Potentiated Interaction between Ineffective Doses of Budesonide and Formoterol to Control the Inhaled Cadmium-Induced Up-Regulation of Metalloproteinases and Acute Pulmonary Inflammation in Rats

The anti-inflammatory properties of glucocorticoids are well known but their protective effects exerted with a low potency against heavy metals-induced pulmonary inflammation remain unclear. In this study, a model of acute pulmonary inflammation induced by a single inhalation of cadmium in male Sprague-Dawley rats was used to investigate whether formoterol can improve the anti-inflammatory effects of budesonide. The cadmium-related inflammatory responses, including matrix metalloproteinase-9 (MMP-9) activity, were evaluated. Compared to the values obtained in rats exposed to cadmium, pretreatment of inhaled budesonide (0.5 mg/15 ml) elicited a significant decrease in total cell and neutrophil counts in bronchoalveolar lavage fluid (BALF) associated with a significant reduction of MMP-9 activity which was highly correlated with the number of inflammatory cells in BALF. Additionally, cadmium-induced lung injuries characterized by inflammatory cell infiltration within alveoli and the interstitium were attenuated by the pre-treatment of budesonide. Though the low concentration of budesonide (0.25 mg/15 ml) exerted a very limited inhibitory effects in the present rat model, its combination with an inefficient concentration of formoterol (0.5 mg/30 ml) showed an enhanced inhibitory effect on neutrophil and total cell counts as well as on the histological lung injuries associated with a potentiation of inhibition on the MMP-9 activity. In conclusion, high concentration of budesonide alone could partially protect the lungs against cadmium exposure induced-acute neutrophilic pulmonary inflammation via the inhibition of MMP-9 activity. The combination with formoterol could enhance the protective effects of both drugs, suggesting a new therapeutic strategy for the treatment of heavy metals-induced lung diseases.     

Cadmium carcinogenesis

Cadmium is a heavy metal of considerable environmental and occupational concern. Cadmium compounds are classified as human carcinogens by several regulatory agencies. The most convincing data that cadmium is carcinogenic in humans comes from studies indicating occupational cadmium exposure is associated with lung cancer. Cadmium exposure has also been linked to human prostate and renal cancer, although this linkage is weaker than for lung cancer. Other target sites of cadmium carcinogenesis in humans, such as liver, pancreas and stomach, are considered equivocal. In animals, cadmium effectively induces cancers at multiple sites and by various routes. Cadmium inhalation in rats induces pulmonary adenocarcinomas, in accord with its role in human lung cancer. Cadmium can induce tumors and/or preneoplastic lesions within the rat prostate after ingestion or injection. At relatively high doses, cadmium induces benign testicular tumors in rats, but these appear to be due to early toxic lesions and loss of testicular function, rather than from a specific carcinogenic effect of cadmium. Like many other metals, cadmium salts will induce mesenchymal tumors at the site of subcutaneous (s.c.) or intramuscular (i.m.) injections, but the human relevance of these is dubious. Other targets of cadmium in rodents include the liver, adrenal, pancreas, pituitary, and hematopoietic system. With the exception of testicular tumors in rodents, the mechanisms of cadmium carcinogenesis are poorly defined. Cadmium can cause any number of molecular lesions that would be relevant to oncogenesis in various cellular model systems. Most studies indicate cadmium is poorly mutagenic and probably acts through indirect or epigenetic mechanisms, potentially including aberrant activation of oncogenes and suppression of apoptosis.     

Study of the effect of the administration of Cd(II), cysteine, methionine, and Cd(II) together with cysteine or methionine on the conversion of xanthine dehydrogenase into xanthine oxidase

Cadmium is known as to be a potent pulmonary carcinogen to human beings and to induce prostate tumor. The sequestration of cadmium, an extremely toxic element to living cells, which is performed by biological ligands such as amino acids, peptides, proteins or enzymes is important to minimize its participation in such deleterious processes. The synthesis of metallothionein is induced by a wide range of metals, in which cadmium is a particularly potent inducer. This protein is usually associated with cadmium exposure in man. Because metallothioneins may act as a detoxification agent for cadmium and chelation involves sulfur donor atoms, we administered only cadmium, cysteine, or methionine to rats and also each of these S-amino acids together with cadmium and measured the production of superoxide radicals derived from the conversion of xanthine dehydrogenase to xanthine oxidase. It could be seen in this work that the presence of cadmium enhances this conversion. However, its inoculation with cysteine or methionine almost completely diminishes this effect and this can be the result of the fact that these amino acids complex Cd(II). Thus, these compounds can be a model of the action of metallothionein, removing cadmium from circulation and preventing its deleterious effect.     

Effects of essential divalent metals on carcinogenicity and metabolism of nickel and cadmium

Interactions between the physiologically essential metals calcium, magnesium, and zinc and the carcinogenic metals nickel and cadmium were investigated to help elucidate the mechanisms of action of the carcinogenic metals. Bioassay studies revealed several significant findings, including: (1) the ability of magnesium and calcium to inhibit nickel-induced elevation of pulmonary adenoma incidence in strain A mice; (2) the ability of magnesium, but not of calcium, to prevent cadmium-induced subcutaneous sarcoma formation; and (3) the ability of magnesium, but not of calcium, to inhibit nickel-induced muscle tumor formation. Biochemical studies indicated a direct relationship between the antitumorigenic potential of magnesium and the capacity of this metal to: (1) inhibit nickel and cadmium uptake by the target tissues in vivo; (2) inhibit nickel-induced disturbances in DNA synthesis in vivo; (3) inhibit nuclear and cytosolic uptake of nickel by the target tissue cells in vivo; and (4) inhibit nickel and cadmium binding to DNA in vitro. Calcium, which in most cases did not prevent carcinogenesis, had no consistent influence on the uptake of carcinogenic metals or their biochemical effects in the target tissues. Magnesium and zinc, but not calcium, were also found to attenuate the acute toxic effects of nickel, indicating a possible correlation between prevention of acute effects and reduction in tumorigenicity. Zinc, which antagonizes cadmium tumorigenicity in the rat testis, was found to reduce markedly cadmium uptake into isolated testicular interstitial cells. Also, zinc was found to inhibit strongly cadmium binding to DNA in vitro.     

Cadmium carcinogenesis in review

Cadmium is an inorganic toxicant of great environmental and occupational concern which was classified as a human carcinogen in 1993. Occupational cadmium exposure is associated with lung cancer in humans. Cadmium exposure has also, on occasion, been linked to human prostate cancer. The epidemiological data linking cadmium and pulmonary cancer are much stronger than for prostatic cancer. Other target sites for cadmium carcinogenesis in humans (liver, kidney, stomach) are considered equivocal. In rodents, cadmium causes tumors at several sites and by various routes. Cadmium inhalation in rats results in pulmonary adenocarcinomas, supporting a role in human lung cancer. Prostate tumors and preneoplastic proliferative lesions can be induced in rats after cadmium ingestion or injection. Prostatic carcinogenesis in rats occurs only at cadmium doses below those that induce chronic degeneration and dysfunction of the testes, a well-known effect of cadmium, confirming the androgen dependency of prostate tumors. Other targets of cadmium in rodents include the testes, adrenals, injection sites, and hematopoietic system. Various treatments can modify cadmium carcinogenesis including supplemental zinc, which prevents cadmium-induced injection site and testicular tumors while facilitating prostatic tumors. Cadmium is poorly mutagenic and probably acts through indirect mechanisms, although the precise mechanisms remain unknown.     

Cadmium accumulation and interactions with zinc, copper, and manganese, analysed by ICP-MS in a long-term Caco-2 TC7 cell model

The influence of long-term exposure to cadmium (Cd) on essential minerals was investigated using a Caco-2 TC7 cells and a multi-analytical tool: microwave digestion and inductively coupled plasma mass spectrometry. Intracellular levels, effects on cadmium accumulation, distribution, and reference concentration ranges of the following elements were determined: Na, Mg, Ca, Cr, Fe, Mn, Co, Ni, Cu, Zn, Mo, and Cd. Results showed that Caco-2 TC7 cells incubated long-term with cadmium concentrations ranging from 0 to 10 μmol Cd/l for 5 weeks exhibited a significant increase in cadmium accumulation. Furthermore, this accumulation was more marked in cells exposed long-term to cadmium compared with controls, and that this exposure resulted in a significant accumulation of copper and zinc but not of the other elements measured. Interactions of Cd with three elements: zinc, copper, and manganese were particularly studied. Exposed to 30 μmol/l of the element, manganese showed the highest inhibition and copper the lowest on cadmium intracellular accumulation but Zn, Cu, and Mn behave differently in terms of their mutual competition with Cd. Indeed, increasing cadmium in the culture medium resulted in a gradual and significant increase in the accumulation of zinc. There was a significant decrease in manganese from 5 μmol Cd/l exposure, and no variation was observed with copper.     

Cadmium Stress Disrupts the Endomembrane Organelles and Endocytosis during Picea wilsonii Pollen Germination and Tube Growth

As one of the most severe pollutants, cadmium has been reported to be harmful to plant cells, but the effects of cadmium on gymnosperm pollen germination and tube growth and the mechanism of this involvement are still unclear. Here, we report that cadmium not only strongly inhibited P. wilsonii pollen germination and tube growth, but also significantly altered tube morphology in a dose-dependent manner. Time-lapse images obtained with a laser scanning confocal microscope revealed that endocytosis was dramatically inhibited by cadmium stress. Further investigation with ER-Tracker dye indicated that cadmium stress reduced the number of the Golgi apparatus, and induced dilation of ER. Additionally, Lyso-Tracker staining showed that cadmium distinctly promoted the formation of acidic organelles in pollen tubes, likely derived from the dilated ER. Taken together, our studies indicated that P. wilsonii pollens were highly susceptible to cadmium stress, and that cadmium stress strongly inhibited pollen germination and tube growth by disrupting the endomembrane organelles, inhibiting endo/exocytosis, and forming acidic vacuoles, resulting in swollen tube tips and irregularly broadened tube diameters. These findings provide a new insight into the effects of cadmium toxicity on the tip growth of pollen tubes.     

Immunotoxicity of soluble and insoluble salts of cadmium instilled intratracheally

Rats were intratracheally (i.t.) exposed to 36.5 or 27.5 microg of cadmium (Cd) as soluble cadmium chloride (CdCl2) and insoluble cadmium oxide (CdO) salts. The retention of metal in lungs, liver and kidney was assessed by atomic adsorption spectrophotometer. The animals were intraperitoneally (i.p.) primed with sheep red blood cells (SRBC) and assessed for the number of antibody forming cells in lung associated lymph nodes (LALN) and spleen. Both the compounds had similar retention of metal in lungs but CdO induced more pulmonary inflammatory and degradative changes than CdCl2. The larger influx of polymorphonuclear cells (PMNs) following CdO exposure appears to be due to the absence of protection afforded by Cd induced metallothionein cytoplasmic protein while the Cd metallothionein complex formed in the case of CdCl2 is more protective. However both forms of Cd had similar local immunosuppressive potential but CdO had more prolonged suppressive effect.     

The tissue disposition and urinary excretion of cadmium, zinc, copper and iron, following repeated parenteral administration of cadmium to rats.

The effect of repeated parenteral administration of cadmium (0.75, 1.5 and 3.0 mg/kg) on tissue disposition and urinary excretion of cadmium, zinc, copper and iron has been studied in the male rat. Cadmium, zinc and copper accumulated in liver and kidney, but the concentration of iron did not alter significantly. The kidney weight relative to body weight showed a dose-related increase in weight of 25--65%. Excretion of cadmium in the urine increased directly with dosage and the increase was most significant when kidney damage had probably occurred. Administration of cadmium also resulted in dose-related increases in the urinary excretion of zinc, copper and iron. The cadmium concentration of blood increased with dosage of cadmium, and the plasma concentrations of zinc and copper were also raised but plasma iron concentration was diminished.     

Leaf-targeted phytochelatin synthase in Arabidopsis thaliana

One of the key steps in developing transgenic plants for the phytoremediation of metal containing soils is to develop plants that accumulate metals in the aerial tissues. With the goal of changing the distribution of phytochelatin (PC)-dependent cadmium accumulation from roots to the leaves, the phytochelatin synthase (PCS) deficient cad1-3 mutant and wild type (Col-0) Arabidopsis plants were transformed with an Arabidopsis phytochelatin synthase (AtPCS1) under the control of a leaf-specific promoter. Three independent transformant lines from each genetic background were chosen for further analysis and designated cad-PCS and WT-PCS. PCS activity in the cadPCS lines was restored in the leaves, but not in the roots. Additionally, when whole plants were treated with cadmium, PCs were found only in the leaves of cad-PCS plants. Although the inserted AtPCS1 gene was leaf-specific, cad-PCS lines showed an overall decrease in cadmium toxicity evidenced by a partial amelioration of the "brown-root" phenotype and root growth was restored to wild type levels when treated with cadmium and arsenate. WT-PCS lines showed an increase in leaf PCS activity but had only wild type PC levels. In addition, cadmium uptake studies indicated that there was no difference in cadmium accumulation among all types tested. So, while we were able to protect the plants against cadmium by expressing PC synthase only in the leaves, we were not able to limit cadmium accumulation to aerial tissues.     

The effects of heavy metals on common carp white blood cells in vitro

The in vitro effects of cadmium, copper, lead and zinc, and various cadmium compounds (chloride, sulphate and nitrate) on common carp (Cyprinus carpio) lymphocyte viability and phagocyte activity, were evaluated. The percentage of dead lymphocytes was determined after Trypan blue staining, and phagocyte activity was measured by using the nitroblue tetrazolium (NBT) reduction test. Lead was the most toxic to lymphocytes--the maximum mortality exceeded 30%, and was significantly higher at 1 microM of lead, compared to the control. The maximum mortality caused by cadmium was below 10%, but was significantly elevated with 5 microM or more of cadmium. Zinc induced lymphocyte mortality from 10 microM, whilst no effect was observed with copper. The incubation of full blood with the three cadmium compounds (at 5mg/l of cadmium) showed that cadmium nitrate and cadmium sulphate were more toxic (over 35% and 25% mortality, respectively) than cadmium chloride (about 15% mortality). This was confirmed by the results of tests on isolated cells--1mg/l of cadmium as nitrate and sulphate increased lymphocyte mortality compared to the control and cadmium chloride. Phagocytic activity was less sensitive to heavy metals than was lymphocyte viability. It was significantly reduced following exposure to 50 microM and 100 microM cadmium, and 100 microM zinc, but no effects were observed with either lead or copper.     

Transgenic tobacco plants expressing a rice cysteine synthase gene are tolerant to toxic levels of cadmium

Plants tolerate heavy metals through sequestration with cysteine-rich peptides, phytochelatins. In this reaction, the rate limiting step is considered to be the supply of cysteine, which is synthesized by cysteine synthase (CS, EC 4.2.99.8) from hydrogen sulfide andO-acetylserine. In this study, we transformed tobacco (Nicotiana tabacum) plants withRCS1, a cytosolic cysteine synthase gene of rice (Oryza sativa), and examined their sensitivity to cadmium. The transgenic plants had up to 3-fold higher activity of cysteine synthase than wild-type plants. Upon exposure to cadmium, they exhibited obvious tolerance with much greater growth than wild-type plants. The level of phytochelatins in shoots was higher in transgenic than in wild-type plants after cadmium treatment, suggesting that cadmium was actively trapped by phytochelatins. However, the cadmium concentration per g fresh weight of whole transgenic plants was 20 percnt; lower than that of wild-type plants, suggesting cadmium to be either actively excreted or diluted by fast growth. Genetic analysis of progenies clearly showed segregation of cadmium tolerance, indicating that the trait resulted from the introduced gene. These results suggest that introduction of a cysteine synthase gene into tobacco plants resulted not only in high level production of sulfur-containing compounds that detoxify cadmium, but also in active elimination of cadmium toxicity from plant bodies.     

镉胁迫对草莓光合的影响

通过盆栽试验,研究了Cd胁迫对草莓叶片叶绿素含量、光合速率、气孔导度、蒸腾速率和胞间二氧化碳浓度的影响.结果表明,Cd降低草莓叶片中的叶绿素总量,并改变叶绿素a和叶绿素b的比例;Cd降低草莓叶的光合速率和气孔导度,但在Cd胁迫初期,低浓度的Cd对草莓叶的光合速率和气孔导度有促进作用;Cd还降低草莓叶片的蒸腾速率和胞间CO₂浓度,其中,胞间CO₂浓度的降低幅度相对较小.     

镉胁迫对姬松茸生长和镉吸收累积的影响

利用外源镉添加的袋栽试验,系统研究了镉对姬松茸J37和J1两菌株农艺性状、子实体产量、氨基酸和镉含量的影响,以求为低富集镉的姬松茸品种选育和安全栽培提供科学依据。结果表明外源镉添加对姬松茸J37和J1品种子实体的农艺性状的影响各异,两菌株单个子实体重量、子实体高度、菌盖直径、菌盖厚度和菌柄直径等均与外源镉浓度具有显著的负相关性,其中子实体重量与外源镉浓度间的相关性最大;15 mg/kg镉水平是两菌株农艺性状发生显著变化的敏感点。不同镉水平下J37菌株单个子实体重量比J1菌株高10.9%—36.6%,表现为较强镉耐受性。姬松茸J37与J1菌株子实体产量、氨基酸和镉含量均随着潮次的增加而降低,子实体产量和氨基酸含量随着外源镉水平增加而降低,而镉含量随外源镉水平增加而增加。与外源镉0mg/kg处理相比,当外源镉水平达15 mg/kg时,J1和J37子实体总产量分别下降23.4%和13.4%以上,J1和J37子实体镉含量分别提高51.5%、13.7%以上,且J1子实体镉含量已达到食品卫生标准的临界值。当外源镉水平达到35 mg/kg时,J1和J37子实体产量分别下降45%、32%以上,J1和J...     

Age characteristics of cadmium content in the organism of poisoned rats with experimental metabolic acidosis

The paper demonstrates age peculiarities of cadmium content in the liver, kidneys and spleen of 3-month and 18-month rats poisoned by cadmium sulfate. It was registered that the content of cadmium is 266.7 times higher in 3-month old rats, and 141.8 times in 18 months rats in comparison with intact ones. A biological model of introducing 3-month and 18-month rats into the state of metabolic acidosis before and after poisoning with cadmium sulfate was developed. The result of the research showed that changes in acid-base equilibrium of poisoned rats towards metabolic acidosis may influence a decrease of cadmium content in their organisms. Thus, when modeling of metabolic acidosis before the poisoning with cadmium, the content of cadmium decreases in 3-month rats' liver by 21%, in kidneys by 53%; in metabolic acidosis modeling after cadmium poisoning its content decreases, accordingly, by 44% and by 56.5% in comparison with the just poisoned ones. In metabolic experimental acidosis modeling the content of cadmium in 3-month poisoned rats' spleen decreased by 36.5% before and after poisoning. Such changes were also registered in 18-month old rats but to lower extent. Thus, the results of the researches showed more effective correction of cadmium intoxication decrease in the organism of 3-month rats when using the model of acid-base equilibrium change in the organisms of poisoned animals.     

Role of respiration and glutathione in cadmium-induced oxidative stress in Escherichia coli K-12

Cadmium is a widespread pollutant that has been associated with oxidative stress, but the mechanism behind this effect in prokaryotes is still unclear. In this work, we exposed two glutathione deficient mutants (ΔgshA and ΔgshB) and one respiration deficient mutant (ΔubiE) to a sublethal concentration of cadmium. The glutathione mutants show a similar increase in reactive oxygen species as the wild type. Experiments performed using the ΔubiE strain showed that this mutant is more resistant to cadmium ions and that Cd-induced reactive oxygen species levels were not altered. In the light of these facts, we conclude that the interference of cadmium with the respiratory chain is the cause of the oxidative stress induced by this metal and that, contrary to previously proposed models, the reactive oxygen species increase is not due to glutathione depletion, although this peptide is crucial for cadmium detoxification.     

Early reactions of cadmium with Ehrlich cells.

Ehrlich ascites tumor cells accumulate cadmium against a concentration gradient in a bisphasic uptake process. There is little efflux of the metal from preloaded cells into a cadmium-free medium. Incorporation of 3H-thymidine into DNA is markedly inhibited by cadmium ion at 5-100 ng atoms of Cd/mg of cell protein, but uptake of the nucleoside label into cells is not depressed in this concentration range. Cell respiration is much less affected by cadmium ion despite the sensitivity of isolated mitochondria to the metal. Model experiments using several cadmium complexes with known conditional formation constants show that bovine heart mitochondria have strong affinity for cadmium ion. The contrast between this result and the resistance of cells to respiratory inhibition with cadmium ion is discussed to illustrate the difficulty in relating in vitro studies to the cell. The behavior of cadmium ion with the Ehrlich cell is compared with data for zinc ion to reveal similarities in inhibition of nucleoside metabolism and respiration but a sharp difference in transport properties.     

南瓜对镉的吸收积累特性研究

对南瓜进行盆栽试验,通过加入不同含量的镉、CaCO₃和草炭土,研究南瓜对镉的吸收积累特性,利用火焰原子吸收法测定。结果表明,南瓜植株对镉的富集量主要集中在茎、根中;当土壤镉含量小于5mg·kg^-1时,对南瓜根的生长有促进作用,当土壤镉含量大于5mg·kg^-1时,开始对根的生长产生抑制作用,抑制作用随镉含量的增大而加强;在土壤中施CaCO₃,能降低南瓜对镉的吸收;在土壤中加入草炭土,在一定范围内,它能促进南瓜对镉的吸收。     

镉在大鼠前列腺组织内的蓄积

为比较重金属镉在去势与非去势雄性大鼠前列腺组织内蓄积量的变化,将60只健康雄性SD大鼠随机分为六组:对照组、去势对照组、低剂量镉组(2.5 mμmol·Kg^-1)、高剂量镉组(20 mμmol·Kg^-1)、去势低剂量镉(2.5 mμmol·Kg^-1)组、去势高剂量镉(20 mμmol·Kg^-1)组,18个月后取材进行实验.利用原子吸收分光光度法检测大鼠前列腺组织中重金属镉含量.与非去势对照组相比,各去势组大鼠前列腺组织内镉含量明显降低;注射镉后,大鼠前列腺组织镉含量明显高于对照组,但高、低剂量组之间无显著差别.实验表明长期镉暴露后,前列腺组织内镉含量明显增加;经去势手术的大鼠,长期镉暴露后前列腺组织内镉含量未升高,但机制并不明确.     

Impact of Soil Cadmium on Land Snails: A Two-Stage Exposure Approach under Semi-Field Conditions Using Bioaccumulative and Conchological End-Points of Exposure

Land snails are highly tolerant to cadmium exposure and are able to accumulate soil cadmium independently of food ingestion. However, little information exists on the kinetics of cadmium retention in terrestrial gastropods exposed to an increase in the soil cadmium content, over time. There is also little knowledge about how exposure to cadmium-polluted soils influences shell growth and architecture. In this context, we examined cadmium accumulation in the hepatopancreas and shell of juvenile Cantareus aspersus exposed to elevating high levels of cadmium in soil. Also, the toxicity of cadmium to snails was assessed using a range of conchological endpoints, including shell height, width, volume, allometry and integrity. Test snails, aged three months, were reared under semi-field conditions, fed an uncontaminated diet and exposed first, for a period of 30 days, to a series of soil cadmium concentrations, and then, for a second period of 30 days, to soils with higher cadmium content. Cadmium showed a dose-dependent accumulation in both the hepatopancreas and shell. The kinetics of cadmium retention in the hepatopancreas of snails previously exposed to cadmium-spiked soils was significantly influenced by a new exposure event. The shell was not a relevant bioaccumulator for soil cadmium. Under the present experimental conditions, only high cadmium exposure significantly affected either the shell growth or snail survival. There was no consistent effect on shell allometry, but the shell integrity, especially in rapidly growing parts, appeared to be affected by high cadmium exposure. Our results attest to the value of hepatopancreas for describing cadmium retention in land snails and to the difficulty of using conchological parameters in field surveys for estimating the environmental hazard of soil cadmium.