cooking fumes, with carbon black and polycyclic aromatic hydrocarbons as main components. The toxicity of carbon black and polycyclic aromatic hydrocarbons has been well studied individually, but the combined toxicity is much less understood. Carbon black can adsorb benzo(a)pyrene to form a complex which displays an altered physical form which in this study has been constructed to simulate particles in the cooking fumes and explore the combined toxic effect on rat alveolar macrophages. The complex-induced cell apoptosis and blocked cell autophagy flux compared with both individually. The mechanism of toxicity may be by intracellular reactive oxygen species generation, impairing the mitochondrial membrane potential and activating apoptosis signaling pathways.     

Oxidative stress mediated cytotoxicity of TiO2 nano anatase in liver and kidney of Wistar rat

This study examined the adverse effects of TiO₂ nanoparticle (nano-TiO₂) on the kidney and liver of Wistar rats. Changes of serum biochemical parameters and pathological lesions indicated that liver and kidney were significantly affected in animals treated with 50?mg?kg^?1 of nano-TiO₂. The inverse relationship between the level of reactive oxygen species and the activities of superoxide dismutase, catalase, and glutathione peroxidase indicates that nano-TiO₂ induces oxidative stress. A significant increase in the apoptosis of liver and kidney in a dose-dependent manner was also observed. The ultrastructural observations confirmed the internalization of nano-TiO₂ and their direct involvement in the mitochondria-mediated cytotoxicity. Data indicated that nano-TiO₂ induce oxidative stress which produces genotoxicity such as oxidative DNA damage, micronuclei (MN) induction, and cell apoptosis in liver and kidney.     

Studies on mercuric sulfide,a component of cinnabar,a Chinese herbal medicine,on celluar functions in mouse lung and fibroblasts

Although the cytotoxic effects of mercuric chloride (HgCl₂) and methylmercury chloride (MeHg) have been extensively studied, the insoluble mercuric sulfide (HgS) has been the subject of fewer studies. Since the traditional Chinese mineral drug, cinnabar (containing >95% HgS) continues to be used as an ingredient for infant sedation, the pharmacological and toxicological effects of HgS need to be clarified. In previous experiments, HgS and cinnabar were shown to be absorbed from the gastrointestinal tract (GIT) and distributed in various tissues including the lungs. Thus, a preliminarily examination of whether HgS might exert any oxidative stress on a mouse lung was undertaken. HgS reduced GSH content and increased lipid peroxidation in the lung. Further studies on the cytotoxic effects and the possible mechanisms of action of HgS were compared with HgCl₂ and MeHg in cultured lung fibroblast V₇₉ cells. The results showed that HgS produced cytotoxicity at a concentration (400–1200 µM)in a dependent manner with IC₅₀ of 795.6 µM, as compared to HgCl₂ and MeHg, 8.1 µM and 5.9 µM, respectively. In addition, the HgS induced the phenomena of DNA fragmentation, increasing reactive oxygen species (ROS) and decreasing mitochondrial membrane potential, accompanied by decreased levels of intracellular ATP and GSH and higher lipid peroxidation levels, similar to HgCl₂ and MeHg, but with different toxicokinetic properties. These findings provide evidence for understanding the mechanisms underlying the toxic effects of HgS.     

活性氧和钙信号参与铅对酵母细胞的毒性作用

以模式生物酵母菌为材料,研究铅对细胞的毒性效应,探讨胞内活性氧(ROS)和Ca~(2+)在铅诱导细胞死亡中的作用。结果显示,浓度为5~100 mg·L~(-1)的硝酸铅可降低酵母细胞活性,诱导酵母细胞死亡,随着铅浓度的提高和作用时间的延长,细胞死亡率增高。在铅处理组酵母细胞中,ROS和Ca~(2+)水平显著升高,线粒体膜电位明显下降;用1 mmol·L~(-1)的外源抗坏血酸(AsA)能降低铅引发的酵母细胞死亡,0.5 mmol·L~(-1)的钙离子螯合剂乙二醇双四乙酸(EGTA)或0.1 mmol·L~(-1)的质膜Ca~(2+)通道特异性抑制剂氯化镧(LaCl_3)亦可明显抑制铅引起的酵母细胞死亡。研究结果表明,铅诱发的酵母细胞死亡与处理组胞内ROS和Ca~(2+)升高有关,高浓度的Ca~(2+)可能通过诱导线粒体膜通透性转变孔道开放,或者高水平ROS可能损伤线粒体膜,致线粒体膜电位下降,继而激活相关下游信号导致细胞死亡。