Cadmium/metabolism* ; Kidney ; Cells, Cultured ; Mitochondria ; Metallothionein/metabolism* ; Liver

To investigate the effects of metallothionein (MT) on isolated rat heart, 16 Wistar rats were randomly divided into 2 groups. In control group (group C), distilled water was injected intraperitoneally and 24 h later isolated hearts were perfused with Langendorff and stored at 4 degrees C for 3 h with histidine-tryptophan-ketoglutarate (HTK) solutions, and then isolated hearts were perfused for 2 h by Langendorff. In experimental group (group E), 3.6% ZnSO(4) was injected intraperitoneally, 24 h later isolated hearts were perfused by Langendorff and stored at 4 degrees C for 3 h with HTK solutions, and then the isolated hearts were perfused for 2 h with Langendorff. MT content, the recovery of hemodynamics, myocardial water content (MWC), lactate dehydrogenase (LDH) and creatine kinase (CK) leakage, adenosine triphosphate (ATP) and malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, myocardial cell Ca(2+) content, Ca(2+)-ATPase activity of mitochondria ([Ca(2+)-ATPase](m)) and its Ca(2+) content ([Ca(2+)](m)), synthesizing ATP activity of mitochondria ([ATP](m)), and the ultrastructure of cells were examined. There were a significant increase in group E in hemodynamic recovery, ATP content, SOD activity, [Ca(2+)-ATPase](m) activity, [ATP](m) activity, and substantial reduction in MWC, LDH and CK leakage, MDA content, myocardial cell Ca(2+) content, [Ca(2+)](m) content, and the ultrastructural injury were obviously milder than that of group C. This study demonstrated that MT has protective effects on isolated rat heart.
Cardiotonic Agents/*pharmacology ; Creatine Kinase/*metabolism ; L-Lactate Dehydrogenase/metabolism ; Metallothionein/biosynthesis ; Metallothionein/*pharmacology ; Myocardium/*metabolism ; Myocardium/ultrastructure ; Random Allocation ; Rats, Wistar ; Superoxide Dismutase/metabolism ; Zinc Sulfate/pharmacology

Metallothionein (MT) family, intracellular zinc binding proteins, has been suggested to play an important regulatory role in zinc metabolism. The present study utilized light and confocal microscopic methods to investigate the expression pattern of MT-1, 2 and to determine the distribution and extent of colocalization of MT-1, 2 relative to subpopulations of S100 positive folliculostellate (FS) cells and somatotrophs in normal rat anterior pituitary. By light microscopy, MT-1, 2 immunoreactivity was observed both in pars intermedia and pars distalis of anterior pituitary. Confocal microscopy revealed that most MT immunoreactivity was colocalized in S100 positive FS cells, not in somatotrophs. This is the first report that FS cells in pars distalis contain MT-1, 2. These results suggest that MT family may be involved in regulation of hypophyseal endocrine functions and can be used as new markers of FS cells.
Animals ; Carrier Proteins ; Humans ; Immunohistochemistry ; Metabolism ; Metallothionein ; Microscopy ; Microscopy, Confocal ; Pituitary Gland, Anterior* ; Rats* ; Somatotrophs ; Zinc

The effect of dexamethasone (10(-5)M) and epinephrine (10(-6)M) on the biosynthesis of metallothionein (MT) in the perfused rat liver was investigated. MT synthesis was determined by measuring the incorporation of 14C-L-aspartic acid into liver MT fraction after the perfusion for five hours of isolated liver by artificial blood containing 14C-L-U-aspartic acid (0.2uci) with dexamethasone or epinephrine. MT was isolated by Sephadex G-75 column chromatography and DEAE Sephadex column chromatography. Incorporation of radioactive 14C into the MT fraction of perfused liver cytosol (9.0grams of liver) from dexamethasone treated, epinephrine treated and control groups were, respective1y, 0.72, 0.34 and 0.33% of total radioactivity infused. Total protein content in the MT fraction of liver perfused with dexamethasone and epinephrine were 0.80, 0.64mg/g liver compared to 0.52mg/g liver in the control. MT, a protein having a high content of cystein and metals is synthesized in the perfused rat liver and its induction is stimulated by dexamethasone, while epinephrine increased the accumulation of Zn in the MT fraction of the perfused rat liver. The present experiment confirms that MT synthesis and degradation are somewhat regulated by glucocorticoid hormone and epinephrine.
Animal ; Dexamethasone/pharmacology* ; Epinephrine/pharmacology* ; Female ; In Vitro ; Liver/drug effects ; Liver/metabolism* ; Metalloproteins/metabolism* ; Metallothionein/metabolism* ; Perfusion ; Rats ; Zinc/metabolism

Metallothioneins (MTs) are a group of intracellular proteins of low molecular weight and rich cysteine, encoded by a family of genes containing at least 10 functional isoforms in human. MTs widely exist in the male reproductive system and are involved in many pathophysiological processes such as metal ion homeostasis and detoxification, protection against oxidative damage, cell proliferation and apoptosis, chemoresistance and radiotherapy resistance. This review describes the location and effects of MTs in the male reproductive system.
Gene Expression Profiling ; Genitalia, Male ; metabolism ; physiopathology ; Humans ; Male ; Metallothionein ; genetics ; metabolism ; physiology ; Protein Isoforms ; genetics ; metabolism ; physiology

AIMTo evaluate the effects of different doses of zinc on the expression of metallothionein isoforms in stressed hippocampal neurons in vitro.

METHODSThe cell stress model was developed by corticosterone. The cultured hippocampal neurons were assigned to seven groups as follows: control group, zinc deficiency group, and their corresponding stressed groups, as well as three different levels of zinc complementarity groups.

RESULTSIn zinc deficiency group, the expressions of metallothionein and MT-1 mRNA, MT-3 mRNA were downregulated. On the other hand, inductions of metallothionein and it's mRNAs in stressed zinc complementarity group were increased. In addition, the levels of supernatant IL-6 and NO were increased clearly in zinc deficiency group and corticosterone stressed groups.

CONCLUSIONOur results suggest that zinc deficiency may decrease while zinc complementarity increase the expressions of metallothioneins and MT-1 mRNA, MT-3 mRNA in stressed hippocampal neurons in vitro.

Animals ; Animals, Newborn ; Hippocampus ; cytology ; metabolism ; In Vitro Techniques ; Metallothionein ; metabolism ; Neurons ; metabolism ; RNA, Messenger ; Rats ; Zinc ; pharmacology

OBJECTIVETo explore the effects of cadmium on zinc metabolism and its function and the protective effects of pre-supplement zinc to it.

METHODSNS or different doses of CdCl(2) were injected to pregnant dams intraperitoneally at the 7th, 10th and 13th day of gestation respectively. At the 21st pregnant day embryos were taken out from the pregnant rats. Another rats of pre-supplement zinc or no pre-supplement zinc group were injected different doses of CdCl(2) or NS intraperitoneally after 6 days. After 24 hours the rats were killed. The contents of Cd, Zn and relative biomarkers of effect of liver, brain or serum were detected in both embryos and adult rats.

RESULTSCompared with control group, the contents of T-AOC and Ach were significantly reduced in the Cd treatment group in the embryonic brains, the activity of AKP in the embryonic liver tissues was decreased, and The Cd content was increased significantly in embryonic liver and was negatively correlated with the Zinc content in the embryonic brain. There were no differences in the activities of SOD and AKP and the contents of Cd and MDA between pre-supplement Zn control group and no supplement Zn control group, but higher content of Zn in liver and serum in the former. Compared with no supplement Zn control group, there were higher contents of Cd in liver and serum, Zn and MDA in liver, lower activities of SOD in liver and AKP in liver and serum, and lower content of Zn in serum in the Cd treatment groups. Pre-supplement Zn significantly increase the content of Zn and the activities of SOD in liver and AKP in serum, decrease the content of MDA in liver and Cd in serum resulted by Cd treatment only. The content of Zn and the activity of AKP in serum and the activities of SOD and AKP in liver were negatively correlated with the content of Cd in corresponding tissue significantly.

CONCLUSIONCadmium can enter embryo and enter brain by permeating the brain-blood barrier during the embryonic period. The decrease of AKP activity, some neural transmitter and capacity of anti-lipid peroxidation that are related with Zn in embryos are caused when the pregnant rats are administered with cadmium. Cd can inhibits the activities of AKP and SOD in liver, and the activity of AKP in serum respectively, and increase the content of MDA in liver dose-dependently. The effects induced by cadmium are related with zinc abnormal distribution. Pre-supplement zinc to rats can antagonize these effects in different degree.

Animals ; Cadmium ; toxicity ; Female ; Liver ; metabolism ; Male ; Maternal Exposure ; Metallothionein ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Zinc ; metabolism ; pharmacology

OBJECTIVETo screen and identify the protein interacting with HBV antigen in hepatocytes. Then investigate the biological functions of hepatitis B virus antigen in the pathogenesis of hepatitis B and seek effective methods to prevent and treat it.

METHODSThe yeast two-hybrid system-3 technique was used to construct HBV PreS2, HBeAg, HBcAg, HBxAg bait plasmids. The bait plasmids transformed the yeast AH109 and expressed themselves in it. After being identified by SDS-PAGE and Western blot, the AH109 yeast was mated with yeast Y187 containing liver cDNA library plasmid in 2 x YPDA medium to form diploid yeast and was then plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for screening. Plasmids of blue colonies were extracted and transformed into Escherichia coli, then analyzed by DNA sequencing and bioinformatics. To further prove the interaction between HBV antigen and metallothionein, translation was performed by using reticulocyte lysate and coimmunoprecipitation was displayed in vitro.

RESULTSGenes coding for HBV antigen binding protein were successfully cloned and metallothionein was found in that protein. The interaction between HBeAg, HBcAg and HBxAg and metallothionein were further proved by coimmunoprecipitation in vitro.

CONCLUSIONThe interaction between HBV antigen and metallothionein indicates that metallothionein may participate in the pathogenesis of hepatitis B

Hepatitis B ; metabolism ; Hepatitis B Antigens ; chemistry ; Hepatocytes ; metabolism ; Humans ; Metallothionein ; chemistry ; Protein Interaction Mapping ; Two-Hybrid System Techniques

Animals ; Calcium ; metabolism ; Copper ; metabolism ; Environmental Exposure ; prevention & control ; Erythrocytes ; metabolism ; Hazardous Substances ; metabolism ; poisoning ; Humans ; Mercury ; metabolism ; Metallothionein ; biosynthesis ; genetics ; metabolism ; Zinc ; metabolism

Heavy metal wastewater poses a serious threat to the environment. In comparison to the existing methods of chemical precipitation, ion exchange and carbon adsorption, biosorption is an attractive alternative for the recovery of heavy metals from industrial effluents. However, nickel ion, different from other heavy metal ions, is a more recalcitrant pollutant and has low affinity to many metal tolerant microorganisms. In this study, Escherichia coli JM109 was genetically engineered to simultaneously express a Ni2+ transport system (the product of nixA gene) andoverexpress metallothionein (MT). NixA protein has a high affinity for Ni2+, and metallothioneins (MTs) are capable of binding a variety of heavy metals including Ni2+ . The Ni2+ bioaccumulation performance of the genetically engineered E. coli JM109 was evaluated. Time-course test showed that the bioaccumulation rate was rapid, and 95% of the accumulation was achieved within the first 10 minutes. The maximum Ni2+ bioaccumulation by genetically engineered E. coli cells was dramatically increased from 1.54 mg/g to 10.11mg/g, a more than five-fold increase than that of the original E. coli strain. The isotherm was of Langmuir type. Within the tested pH range (pH 4-10), the engineered cells displayed more resistance to pH variation, retaining up to 80% of the Ni2+ binding capacity at pH 4, while the original E. coli host cells lost 80% of Ni2+ binding capacity at pH 4. The presence of Na+ and Ca2+ affected Ni2+ bioaccumulation, but the effects were not serious, as 71% and 66% of the Ni2+ binding capacities were retained respectively at the concentrations of 1000 mg/L Na+ and 1000 mg/L Ca2+ . However, Mg2+ exerted a severe adverse effect on Ni2+ bioaccumulation, 83% of Ni2+ accumulating capacity was lost when Mg2+ concentration reached 200 mg/L. The effects of different kinds of heavy metals on Ni2+ accumulating were different. The genetically engineered E. coli cell lost less than 45% of its Ni2+ bioaccumulation activity in the presence of 50 mg/L lead or cadmium, 66% in the presence of 25mg/L mercury and 84% in the presence of 40 mg/L copper. The presence of glucose did not improve Ni2+ uptake. Our study suggests that the genetically engineered E. coli JM109 has potential application for effective and efficient recovery of nickel from aqueous solutions.
Biodegradation, Environmental ; Cation Transport Proteins ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Engineering ; methods ; Hydrogen-Ion Concentration ; Metallothionein ; genetics ; metabolism ; Nickel ; metabolism ; Water Pollutants, Chemical ; metabolism