Selenium modulates AR/IGF-1R/EGFR and TROP2 signaling pathways and improves anticancer efficacy in murine mammary carcinoma 4T1.

The micronutrient selenium (Se) has been shown to exert potential anticancer properties. This study aimed to evaluate the effects of Se (in Se yeast form) on the selenoproteins (SELENO), AR/IGF-1R/EGFR, PI3K/Akt/mTOR and Ras/Raf/ERK cascades, and immune checkpoint blockade in TNBC murine 4T1 cells. We also assessed the effects of combination treatment with chemotherapeutic doxorubicin and Se on trophoblast cell surface antigen 2 (TROP2) levels. Compared with the control groups, cells incubated with Se (0.25, 0.5, 0.75, 1.0, 1.5 µg Se/mL) have lower viability, raised intracellular Se concentrations and SELENO expression, and higher malondialdehyde products in a dose-dependent manner. Se induced the inactivation of AR/IGF-1R/EGFR and downregulation of the PI3K/Akt/mTOR and Ras/Raf/ERK signaling molecules. Se-treated cells also exhibited decreased mitochondrial membrane potential, reduced levels of the cell cycle regulatory protein cyclin D1, cancer stemness, metastatic and EMT-related markers, and increased apoptosis. Subsequently, Se treatment significantly suppressed PD-1/PD-L1 and CTLA-4 mRNA levels and proteins. Doxorubicin decreased 4T1 cell viability and TROP2 expression levels, but the addition of Se to doxorubicin contributed to further reductions. Similar responses to Se treatment were also observed in the human MDA-MB-231 and MCF-7 breast cancer cells. These results show that Se upregulates SELENO and anti-AR/IGF-1R/EGFR signaling in TNBC cells, thus inducing oxidative stress-dependent apoptosis and cell cycle arrest, stemness, EMT, and metastasis, as well as blocking the immune checkpoint molecules. TROP2 down-regulation with Se is also a potential anti-TNBC therapeutic target.

Targeting EGFR in Combination with Nutritional Supplements on Antitumor Efficacy in a Lung Cancer Mouse Model.

Selenium (Se) and fish oil (FO) exert anti-epidermal growth factor receptor (EGFR) action on tumors. This study aimed to compare the anti-cancer efficacy of EGFR inhibitors (gefitinib and erlotinib) alone and in combination with nutritional supplements of Se/FO in treating lung cancer. Lewis LLC1 tumor-bearing mice were treated with a vehicle or Se/FO, gefitinib or gefitinib plus Se/FO, and erlotinib or erlotinib plus Se/FO. The tumors were assessed for mRNA and protein expressions of relevant signaling molecules. Untreated tumor-bearing mice had the lowest body weight and highest tumor weight and volume of all the mice. Mice receiving the combination treatment with Se/FO and gefitinib or erlotinib had a lower tumor volume and weight and fewer metastases than did those treated with gefitinib or erlotinib alone. The combination treatment exhibited greater alterations in receptor signaling molecules (lower EGFR/TGF-ß/TßR/AXL/Wnt3a/Wnt5a/FZD7/ß-catenin; higher GSK-3ß) and immune checkpoint molecules (lower PD-1/PD-L1/CD80/CTLA-4/IL-6; higher NKp46/CD16/CD28/IL-2). These mouse tumors also had lower angiogenesis, cancer stemness, epithelial to mesenchymal transitions, metastases, and proliferation of Ki-67, as well as higher cell cycle arrest and apoptosis. These preliminary results showed the Se/FO treatment enhanced the therapeutic efficacies of gefitinib and erlotinib via modulating multiple signaling pathways in an LLC1-bearing mouse model.

Fish Oil and Selenium with Doxorubicin Modulates Expression of Fatty Acid Receptors and Selenoproteins, and Targets Multiple Anti-Cancer Signaling in Triple-negative Breast Cancer Tumors.

Omega-3 fatty acids from fish oil (FO) and selenium (Se) potentiate some conventional therapies and have anticancer immune potential. This study aims to determine whether FO/Se modulates G-protein-coupled polyunsaturated fatty acid receptors (GPR-40 and GPR-120) and selenoproteins (Sel-H, Sel-W, and GPx4), and increases the therapeutic effect of doxorubicin in a dose-dependent manner on triple-negative breast cancer (TNBC) mouse. Mice were randomized into 5 groups (n = 7/group) and treated with physiological saline (control), low-dose doxorubicin, and doxorubicin in combination with low, medium, or high doses of FO/Se. The expression of signaling molecules in tumors was determined by measuring either mRNA or protein expression. Compared with doxorubicin alone, combination treatment resulted in lower tumor sizes and fewer overall metastasis, lower GPR-40 mRNA levels, and higher expression of all selenoproteins. Doxorubicin-FO/Se combination treatment decreased expression of membrane EGFR and FGFR, down-regulated downstream PI3K/AKT/mTOR, MAPK/ERK, and JAK2/c-Src/STAT3 signaling, increased tumor suppressor PTEN/TSC1/TSC2 expression and P53 activation, and suppressed oncogenic transcription factor expression. Dose-dependent inhibition of proliferation index Ki-67, cell cycle, and stem-cell-related markers were observed. Decreased immune check-points PD-L1/CTLA-4/Foxp3/CD86 and increased PD-1/CD28/IL-2 expression was also found. These observations suggest that the nutritional supplements FO/Se increase the chemotherapeutic efficacy of doxorubicin against TNBC by modulating GPR-40 and selenoprotein and targeting multiple signaling pathways in tumor tissues.

Combination of Fish Oil and Selenium Enhances Anticancer Efficacy and Targets Multiple Signaling Pathways in Anti-VEGF Agent Treated-TNBC Tumor-Bearing Mice.

Fish oil (FO) and selenium (Se) possess antiangiogenic potential in malignant tumors. This study aimed to determine whether combination of FO and Se enhanced treatment efficacy of low-dose antiangiogenic agent Avastin (bevacizumab) in a dose-dependent manner and targeted multiple signaling pathways in triple-negative breast cancer (TNBC)-bearing mice. Randomized into five groups, mice received treatment with either physiological saline (control), Avastin alone, or Avastin in combination with low, medium, and high doses of FO/Se. The target signaling molecules for anticancer were determined either by measuring protein or mRNA expression. Avastin-treated mice receiving FO/Se showed lower tumor growth and metastasis than did mice treated with Avastin alone. Combination-treated mice exhibited lower expressions in multiple proangiogenic (growth) factors and their membrane receptors, and altered cytoplasmic signaling molecules (PI3K-PTEN-AKT-TSC-mTOR-p70S6K-4EBP1, Ras-Raf-MEK-ERK, c-Src-JAK2-STAT3-TMEPAI-Smad, LKB1-AMPK, and GSK3ß/ß-catenin). Dose-dependent inhibition of down-stream targets including epithelial-to-mesenchymal transition transcription factors, nuclear cyclin and cyclin-dependent kinases, cancer stem cell markers, heat shock protein (HSP-90), hypoxia-inducible factors (HIF-1α/-2α), matrix metalloprotease (MMP-9), and increased apoptosis were observed. These results suggest that combination treatment with FO and Se increases the therapeutic efficacy of Avastin against TNBC in a dose-dependent manner through multiple signaling pathways in membrane, cytoplasmic, and nucleic targets.

Nutritional supplements in combination with chemotherapy or targeted therapy reduces tumor progression in mice bearing triple-negative breast cancer.

The potential anti-cancer properties of selenium (Se) and eicosapentaenoic acid (EPA)/docosahexaenoic acid (DHA) have been documented. However, few studies have been conducted examining anti-tumor effects of nutritional supplements (NS) containing Se and EPA/DHA in combination with anti-cancer agents, such as taxol (Tax), adriamycin (Adr), and avastin (Ava). Compared with triple-negative breast cancer (TNBC)-bearing positive control (TB) mice, a low dose of Tax, Adr, and Ava decreased tumor size and the incidence of metastasis in TB-Tax, TB-Adr, and TB-Ava groups. Combination treatment with anti-cancer agent and NS (2.7 µg Se and 5.1 mg EPA/3.7 mg DHA/g) induced additional decreases in TB-Tax-NS, TB-Adr-NS, and TB-Ava-NS groups. Th1-associated cytokines were increased, and Th2-type cytokines were decreased significantly in TB mice with combination treatment than that of anti-cancer agent treatment alone. Combination treatment with anti-cancer agents and NS has also been shown to further increased tumor malondialdehyde (MDA) levels, lowered hypoxia-inducible factor (HIF)-1α, angiogenic markers (vascular endothelial growth factor [VEGF] and CD31) and metastatic potential, as well as reduced heat shock proteins, receptor tyrosine kinase AXL, and surface markers of cancer stem cells, and increased apoptotic proteins. For immune checkpoint molecules, combination treatment was associated with a greater decrease in programmed cell death ligand-1 (PD-L1) in both tumors and mammary glands, but PD-1 level in primary tumors was increased. Our results suggest that combination treatment with low-dose anti-cancer agents (Tax, Adr, and Ava) and oral supplementation of Se/ EPA/DHA significantly decreased tumor growth and metastatic progression in TNBC mice through multiple anti-tumor mechanisms.

Altered Mineral Metabolism and Disequilibrium Between Calcification Promoters and Inhibitors in Chronic Hemodialysis Patients.

Patients undergoing long-term hemodialysis (HD) are known to have abnormal blood concentrations of antioxidant minerals; concurrent oxidative stress can contribute to increased vascular calcification. This study aims to evaluate the associations between circulating antioxidant minerals and clinical biomarkers of vascular calcification in HD patients. Blood biochemical parameters, antioxidant minerals (selenium (Se), zinc (Zn), copper (Cu), and magnesium (Mg)), and several promoters and inhibitors of calcification (matrix Gla protein (MGP), fibroblast growth factor-23 (FGF-23), matrix metalloproteinases (MMP-2 and -9), and tissue inhibitors of metalloproteinase (TIMP-1 and -2)) were determined in HD patients (n = 62) and age- and sex-matched healthy individuals (n = 30). Compared with healthy subjects, HD patients had significantly lower plasma concentrations of Se and Zn, increased Cu and Mg, and higher levels of oxidative stress and inflammatory markers (Cu/Zn ratios, malondialdehyde (MDA), advanced glycation end products (AGEs), and C-reactive protein (CRP)). We observed that HD patients had significantly lower concentrations of MGP and higher levels of FGF-23, MMP-2 and -9, TIMP-1 and -2, and MMP-2/TIMP-2 and MMP-9/TIMP-1 ratios. We also observed significant relationships between the concentrations of these minerals and calcification biomarkers in HD patients. These results suggest that changes in the homeostasis of antioxidant minerals (Se, Zn, Cu, and Mg) may contribute to the effects of oxidative stress and inflammatory status, thereby participating in the mechanism for accelerated vascular calcification in patients undergoing long-term HD.

Supplementation with Selenium yeast on the prooxidant-antioxidant activities and anti-tumor effects in breast tumor xenograft-bearing mice.

Selenium (Se) is essential for antioxidant activity involved in immune function and anti-carcinogenic action, whereas at higher concentrations, Se may have pro-oxidant properties. The present study was aimed at determining the effects of Se supplementation, as Se yeast, on oxidative stress in non-tumor/tumor tissues, as well as regulation of the apoptotic process, and immune responses in mice-bearing breast tumor xenografts. Female BALB/cByJNarl mice were divided into control (CNL and CNL-con), Se-supplemented control (CNL-HS, given as a single oral dose of 912 ng Se daily), breast tumor-bearing (TB and TB-con), TB-LS (228 ng Se), TB-MS (456 ng Se) and TB-HS (912 ng Se) groups. All mice were treated with/without Se for 14 days. A number of variables were further measured. Compared with the TB groups, tumor bearing mice with Se supplement had increased plasma Se concentrations, reduced erythrocyte Se-dependent glutathione peroxidase (GPx) activity and malondialdehyde (MDA) products and inhibited tumor growth. They have also higher Se concentrations in non-tumor and tumor tissues. Significantly elevated concentrations of MDA and reduced GPx activities, as well as increased anti-apoptotic bcl-2 and tumor suppressor p53 concentrations in tumor tissues were observed as Se accumulated in tumor, whereas lower MDA products were found in various non-tumor tissues than did the corresponding values. Further, there were elevated concentrations of Th1-derived cytokines and decreased Th2-type interleukin (IL)-4 in tumor-bearing mice with the treatment of Se. In conclusion, accumulation of Se in tumors may induce oxidative stress and p53-dependent pro-oxidative apoptosis, thus inhibiting the growth of breast tumor.

Effects of Selenium Yeast on Oxidative Stress, Growth Inhibition, and Apoptosis in Human Breast Cancer Cells.

Recent evidence suggests that selenium (Se) yeast may exhibit potential anti-cancer properties; whereas the precise mechanisms remain unknown. The present study was aimed at evaluating the effects of Se yeast on oxidative stress, growth inhibition, and apoptosis in human breast cancer cells. Treatments of ER-positive MCF-7 and triple-negative MDA-MB-231 cells with Se yeast (100, 750, and 1500 ng Se/mL), methylseleninic acid (MSA, 1500 ng Se/mL), or methylselenocysteine (MSC, 1500 ng Se/mL) at a time course experiment (at 24, 48, 72, and 96 h) were analyzed. Se yeast inhibited the growth of these cancer cells in a dose- and time-dependent manner. Compared with the same level of MSA, cancer cells exposure to Se yeast exhibited a lower growth-inhibitory response. The latter has also lower superoxide production and reduced antioxidant enzyme activities. Furthermore, MSA (1500 ng Se/mL)-exposed non-tumorigenic human mammary epithelial cells (HMEC) have a significant growth inhibitory effect, but not Se yeast and MSC. Compared with MSA, Se yeast resulted in a greater increase in the early apoptosis in MCF-7 cells as well as a lower proportion of early and late apoptosis in MDA-MB-231 cells. In addition, nuclear morphological changes and loss of mitochondrial membrane potential were observed. In conclusion, a dose of 100 to 1500 ng Se/mL of Se yeast can increase oxidative stress, and stimulate growth inhibitory effects and apoptosis induction in breast cancer cell lines, but does not affect non-tumorigenic cells.

Arsenic, cadmium, lead, and aluminium concentrations in human milk at early stages of lactation.

BACKGROUND: Human milk is considered to be the best nutrition for all infants because it provides the optimal source of nutritional, immunological, developmental, psychological, economic, practical, and environmental benefits in both the short and long terms. To the best of our knowledge, few studies in Taiwan have examined the toxicant levels in breast milk and associated factors. METHODS: The research was carried out over a 6-month period. Forty-five healthy lactating women, who delivered full-term newborns at our maternity ward, were recruited, and all participants had been living in coastal urban areas of mid-Taiwan for at least 3 years. One hundred and eighty human milk samples were collected on four occasions, which were classified into four lactation stages as follows: colostrums, transitional milk, early mature milk, and mature milk. RESULTS: We found that lead, cadmium, aluminium, and arsenic concentrations were the highest in colostrums: 13.22 ± 3.58 ng/mL, 1.37 ± 0.94 ng/mL, 56.45 ± 22.77 ng/mL, and 1.50 ± 1.50 ng/mL, respectively. The results of lead, cadmium, aluminium, and arsenic determination in human milk samples demonstrated a trend of decline of microelement concentrations with advancing stages of lactation. We found that the infants of smoking mothers were exposed to more cadmium than infants of nonsmoking mothers (p < 0.05). CONCLUSION: According to our findings, frequent routine sampling of breast milk is worthwhile. Prevention strategies including behavior modification and education on proper nutrition should be provided to women who are at high risk of toxicant exposure. In summary, breastfeeding is still generally encouraged and recommended.

Zinc supplementation alters plasma aluminum and selenium status of patients undergoing dialysis: a pilot study.

End stage renal disease patients undergoing long-term dialysis are at risk for abnormal concentrations of certain essential and non-essential trace metals and high oxidative stress. We evaluated the effects of zinc (Zn) supplementation on plasma aluminum (Al) and selenium (Se) concentrations and oxidative stress in chronic dialysis patients. Zn-deficient patients receiving continuous ambulatory peritoneal dialysis or hemodialysis were divided into two groups according to plasma Al concentrations (HA group, Al > 50 g/L; and MA group, Al > 30 to ≤ 50 g/L). All patients received daily oral Zn supplements for two months. Age- and gender-matched healthy individuals did not receive Zn supplement. Clinical variables were assessed before, at one month, and after the supplementation period. Compared with healthy subjects, patients had significantly lower baseline plasma Se concentrations and higher oxidative stress status. After two-month Zn treatment, these patients had higher plasma Zn and Se concentrations, reduced plasma Al concentrations and oxidative stress. Furthermore, increased plasma Zn concentrations were related to the concentrations of Al, Se, oxidative product malondialdehyde (MDA), and antioxidant enzyme superoxide dismutase activities. In conclusion, Zn supplementation ameliorates abnormally high plasma Al concentrations and oxidative stress and improves Se status in long-term dialysis patients.

Distribution of selenium and oxidative stress in breast tumor-bearing mice.

The present study investigated the effects of breast tumors on the blood and tissue distribution of essential trace mineral selenium (Se), and oxidative stress status of mice. Female 10-week-old BALB/cByJNarl mice were randomly assigned into control (CNL) and breast tumor-bearing (TB) groups. TB mice were injected subcutaneously into the right hind thigh with 5 × 10(6) EMT6 mouse mammary tumor cells. After 22 days, we measured Se concentrations, Se-dependent glutathione peroxidase (GPx) activities, and malondialdehyde (MDA) products (indicator of oxidative stress) in plasma, various tissues, and plasma vascular endothelial growth factor (VEGF) concentrations. There were no significant differences in body weights and daily intake between both groups. Compared with the CNL group, TB mice have decreases in plasma Se concentrations and GPx activities, as well as higher plasma VEGF and MDA concentrations. Plasma Se concentrations were also negatively correlated with plasma MDA and VEGF concentrations. Furthermore, tissue Se concentrations and GPx activities in TB animals were lower; whereas the MDA concentrations higher in various tissues including liver, kidney, brain, lung, spleen, and thymic tissues. In conclusion, disruption of Se homeostasis critically reflects oxidative stress in target tissues, thus may increase the risk for progression of breast cancer and metastasis.

The relationship of plasma aluminum to oxidant-antioxidant and inflammation status in asthma patients.

Aluminum (Al) is a non-essential mineral which human beings are exposed to on day-to-day life. The purpose of this study was to assess the concentrations of plasma Al and the relationship of those levels with risk factors for asthma. In total, 27 allergic asthmatics and 30 healthy volunteers were enrolled. Plasma Al and selected blood parameters were measured, and a pulmonary function test was performed. Higher Al concentrations were found in the asthmatics than the healthy controls. Increased immunoglobulin E, high-sensitivity C-reactive protein, lipid peroxidation products, and pro-inflammatory cytokines (tumor necrosis factor-α and interleukin [IL]-4) were observed, but IL-10 and overall antioxidant and enzyme activities were lower. Associations between oxidative-antioxidant status and inflammatory markers with plasma Al levels in asthmatics were noted. Al status was also linked to cytokine concentrations and pulmonary function. In conclusion, abnormal Al distribution may further precipitate oxidative stress and inflammation, alter Th1/Th2 lymphocyte balance, and therefore contribute to the development of asthma.

Plasma aluminum is a risk factor for oxidative stress and inflammation status in hemodialysis patients.

OBJECTIVES: The association between aluminum (Al), essential trace metals, oxidative stress, and inflammation status was evaluated in hemodialysis patients. DESIGN AND METHODS: Biochemical parameters in blood were determined in long-term hemodialysis patients (n=69) and age- and sex-matched healthy individuals (n=30). RESULTS: Compared with healthy subjects, patients had significantly higher concentrations of plasma Al. Elevated Al was negatively associated with the essential metals zinc, selenium, and iron. Al concentrations were strongly and positively correlated with contents of the oxidation products malondialdehyde and protein carbonyl. Inverse relationships were observed between Al concentrations and reduced concentrations of glutathione, ß-carotene, vitamin C, and vitamin E. Patients were also observed to have significantly increased production values of plasma high-sensitivity C-reactive protein, tumor necrosis factor-α, and interleukin-5. CONCLUSION: An increased plasma Al concentration is associated with disturbed concentrations of essential metals, increased oxidative stress, and increased inflammation status in hemodialysis patients.

Cu/Zn ratios are associated with nutritional status, oxidative stress, inflammation, and immune abnormalities in patients on peritoneal dialysis.

OBJECTIVES: We evaluated the relationship of the plasma copper/zinc (Cu/Zn) ratio with nutritional status, inflammation, oxidative stress, and immune function in peritoneal dialysis patients. DESIGN AND METHODS: Clinical and laboratory parameters were measured in patients (n=45) and age- and sex-matched healthy individuals (n=30). RESULTS: There were significant negative correlations of the Cu/Zn ratio with nutrition-related parameters (body mass index [BMI], creatinine, hemoglobin, and albumin) and antioxidant (vitamin C and E) levels and positive correlations of the Cu/Zn ratio with the levels of high sensitivity C-reactive protein (hs-CRP) and oxidation products (malondialdehyde [MDA] and protein carbonyl). The Cu/Zn ratio was negatively correlated with the percentages of B- and T-lymphocyte subsets and the ratio of CD4/CD8 antigens. CONCLUSIONS: In peritoneal dialysis patients, elevated Cu/Zn ratios are associated with malnutrition, increased oxidative stress, inflammation, and disrupted immune status.

Aluminum accumulation induced testicular oxidative stress and altered selenium metabolism in mice.

Present work was carried out to investigate how testicular selenium (Se) metabolisms respond to oxidative stress induced by aluminum (Al). Mice were intraperitoneally exposed to 0, 7, or 35mg Al/kg/d for 14 days (CNL, LAL and HAL groups). Al administration significantly increased Al, reactive oxygen radical and malondialdehyde (MDA) levels, as well as decreased glutathione peroxidase (GPx) and glutathione reductase (GR) activities in serum and testes. The serum concentrations of Se were remarkably lower at LAL and HAL groups compared to the controls, whereas the testicular Se levels significantly reduced only in the HAL group. In addition, RT-PCR analysis revealed an increased testicular selenoprotein P (SelP) expression by Al treatment. Western blot analysis showed increased levels of SelP protein expression in the LAL group, but the expression levels were significantly reduced in HAL group. It was suggested that altered metabolism of Se, further stimulated testicular SelP transcription that may compensate for the loss of SelP protein resulted from Al-induced oxidative damage.

Effects of peritoneal aluminum overload on polyamines and nitric oxide contents of testes and epididymis in the mice.

Polyamines are involved in cellular growth, differentiation and regulation of oxidative stress. The present investigation was to determine the effect of aluminum (Al) toxicity on the nitric oxide products (NO(x)) and metabolism of polyamines in mouse testes and epididymis. Aluminum chloride, AlCl(3,) was administered intraperitoneally to CD-1 adult male mice at dosages of 0, 7 or 35mg Al/kg body weight/day for 14 days (C, LAL and HAL groups). Results obtained show that the weights of epididymis in HAL animals are significantly decreased due to Al administration. Al treatment significantly induced higher Al concentrations in serum, testis and epididymis tissue. In addition, the serum and testicular and epididymal NO(x) production in HAL and testicular NO(x) in LAH groups were increased remarkably compared to the control animals. On the contrary, the contents of putrescine and spermine in testis were significantly lower than the values of controls and LAL groups. The epididymal spermine levels of HAL animal also decreased significantly. It is suggested that the polyamine biosynthesis in the mouse testis and epididymis can be affected by Al, which is associated with the NO(x) production in the male reproductive toxicity.

Aluminum-induced suppression of testosterone through nitric oxide production in male mice.

Excessive nitric oxide (NO) production in mice serum and testis due to aluminum (Al) exposure has been shown in previous studies. The aim of this study was to further investigate the role of NO on aluminum-suppressed testosterone level in male CD-1 mice. Each animal in six groups, was given intraperitoneal injections of either saline, aluminum chloride (AlCl(3)), l-N(6)-(1-iminoethyl) lysine (NO synthase inhibitor, l-NIL), or Al chloride along with l-NIL for a period of 12 days. These groups were denoted as C (control, saline), AL (35mg Al/kg/day, saline), NIL240 (total 240mg l-NIL/kg, saline), ALNIL240 (35mg Al/kg/day, total 240mg l-NIL/kg), ALNIL60 (35mg Al/kg/day, total 60mg l-NIL/kg), and NIL60 (total 60mg l-NIL/kg, saline). Results indicated that serum/testicular aluminum levels increased significantly in aluminum-treated animals compared to the controls, whereas the values observed from groups ALNIL240 than AL/ALNIL60 were markedly lower. Aluminum administration significantly increased NO production and decreased both testicular adenosine 3',5'-cyclic monophosphate (cAMP) and testosterone levels. A lower level of NO and higher concentrations of cAMP and testosterone observed in the ALNIL240 group indicated that the protective effect of NO synthase blockage was significant, although incomplete. In addition, aluminum induction significantly elevated the testicular cholesterol, but the values were lower in the ALNIL240 group than the AL or the ALNIL60 group. Finally, it was suggested that aluminum compounds exerted a significant adverse effects on the steroidogenesis and cAMP, which aided in the transport of cholesterol to the inner mitochondrial membrane. Furthermore, nitric oxide synthase blockage prevented aluminum-induced reproductive toxicity.

The influence of aluminum exposure on male reproduction and offspring in mice.

The dominant lethal assay was utilized to assess the reproductive performance in male mice, possible genetic hazards, and persistent damage of aluminum (Al). Al chloride, AlCl(3), was administered subcutaneously to CD-1 adult male mice at dosages of 0, 7, or 13mg Al/kg body weight/day for 2 weeks of pre-mating periods. Females were not dosed at any time during this study. At the end of the exposure period, each male was caged with three virgin females each day. The mean mating frequencies of the Al-treated groups reduced consistently from week 4 to 6, and a dramatic reduction in male fertility was also observed. However, the mating frequency restored to near normal control levels as the experiment terminated. Results showed significantly higher numbers of post-implantation losses, foetal mortality, and induced petechial haemorrhage; also significant decreases in body weights of viable foetus throughout weeks 3-8 in the Al-treated groups. The weights of the reproductive organs of the Al-dosed animals decreased significantly as Al accumulation increased in the testes. The spermatogenetic impairment within the seminiferous tubules was also apparent. Nevertheless, these disturbances disappeared at the end of the experiments. In summary, the results demonstrated that Al exerted substantial hazards on male reproductive function and produced genetic toxicity. However, these effects were found to be reversible.

Distribution patterns of trace metals and of lipid peroxidation in plasma and erythrocytes of rat exposed to aluminum.

Significant decreases of the hematocrit, hemoglobin, and plasma iron levels were observed in rats receiving daily intraperitoneal injections of aluminum at a dose of 27 mg Al/kg body wt for 3 wk, as compared to untreated controls. The activity of alkaline phosphatase was also significantly lower in the treated animals as a result of the accumulation of aluminum in the liver (p<0.05). Following aluminum administration, the plasma concentrations of aluminum and copper were also significantly increased, whereas the plasma zinc levels and oxidative stress measured through thiobarbituric acid reaction products showed nonsignificant differences between the two groups (p>0.05). The erythrocyte concentrations of aluminum, copper, zinc, and iron and of superoxide dismutase activity were found to be significantly higher in the study group as compared to controls. The treated animals also showed evidence of higher oxidative stress in comparison to controls. These results suggest that erythrocyte aluminum accumulation could result in abnormal trace element homeostasis and increasing oxidative stress, which might be a mechanism of aluminum-induced anemia.

Alteration of trace element distribution and testis ACE activity in mice with high peritoneal aluminum.

The present investigation was conducted to assess the effects of subacute aluminum (Al) exposure on testicular zinc (Zn), copper (Cu), and iron (Fe) distribution in mice. Animals were intraperitoneally exposed to 0, 13, or 35 mg Al/kg body weight/d for a period of 14 d. Al concentrations in serum and testis in Al-treated animals were significantly higher than those of controls. The serum concentrations of Fe were lower, whereas serum Zn and Cu showed a pattern comparable to that of controls. The accumulation of testicular Fe and Cu remarkably increased in Al-exposed groups, whereas the Zn concentration in testis was significantly reduced only at the highest dose of Al exposure. The values of testicular thiobarbituric acid reactive substances (TBARS) were also increased after Al administration, indicating increased lipid peroxidation and oxidative stress. In addition, when the testicular Al was increased, the testis-specific angiotensin-converting enzyme (testis ACE) was noted. The results of this study indicated that part of the effect of Al intoxication on testis might contribute to abnormal metabolism of other minerals, such as Fe, Zn, and/or Cu. It was also suggested that reduced testis ACE activity presumably plays an important role in oxidative damage of Al-induced testicular toxicity.