Qiang Niu

Toxic effect of PBDE-47 on thyroid development, learning, and memory, and the interaction between PBDE-47 and PCB153 that enhances toxicity in rats

Polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) are widespread environmental contaminants. There are potential interactive effects between PBDEs and PCBs, as these compounds share similar structures. The developmental neurotoxicity of 2, 20, 4, 40-tetrabromodiphenyl ether (PBDE-47) and the interaction of PBDE-47 with 2, 20, 4, 40, 5, 50-hexachlorobipheny (PCB153) were investigated herein, as the dominant congener forms of PBDEs and PCBs, respectively. SD rats were exposed to a single oral dose of PBDE-47 (1, 5, and 10 mg/g) and/or PCB153 (5 mg/g) on post-natal day (PND) 10. Concentrations of PBDE47, triiodothyronine (T3), thyroxine (T4), and thyroid-stimulating hormone (TSH) in serum; organ-to-body weight ratios; as well as long-term learning and memory were measured in 2-month-old rats. The present study found that some doses of PBDE-47 decreased the organ-to-body weight ratios of the thyroid and uterus, decreased the concentration of T4 in serum, and increased the organ-to-body weight ratio of the ovaries (p < 0.05). PCB153 could increase the action of PBDE-47 during combined exposure, but this interaction was not found between PBDE-47 and PCB153. In a Morris water maze experiment, the latency periods were significantly prolonged and time ratios were obviously depressed in all PBDE-47-treated groups compared to the control (p < 0.05); furthermore, significant interactions between PBDE-47 and PCB153 were observed (p < 0.05). In conclusion, PBDE-47 may depress thyroid development as well as the long-term learning and memory capabilities in adult rats exposed to PBDE-47 on PND 10. PCB153 can interact with PBDE-47, resulting in an increase in developmental neurotoxicity.

Mechanisms underlying the developmental neurotoxic effect of PBDE-47 and the enhanced toxicity associated with its combination with PCB153 in rats.

To explore the mechanisms underlying the developmental neurotoxic effect of PBDE-47 and its interaction with PCB153, expression levels of mRNA and proteins of the x-chromosome-linked inhibitor of apoptosis protein (XIAP), death associated protein kinase (DAPK), caspase3, caspase12 and cytochrome C in the hippocampus of 2-month-old rats exposed to a single oral dose of PBDE-47 and/or PCB153 on post natal day (PND) 10 were examined. Four levels of PBDE-47 (0, 1, 5, 10 mg/kg) and two levels of PCB153 (0 and 5mg/kg) were added to corn oil in a 4 x 2 factorial completely randomized design study. Meanwhile, the ultrastructures of neurons in the hippocampal CA1 region were observed and the learning and memory capacities were measured in these rats. The results suggested that the mRNA and protein expression levels of all examined genes (with the exception of cytochrome C mRNA in female rats) were significantly changed at some doses (P<0.05); additionally, the total distance swam by rats to reach an escape platform was significantly increased and the ratio of distance taken in the platform quadrant to total distance was notably decreased in all treated groups in the water maze experiment (P<0.05) compared to the control. Numerous alterations were observed in the ultrastructure of neurons in PBDE-47 alone or combination of PBDE-47 and PCB153 groups. Furthermore, an interaction was found between PBDE-47 and PCB153 in lengthening the total distance taken to the platform and decreasing the platform quadrant ratios in the water maze experiment, as well as in the inducing of caspase3, caspase12 and cytochrome C mRNA and protein expression (with exception of cytochrome C mRNA in female rats) in the hippocampus. We conclude that PBDE-47 may induce developmental neurotoxicity in rats via three classic apoptosis pathways, and it may interact with PCB153 to enhance developmental neurotoxicity.

Effects of the Fas/Fas‐L pathway on fluoride‐induced apoptosis in SH‐SY5Y cells

The mechanisms underlying fluoride‐induced apoptosis in neurons still remain unknown. To investigate apoptosis, caspase‐3 activity, and mRNA expression of Fas, Fas‐L, and caspases (‐3 and ‐8) induced by fluoride, human neuroblastoma (SH‐SY5Y) cells were incubated with 0, 20, 40, and 80 mg/L sodium fluoride (NaF) for 24 h in vitro. The data show that cell viability in the 40 and 80 mg/L fluoride groups were significantly lower than that of the control group. The percentages of apoptosis in the 40 and 80 mg/L fluoride groups were markedly higher than those in the control group, and they increased with the increase in fluoride concentration. The activity of caspase‐3 and mRNA expression levels for Fas, Fas‐L, and caspases (‐3 and ‐8) in the 40 and 80 mg/L fluoride groups were significantly higher than those in the control group. An agonistic anti‐Fas monoclonal antibody (CH‐11) significantly augmented apoptosis induction by fluoride, showing a synergistic effect, while a Fas‐blocking antibody (ZB4) partly inhibited fluoride‐induced apoptosis of SH‐SY5Y cells. The results indicate that fluoride exposure could induce apoptosis in SH‐SY5Y cells, and the Fas/Fas‐L signaling pathway may play an important role in the process.

Relationship between intracellular Ca2+ and ROS during fluoride‐induced injury in SH‐SY5Y cells

The mechanisms underlying the neurotoxicology of endemic fluorosis still remain obscure. To explore lactate dehydrogenase (LDH) leakage, intracellular Ca2+ concentration ([Ca2+]i) and reactive oxygen species (ROS) production induced by fluoride, human neuroblastoma (SH‐SY5Y) cells were incubated with sodium fluoride (NaF, 20, 40, 80 mg/L) for 24 h, with 40 mg/L NaF for 3, 6, 12, 18, 24 h, and N‐acetyl‐L‐cysteine (NAC), ethyleneglycol‐bis‐(β‐aminoethyl ether)‐N,N,N′,N′‐tetraacetic acid (EGTA), 1,2‐bis(O‐aminophenoxy)ethane‐N,N,N′,N′‐tetraacetic acid tetra(acetoxymethyl) ester (BAPTA‐AM) alone or combined with fluoride (40 mg/L) respectively for 12 h in vitro. The results showed that the LDH levels in the 40 and 80 mg/L fluoride‐treated groups were significantly higher than that of the control group (in the test level of 0.05, the difference were statistical significance). [Ca2+]i and ROS reached a peak at 3 h and 12 h respectively after exposure to 40 mg/L fluoride. Fluoride coincubated with NAC (antioxidant) dramatically decreased ROS and LDH levels compared with the fluoride only group (in the test level of 0.05, the difference were statistical significance). However, fluoride‐induced increase in [Ca2+]i was not affected by NAC. BAPTA‐AM (intracellular calcium chelator) markedly lowered fluoride‐induced increase of [Ca2+]i, ROS and LDH levels while EGTA (extracellular calcium chelator) have no effects on them. These results indicate that fluoride‐related Ca2+ release from the site of intracellular calcium storage causes the elevation of ROS contributing to the cytotoxicity in SH‐SY5Y cells.

ERK1/2-mediated disruption of BDNF–TrkB signaling causes synaptic impairment contributing to fluoride–induced developmental neurotoxicity

Excessive exposure to fluoride has adverse effects on neurodevelopment, but the mechanisms remain unclear. This study aimed to investigate the effects of fluoride exposure on synaptogenesis, and focused on the role of brain-derived neurotrophic factor (BDNF)-tyrosine kinase B (TrkB) signaling in these effects. Using Sprague-Dawley rats developmentally exposed to sodium fluoride (NaF) from pregnancy until 6 months of delivery as in vivo model, we showed that fluoride impaired the cognitive abilities of offspring rats, decreased the density of dendritic spines and the expression of synapse proteins synaptophysin (SYN) and postsynaptic density protein-95 (PSD-95) in hippocampus, suggesting fluoride-induced cognitive deficit associates with synaptic impairment. Consistently, NaF treatment reduced dendritic outgrowth and expression of SYN and PSD-95 in human neuroblastoma SH-SY5Y cells. Further studies demonstrated that the BDNF-TrkB axis was disrupted in vivo and in vitro, as manifested by BDNF accumulation and TrkB reduction. Importantly, fluoride treatment increased phospho-extracellular signal-regulated kinases 1 and 2 (p-ERK1/2) expression, while inhibition of p-ERK1/2 significantly attenuated the effects of NaF, indicating a regulating role of p-ERK1/2 in BDNF-TrkB signaling disruption. Collectively, these data suggest that the developmental neurotoxicity of fluoride is associated with the impairment of synaptogenesis, which is caused by ERK1/2-mediated BDNF-TrkB signaling disruption.