1.Effect of ultrafiltration-membrane extracts of Radix Rehmanniae Praeparata on proliferation and genetic stability of bone marrow-derived mesenchymal stem cells induced by cadmium chloride.
Yong-qi LIU ; Qi ZHANG ; Li JING-YA ; Rui DA ; Ya-li LUO ; Yun SU ; Zhi-wei WU ; Chun-lul YAN ; Lei NIE
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(4):450-456
OBJECTIVETo study the effect of ultrafiltration-membrane extracts of Radix Rehmanniae Praeparata (UMERRP) on theproliferation and genetic stability of bone marrow-derived mesenchymal stem cells (BMSCs) induced by cadmium chloride (CdCl2).
METHODSProtective effects on the proliferation, micronuclear rates, chromosome aberration rates, and apoptosis rates were observed by micronuclei test, karyotype analysis, and flow cytometry.
RESULTSCompared with the CdCl2 group, UMERRP with different molecular weights at 0. 8 g/L could obviously promote the proliferation (P <0. 05). Compared with the control group, micronuclear rates, chromosome aberration rates, and apoptosis rates were obviously enhanced in the CdCl2 group (P <0. 05). Compared with the CdCl2 group, UMERRP with different molecular weights could obviously decreased CdCl2 induced micronuclear rates, chromosome aberration rates, and apoptosis rates (P <0. 05). Of them, BMSC micronuclear rates and chromosome aberration rates decreased most obvious in UMERRP groups with molecular weight below 10 000 (P <0. 05). The apoptosis rate decreased most obviously in UMERRP groups with molecular weight ranging 100 000 and 200 000 (P <0. 05).
CONCLUSIONUMERRP could reduce CdCl2 induced micronuclear rates, chromosome aberration rates, and apoptosis rates.
Apoptosis ; Bone Marrow ; Cadmium Chloride ; toxicity ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Hematopoietic Stem Cells ; Humans ; Mesenchymal Stromal Cells ; Ultrafiltration
2.Protective effect of sesamin against myocardial injury induced by cadmium chloride in rats.
Junzhi TIAN ; Rong ZHANG ; Hongxue ZHANG ; Yi LIU ; Yujie NIU ; Lijuan ZHAO ; Luqi WANG ; Huiccai GUO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2014;32(5):357-360
OBJECTIVETo investigate the protective effect of sesamin against cadmium chloride (CdCl2)-induced cardiotoxicity in rats.
METHODSFifty male Wistar rats were randomly assigned to five groups: control group, CdCl2 group, and low-, middle-, and high-dose sesamin groups. The control group was given normal saline. The CdCl2 group and sesamin groups were intraperitoneally injected with CdCl2 (5 mg/kg×2 d), and the low-, middle-, and high-dose sesamin groups were given 20, 40, and 80 mg/kg sesamin, respectively. All treatments lasted for four weeks. ECG was measured by a physiological recorder, and serum myocardial enzyme levels were determined by biochemical assay. The heart was weighed, and heart tissues were used in histopathological examination and determination of malondialdehyde (MDA) level.
RESULTSCompared with the control group, the CdCl2 group showed significantly higher levels of serum CK and CK-MB, an increased heart coefficient, significant ST-segment elevation, and higher level of MDA in myocardial tissue (P < 0.05). Histopathological analysis showed edema of myocardial tissues and cells, myocardial fibers disorder, karyopyknosis, and uneven or deep staining of nuclear chromatin. Different doses of sesamin relieved the myocardial pathological changes induced by CdCl2, and high-dose sesamin was the most effective. The middle- and high-dose sesamin groups showed significantly reduced serum CK and CK-MB levels compared with the CdCl2 group (P < 0.05). The heart coefficient of the high-dose sesamin group (0.19±0.01%) was significantly lower than that of the CdCl2 group (0.21±0.01%) (P < 0.05). Myocardial MDA levels of the three sesamin groups (42.32±4.65, 36.71±5.34, and 33.12±4.62 nmol/mg pro, respectively) were all significantly lower than that of the CdCl2 group (55.87±3.65 nmol/mg pro) (P < 0.05).
CONCLUSIONSesamin can relieve myocardial injury induced by CdCl2, and one possible mechanism is the enhancement of antioxidant capacity of myocardial tissue.
Animals ; Cadmium Chloride ; toxicity ; Creatine Kinase, MB Form ; blood ; Dioxoles ; pharmacology ; Heart ; drug effects ; Lignans ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Wistar
3.Protection of cadmium chloride induced DNA damage by Lamiaceae plants.
Ramaraj THIRUGNANASAMPANDAN ; Rajarajeswaran JAYAKUMAR
Asian Pacific Journal of Tropical Biomedicine 2011;1(5):391-394
OBJECTIVETo analyze the total phenolic content, DNA protecting and radical scavenging activity of ethanolic leaf extracts of three Lamiaceae plants, i.e. Anisomelos malabarica (A. malabarica), Leucas aspera (L. aspera) and Ocimum basilicum (O. basilicum).
METHODSThe total polyphenols and flavonoids were analyzed in the ethanolic leaf extracts of the lamiaceae plants. To determine the DNA protecting activity, various concentrations of the plant extracts were prepared and treated on cultured HepG2 human lung cancer cells. The pretreated cells were exposed to H2O2 to induce DNA damage through oxidative stress. Comet assay was done and the tail length of individual comets was measured. Nitric oxide and superoxide anion scavenging activities of lamiaceae plants were analyzed.
RESULTSAmong the three plant extracts, the highest amount of total phenolic content was found in O. basilicum (189.33 mg/g), whereas A. malabarica showed high levels of flavonoids (10.66 mg/g). O. basilicum also showed high levels of DNA protecting (85%) and radical scavenging activity.
CONCLUSIONSThe results of this study shows that bioactive phenols present in lamiaceae plants may prevent carcinogenesis through scavenging free radicals and inhibiting DNA damage.
Cadmium Chloride ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Free Radical Scavengers ; chemistry ; pharmacology ; Hep G2 Cells ; Humans ; Lamiaceae ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; Plant Leaves ; chemistry ; Protective Agents ; chemistry ; pharmacology
4.Effects of selenium on rat hepatocellular DNA damage, apoptosis and changes of cell cycle induced by cadmium in vivo.
Chinese Journal of Preventive Medicine 2004;38(3):155-158
OBJECTIVEThis study was conducted to study the effects of sodium selenite on rat hepatocellular DNA damage, apoptosis, changes of cell cycle and DNA relative content induced by cadmium chloride in vivo.
METHODSBoth sodium selenite at the dose of 5 micromol/kg and cadmium chloride at the dose of 5 micromol/kg, 10 micromol/kg and 20 micromol/kg were given to rats by i.p. and there were 5 male SD rats in each group. Hepatocellular DNA damage was measured by the single cell gel electrophoresis (or comet assay), hepatocellular apoptosis was measured with TUNEL (TdT-mediated dUTP Nick End Labelling) and flow cytometry, DNA relative content (DNA(RC)) and cell cycle were detected with flow cytometry.
RESULTSWhen sodium selenite at the dose of 5 micromol/kg acted jointly with cadmium chloride at the dose of 5 micromol/kg, 10 micromol/kg and 20 micromol/kg respectively, the results showed that selenium reduced the effect of cadmium on DNA damage and apoptosis and decreased the rates of DNA damage and the rates of apoptosis significantly. Sodium selenite at the dose of 5 micromol/kg increased cell number of G(0)/G(1) period decreased by cadmium chloride at the dose of 5 micromol/kg and increased cell number of G(2)/M period decreased by cadmium chloride at the dose of 10 micromol/kg and 20 micromol/kg. Sodium selenite at the dose of 5 micromol/kg increased DNA relative content reduced by cadmium chloride at the dose of 10 micromol/kg and 20 micromol/kg.
CONCLUSIONSIt was suggested that selenium at certain doses could antagonize DNA damage, apoptosis, changes of cell cycle and DNA relative content induced by cadmium in rat hepatocytes in vivo.
Animals ; Apoptosis ; drug effects ; Cadmium Chloride ; antagonists & inhibitors ; toxicity ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; DNA Damage ; drug effects ; Dose-Response Relationship, Drug ; Hepatocytes ; drug effects ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Sodium Selenite ; pharmacology
5.Malignant transformation and abnormal expression of eukaryotic initiation factor in bronchial epithelial cells induced by cadmium chloride.
Yi-Xiong LEI ; Lian WEI ; Min WANG ; Gen-Rong WU ; Min LI
Biomedical and Environmental Sciences 2008;21(4):332-338
OBJECTIVETo analyze the relationship between malignant transformation and abnormal expression of eukaryotic initiation factor 3 (eIF3 p36) in human bronchial epithelial (16HBE) cells induced by cadmium chloride (CdCl2).
METHODS16HBE cells were treated several times with different concentrations of CdCl2. Tumorigenic potential of transformed cells was identified by assays for anchorage-independent growth in soft agar and for tumorigenicity in nude mice after the 35th passage. Total RNA was isolated from 16HBE cells induced by CdCl2, including non-transformed, Cd-transformed, and Cd-tumorigenic cell lines. Special primers for eIF3 p36 were designed and the expression of eIF3 mRNA in different cell lines was detected with fluorescent quantitative-polymerase chain reaction technique (FQ-PCR).
RESULTSThe 35th passage of 16HBE cells transformed by CdCl2 exhibited overlapping growth. Compared with the non-transformed cells, colonies of transformed cell lines in soft agar showed statistically significant increases and dose-dependent effects (P<0.01). All Cd-induced transformed cell lines formed tumors in nude mice within 2 weeks of inoculation, but none of the mice injected with non-transformed cells showed tumors even after 3 weeks. All tumors were pathologically identified as poorly differentiated squamous cell carcinoma. The eIF3 p36 genes in different stages of 16HBE cells transformed by CdCl2 were elevated as compared with the non-transformed control (P<0.01), and the eIF3 expression increased with the degree of cell malignancy.
CONCLUSIONCdCl2 is capable of inducing morphological transformation in 16HBE cells and transformed cells are potentially tumorigenic. Over-expression of eIF3 p36 is positively correlated with malignant transformation of 16HBE cells induced by CdCl2 and may be one of the molecular mechanisms potentially responsible for carcinogenesis due to Cd.
Animals ; Base Sequence ; Bronchi ; cytology ; drug effects ; metabolism ; Cadmium Chloride ; pharmacology ; Cell Transformation, Neoplastic ; DNA Primers ; Epithelial Cells ; drug effects ; metabolism ; Eukaryotic Initiation Factors ; metabolism ; Humans ; Mice ; Mice, Nude ; Polymerase Chain Reaction
6.Alleviated affect of exogenous CaCl2 on the growth, antioxidative enzyme activities and cadmium absorption efficiency of Wedelia trilobata hairy roots under cadmium stress.
Heping SHI ; Yunling WANG ; PoKeung Eric TSANG ; LeeWah Andrew CHAN
Chinese Journal of Biotechnology 2012;28(6):747-762
In order to study the physiological mechanism of exogenous calcium on the toxicity of heavy metal cadmium (Cd) to Wedelia trilobata hairy roots, the effects of Cd alone, and in combination with different concentrations of Ca on growth, contents of soluble protein and malondialdehyde (MDA), activities of superoxide dismutase (SOD) and peroxidase (POD), Cd2+ absorption in W. trilobata hairy roots were investigated. Cd concentrations lower than 50 micromol/L enhanced the growth of hairy roots, while concentrations higher than 100 micromol/L inhibited growth, making the branched roots short and small, and also turning the root tips brown, even black. In comparison with the control (0 micromol/L Cd), the soluble protein content in hairy roots was found to increase when cultured with 10-50 micromol/L Cd, and decrease when exposed to a cadmium concentration higher than 100 micromol/L Cd. In addition, the activities of POD and SOD activity and MDA content were significantly higher than the control. Compared to the control (hairy roots cultured without 10-30 mmol/L Ca), 100 micromol/L Cd or 300 micromol/L Cd in combination with 10-30 mmol/L Ca resulted in increased growth, causing the main root and secondary roots thicker and also an increase in soluble protein content. On the contrary, MDA content and POD and SOD activities decreased. Quantitative analysis by Atomic Absorption Spectrophotometry showed that W. trilobata hairy roots can absorb and adsorb heavy metal Cd in the ionic form of Cd2+. The maximum content of Cd2+ absorbed by the hairy roots was obtained with a concentration 100 micromol/L Cd2+ while that of Cd2+ adsorbed by hairy roots was achieved with a concentration of 300 micromol/L Cd2+. The exogenous addition of 10-30 mmol/L Ca2+ was found to reduce the absorption, adsorption of Cd2+ and the toxicity of Cd significantly. This reduction in toxicity was caused by the reduction in the absorption of Cd and decreasing the lipid peroxidation through regulating the activities of antioxidant enzymes SOD and POD in the hairy roots.
Absorption
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Adsorption
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Cadmium
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toxicity
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Calcium Chloride
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pharmacology
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Peroxidase
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metabolism
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Plant Roots
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drug effects
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enzymology
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growth & development
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Stress, Physiological
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drug effects
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Superoxide Dismutase
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metabolism
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Wedelia
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drug effects
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enzymology
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growth & development
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metabolism
7.Chelation in metal intoxication XLVI: synthesis of some alpha-mercapto-beta-substituted aryl acrylic acids and their in vitro cadmium chelating ability.
Madhumita CHATTERJEE ; Vinod K DWIVEDI ; Kirti KHANDEKAR ; Sushil K TANDON
Biomedical and Environmental Sciences 2004;17(1):27-32
OBJECTIVETo synthesize some new alpha-mercapto-beta-substituted aryl acrylic acids, characterize them and investigate their in vitro cadmium chelating ability.
METHODSSix alpha-mercapto-beta-substituted aryl acrylic acids were prepared by the alkaline hydrolysis of 5- (aryl methylene) rhodanines, obtained from the condensation of substituted aldehydes and rhodanine following the reported procedure. The new compounds were characterized by elemental analysis, infrared (IR) and nuclear magnetic resonance (NMR) spectroscopy. The liver and kidney from cadmium chloride pre-administered rats were homogenized and their nuclear mitochondrial fraction (NMF) and supernatant cytosol fraction (SCF) were separated. A measured volume of each fraction was dialyzed separately using "dialysis sack" against buffered-KCl medium containing a compound in the final concentration of 1 x 10(-3) mol/L for 3 h at 37 degrees C. The whole content of "sack" was subjected to cadmium estimation following digestion with conc. Nitric acid was detected using flame atomic absorption spectrometer.
RESULTSThe in vitro screening showed that alpha-mercapto-beta-(p-methoxyphenyl) acrylic acid (compound 2) and alpha-mercapto-beta-(m-methoxy, p-hydroxyphenyl) acrylic acid (compound 4) were more effective than alpha-mercapto-beta-thienyl acrylic acid (compound 1) and alpha-mercapto-beta-(p-dimethylaminophenyl) acrylic acid (compound 3) in mobilizing cadmium as their dialyzable chelates. The presence of a methoxy group on the phenyl moiety (compounds 2 and 4) increases the metal chelating ability of mercapto acrylic acids.
CONCLUSIONSCompounds 2 and 4 seem to have accessibility to the cellular system and capability of chelating-out the intracellularly bound cadmium.
Acrylates ; chemical synthesis ; chemistry ; pharmacology ; Animals ; Cadmium Chloride ; metabolism ; toxicity ; Chelating Agents ; chemical synthesis ; chemistry ; pharmacology ; Injections, Intraperitoneal ; Kidney ; drug effects ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Mitochondria ; drug effects ; metabolism ; Mitochondria, Liver ; drug effects ; metabolism ; Rats ; Sulfhydryl Compounds ; chemical synthesis ; chemistry ; pharmacology
8.Evidence for Cyclooxygenase-2 Association with Caveolin-3 in Primary Cultured Rat Chondrocytes.
Jin Oh KWAK ; Woon Kyu LEE ; Hyun Woo KIM ; Sun Mi JUNG ; Kwang Jin OH ; Sang Yong JUNG ; Yang Hoon HUH ; Seok Ho CHA
Journal of Korean Medical Science 2006;21(1):100-106
The purpose of this study was to demonstrate the cellular localization of cyclooxygenase-2 (COX-2) and caveolin-3 (Cav-3) in primarily cultured rat chondrocytes. In normal rat chondrocytes, we observed relatively high levels of Cav-3 and a very low level of COX-2 mRNA and protein. Upon treating the chondrocytes with 5 microM of CdCl2 (Cd) for 6 hr, the expressions of COX-2 mRNA and protein were increased with the decreased Cav-3 mRNA and protein expressions. The detergent insoluble caveolae-rich membranous fractions that were isolated from the rat chondrocytes and treated with Cd contained the both proteins of both COX-2 and Cav-3 in a same fraction. The immuno-precipitation experiments showed complex formation between the COX-2 and Cav-3 in the rat chondrocytes. Purified COX-2 with glutathione S-transferase-fused COX-2 also showed complex formation with Cav-3. Confocal and electron microscopy also demonstrated the co-localization of COX-2 and Cav-3 in the plasma membrane. The results from our current study show that COX-2 and Cav-3 are co-localized in the caveolae of the plasma membrane, and they form a protein-protein complex. The co-localization of COX-2 with Cav-3 in the caveolae suggests that the caveolins might play an important role for regulating the function of COX-2.
Animals
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Animals, Newborn
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Blotting, Western
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Cadmium Chloride/pharmacology
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Caveolae/drug effects/metabolism/ultrastructure
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Caveolin 3/*genetics/metabolism
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Cell Membrane/drug effects/metabolism
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Cells, Cultured
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Chondrocytes/cytology/drug effects/*metabolism
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Cyclooxygenase 2/*genetics/metabolism
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Gene Expression
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Immunoprecipitation
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Microscopy, Confocal
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Microscopy, Electron
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RNA, Messenger/genetics/metabolism
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Rats
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Reverse Transcriptase Polymerase Chain Reaction