Objective: To study the effects of cadmium chloride (CdCl(2)) exposure on testicular autophagy levels and blood-testis barrier integrity in prepubertal male SD rats and testicular sertoli (TM4) cells. Methods: In July 2021, 9 4-week-old male SD rats were randomly divided into 3 groups: control group (normal saline), low dose group (1 mg/kg·bw CdCl(2)) and high dose group (2 mg/kg·bw CdCl(2)), and were exposed with CdCl(2) by intrabitoneal injection. 24 h later, HE staining was used to observe the morphological changes of testis of rats, biological tracer was used to observe the integrity of blood-testis barrier, and the expression levels of microtubule-associated protein light chain 3 (LC3) -Ⅰ and LC3-Ⅱ in testicular tissue were detected. TM4 cells were treated with 0, 2.5, 5.0 and 10.0 μmol/L CdCl(2) for 24 h to detect the toxic effect of cadmium. The cells were divided into blank group (no exposure), exposure group (10.0 μmol/L CdCl(2)), experimental group[10.0 μmol/L CdCl(2)+60.0 μmol/L 3-methyladenine (3-MA) ] and inhibitor group (60.0 μmol/L 3-MA). After 24 h of treatment, Western blot analysis was used to detect the expression levels of LC3-Ⅱ, ubiquitin binding protein p62, tight junction protein ZO-1 and adhesion junction protein N-cadherin. Results: The morphology and structure of testicular tissue in the high dose group were obvious changed, including uneven distribution of seminiferous tubules, irregular shape, thinning of seminiferous epithelium, loose structure, disordered arrangement of cells, abnormal deep staining of nuclei and vacuoles of Sertoli cells. The results of biological tracer method showed that the integrity of blood-testis barrier was damaged in the low and high dose group. Western blot results showed that compared with control group, the expression levels of LC3-Ⅱ in testicular tissue of rats in low and high dose groups were increased, the differences were statistically significant (P<0.05). Compared with the 0 μmol/L, after exposure to 5.0, 10.0 μmol/L CdCl(2), the expression levels of ZO-1 and N-cadherin in TM4 cells were significantly decreased, and the expression level of p62 and LC3-Ⅱ/LC3-Ⅰ were significantly increased, the differences were statistically significant (P<0.05). Compared with the exposure group, the relative expression level of p62 and LC3-Ⅱ/LC3-Ⅰ in TM4 cells of the experimental group were significantly decreased, while the relative expression levels of ZO-1 and N-cadherin were significantly increased, the differences were statistically significant (P<0.05) . Conclusion: The mechanism of the toxic effect of cadmium on the reproductive system of male SD rats may be related to the effect of the autophagy level of testicular tissue and the destruction of the blood-testis barrier integrity.
Rats ; Male ; Animals ; Testis ; Cadmium Chloride/metabolism* ; Cadmium ; Blood-Testis Barrier/metabolism* ; Rats, Sprague-Dawley ; Cadherins/metabolism* ; Autophagy

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

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 ; Adsorption ; Cadmium ; toxicity ; Calcium Chloride ; pharmacology ; Peroxidase ; metabolism ; Plant Roots ; drug effects ; enzymology ; growth & development ; Stress, Physiological ; drug effects ; Superoxide Dismutase ; metabolism ; Wedelia ; drug effects ; enzymology ; growth & development ; metabolism

This study investigated the conjoined cellular oxidative damage of human embryo kidney 293T (HEK293T) cells induced by cadmium chloride (CdCl(2)) and nanometer titanium dioxide (nano-TiO(2)). RT-PCR technique was used to detect the expressions of Heme oxygenase-1 (HO-1) and 8-oxoguanine DNA glycosylase (OGG1). The activities of superoxide dismutase (SOD) and catalase enzyme (CAT) and concentrations of reactive oxygen species (ROS) and maldondialdehyde (MDA) were measured by different approaches. The results showed that CdCl(2) and nano-TiO(2) at a low concentration of 0.75 total toxic unit (TU) exerted an additive effects on HO-1 gene expression, CAT activities and MDA concentrations. When the total TU was increased to 1 or 1.25 TU, the interaction was synergetic. Moreover, the mixture with high proportion of CdCl(2) produced an additive effect on the OGG1 gene expression, and the interaction was changed to be synergetic when the concentration of CdCl(2) was lower than or equal to that of nano-TiO(2). Synergetic effects of CdCl(2) and nano-TiO(2) on cellular oxidative damage of HEK293T cells were found as indicated by the changes in the SOD activities and ROS concentrations. It was concluded that CdCl(2) and nano-TiO(2) exerts synergistic effects on the cellular oxidative damage of HEK293T cells, and the sensitivity of these indicators of oxidative damage varies with the proportion of CdCl(2) and nano-TiO(2) in the mixture.
Cadmium Chloride ; toxicity ; Drug Synergism ; HEK293 Cells ; Humans ; Kidney ; cytology ; Nanoparticles ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Titanium ; toxicity

OBJECTIVETo examine the presence of gender differences in pulmonary inflammation evoked by acute systemic cadmium administration in rats.

METHODSPresence of basic indicators of lung inflammation (inflammatory cytokine lung content, leukocyte infiltration and activity of cells recovered from lungs by enzyme digestion) was analyzed and compared in animals of the two sexes.

RESULTSIntraperitoneal administration of cadmium (1.0 mg/kg) resulted in higher cadmium content in lungs of female rats. Higher tumor necrosis factor (TNF) content was noted in lung homogenates of male rats, while interleukin-6 (IL-6) content was slightly, but significantly greater in lungs of female rats. Increased leukocyte infiltration was observed in lungs of male rats, mainly due to neutrophils. Increased responsiveness to phorbol myristate acetate (PMA) stimulation was noted in cells recovered from lungs of male rats. Rise in intracellular content of myeloperoxidase (MPO) was noted in lung cells from cadmium-treated rats of both sexes, but higher in cells from male rats.

CONCLUSIONSPresented data documented a more intense pulmonary inflammatory response to systemic cadmium administration in males, with higher IL-6 levels in lungs of female individuals. These sex differences in proinflamatory activity of cadmium in lungs should be taken into consideration in studying the remote toxicity of this heavy metal.

Animals ; Cadmium Chloride ; pharmacokinetics ; toxicity ; Cytokines ; immunology ; Environmental Pollutants ; pharmacokinetics ; toxicity ; Enzyme-Linked Immunosorbent Assay ; Female ; Leukocyte Count ; Leukocytes ; cytology ; immunology ; Lung ; drug effects ; immunology ; metabolism ; Male ; Neutrophil Infiltration ; immunology ; Peroxidase ; metabolism ; Pneumonia ; chemically induced ; immunology ; metabolism ; Rats ; Rats, Inbred Strains ; Sex Characteristics

To study if Solanum nigrum hairy roots can be used for phytoremediation of Cd contamination, we investigated the effects of cadmium (Cd) alone, and in combination with different concentrations of CaCl2, on growth, activities of superoxide dismutase (SOD) and peroxidase (POD) and Cd absorption by hairy roots of S. nigrum L. var pauciflorum. The results showed that Cd concentrations of lower than 50 micromol/L enhanced the growth of hairy roots, while higher than 100 micromol/L inhibited growth and decreased the number of branched roots, also causing the root tips to become brown and shorter in length. In comparison with a control, the soluble protein content, the activities of SOD and POD in hairy roots cultures showed a trend of first increased and then gradually decreased, while the malondialdehyde (MDA) content significantly increased, when increasing the Cd concentrations. Cd concentration of 100 micromol/L or 300 micromol/L in combination with 10-30 mmol/L CaCl2 resulted in a decreased content of soluble protein and MDA in the hairy roots, but an enhanced SOD activity. The increased POD activities were observed when cultured in 100 micromol/L Cd and 10-30 mmol/L CaCl2 but decreased when cultured in 300 micromol/L Cd and 10-30 mmol/L CaCl2. Atomic Absorption Spectrometry determination showed that the Cd absorbed and adsorbed by the hairy roots increased along with the increase of Cd concentration. The exogenous addition of 10-30 mmol/L CaCl2 could reduce the toxicity of Cd. This was achieved on one hand by reducing the absorption of Cd, on the other hand by decreasing the lipid peroxidation through regulating the activities of antioxidant enzymes SOD and POD in the hairy roots.
Absorption ; Biodegradation, Environmental ; Cadmium ; isolation & purification ; metabolism ; Calcium Chloride ; metabolism ; Peroxidase ; metabolism ; Plant Roots ; growth & development ; physiology ; Soil Pollutants ; isolation & purification ; metabolism ; Solanum nigrum ; enzymology ; growth & development ; physiology ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the alternative expression and sequence of human elongation factor-1 delta (human EF-1 delta p31) during malignant transformation of human bronchial epithelial cells induced by cadmium chloride (CdC12) and its possible mechanism.

METHODSTotal RNA was isolated at different stages of transformed human bronchial epithelial cells (16HBE) induced by CdCl2 at a concentration of 5.0 microM. Special primers and probe for human EF-1 delta p31 were designed and expression of human EF-1 delta mRNA from different cell lines was detected with fluorescent quantitative PCR technique. EF-18 cDNA from different cell lines was purified and cloned into pMD 18-T vector followed by confirming and sequencing analysis.

RESULTSThe expressions of human EF-1 beta p31 at different stages of 16HBE cells transformed by CdCl2 was elevated (P < 0.01 or P < 0.05). Compared with their corresponding non-transformed cells, the overexpression level of EF-1 delta p31 was averagely increased 2.9 folds in Cd-pretransformed cells, 4.3 folds in Cd-transformed cells and 7.2 folds in Cd-tumorigenic cells. No change was found n the sequence of overexpressed EF-1beta p31 at different stages of 16HBE cells transformed by CdCl2.

CONCLUSIONOverexpression of human EF-1beta p31 is positively correlated with malignant transformation of 16HBE cells induced by CdC12, but is not correlated with DNA mutations.

Cadmium Chloride ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; metabolism ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Peptide Elongation Factor 1 ; genetics ; metabolism ; Respiratory Mucosa ; drug effects ; metabolism ; pathology ; Sequence Analysis, DNA

OBJECTIVETo explore the effect of cadmium chloride on the expression and phosphorylation of mitogen-activated protein kinase (MAPK) in normal rat kidney epithelial (NRK) cells.

METHODSThe NRK cells were incubated with cadmium chloride either at respective dose (0, 1, 5, 10, 20, 40 µmol/L) for 24 h or at same dose (10 µmol/L) for respective time (0, 0.5, 1.0, 2.0, 4.0, 8.0 h). Western blotting was applied to test the expression of MAPK in NRK cells (ERK1/2, p38, JNK); and phosphor-specific antibody to detect the phosphorylated MAPK (p-ERK1/2, p-p38, p-JNK).

RESULTSThere was no significant difference in the MAPK expression among the groups either treated with different doses or for different time; however, the level of phosphorylated MAPK was comparatively higher than it in control group. There was an obvious expression of p-ERK1/2 at 1.00 ± 0.06 in the group incubated with 10 µmol/L CdCl(2); and the expression in the 20 µmol/L and 40 µmol/L CdCl(2) group was 2.58 ± 0.11, 2.76 ± 0.23 respectively, which was 1.58 and 1.76 times more than it in 10 µmol/L CdCl(2) group. The differences were statistically significant (F = 827.70, P < 0.01). The respective expression of p-p38MAPK in the 20 µmol/L (2.47 ± 0.20)and 40 µmol/L CdCl(2) group (3.73 ± 0.25)was 1.47 and 2.73 times more than it in control group (1.00 ± 0.02), and the differences were also statistically significant (F = 280.06, P < 0.01). There was a dose-effect relationship of the concentration of cadmium in the expression of p-ERK1/2 (r = 0.919, t = 4.69, P = 0.009) and p-p38MAPK (r = 0.945, t = 5.79, P = 0.004). Additionally, phosphorylated MAPK expressed in a time-dependent manner. The expression of p-ERK1/2 was obvious in the group incubated for 1 h (1.26 ± 0.11), and the respective expression in the 4 h group (1.51 ± 0.07) and 8 h group (3.53 ± 0.23) was 1.5 and 3.5 times of it in the control group (1.00 ± 0.02). The differences were statistically significant (F = 427.82, P < 0.001). The expression of p-p38MAPK increased significantly in 1 h group (1.31 ± 0.07); while the respective expression in 4 h group (3.53 ± 0.32) and 8 h group (4.41 ± 0.38) was 3.5 and 4.4 times of it in control group (1.00 ± 0.03). The differences were also statistically significant (F = 280.06, P < 0.001).

CONCLUSIONCadmium chloride could significantly enhance the phosphorylation of MAPK in NRK cells; and it is probably associated with the activation of MAPK.

Animals ; Cadmium Chloride ; toxicity ; Cell Line ; Epithelial Cells ; drug effects ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; Phosphorylation ; Rats ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo explore the expression and sequence of ERCC1 gene in CdCl2-induced transformed human bronchial epithelial 16HBE cells at different stages.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemcial staining (SP method) were used to measure the ERCC1 mRNA and protein expression in 16HBE cells at different passages treated with CdCl2 (the 5th, 15th, 35th passage, and neoplastic cells from tumors formed in nude mice). ERCC1 exon 3,exon 4 of the 16HBE cells and tumor cells from nude mice were amplified by polymerase chain reactions (PCR), the amplified DNA strips were purified,and the exons were detected by DNA analysis.

RESULTSDuring the passages of 16HBE cells treated with CdCl2, the expression of ERCC1 gene was decreased gradually. The ERCC1 gene mRNA and protein expression levels of the CdCl2-transformed 35th passage 16HBE cells and tumor cells from nude mice were significantly decreased comparing with those in non-transformed 16HBE cells (P < 0.01). In the CdCl2-induced tumorigenic cells in nude mice, there was adenine (A) deletion in 1st site of ERCC1 exon 4. The mutation was frame shift mutation.

CONCLUSIONThe decreased expression and mutation of ERCC1 gene may be the possible carcinogenic mechanism of CdCl2.

Animals ; Bronchi ; cytology ; Cadmium Chloride ; toxicity ; Cell Transformation, Neoplastic ; Cells, Cultured ; DNA-Binding Proteins ; genetics ; metabolism ; Endonucleases ; genetics ; metabolism ; Epithelial Cells ; cytology ; metabolism ; Exons ; Frameshift Mutation ; Humans ; Mice ; Mice, Nude ; RNA, Messenger ; metabolism

OBJECTIVETo study apoptosis induced by cadmium chloride (CdCl2) and the alteration in activity of protein kinase B (PKB/Akt) in adrenocortical cells.

METHODSFasciculata-glomerulosa (FG) cells of male guinea pigs were dispersed and primarily cultured in vitro. Features of apoptotic cells were observed using combined labeling with annexin-V and propidium iodide (PI) and flow cytometry. The activity of PKB/Akt was determined with immunoprecipitation and the chemiluminescence assay.

RESULTSApoptosis rate of FG cells increased with dose of CdCl2 two hours after treatment with 6.25-100.00 micromol/L of it, with significant difference in the groups treated with 25.00, 50.00, 100.00, micromol/L of CdCl2, as compared with the control group (P < 0.01). Regression analysis showed that occurrence of apoptosis correlated with the dose of CdCl2 in a dose-response pattern. In the meanwhile, there were obviously elevated percentages of apoptotic cells as the increase in duration of incubation ranging from 5.58% to 73.08% for incubating cells with 50.00 micromol/L of CdCl2, from 15 minutes to 4 hours. Duration of incubating cells with CdCl2 were correlated with occurrence of apoptosis in a time-effect manner. The gray scales of PKB/Akt were manifested to be decreased as the ascending of CdCl2 dosage from 6.25 to 200.00 micromol/L, with the linear correlation.

CONCLUSION6.25 to 100.00 micromol/L CdCl2 might elicit apoptosis of adrenocortical cells. Meanwhile, PKB/Akt is decreased.

Adrenal Cortex ; drug effects ; enzymology ; pathology ; Animals ; Apoptosis ; drug effects ; Cadmium Chloride ; toxicity ; Cells, Cultured ; Guinea Pigs ; Male ; Proto-Oncogene Proteins c-akt ; metabolism