Comparative assessment of neurotoxicity impacts induced by alkyl tri-n-butyl phosphate and aromatic tricresyl phosphate in PC12 cells.

Organophosphate flame retardants (OPFRs) have become a growing concern due to their potential environmental and health risk. However, limited studies have described the toxicity, particularly neurotoxicity of alkyl and aromatic OPFRs. This study investigated the neurotoxicity of alkyl tri-n-butyl phosphate (TnBP) and aromatic tricresyl phosphate (TCP) to rat adrenal pheochromocytoma (PC12) cells for 24 h. Viability detection showed dose-response toxicity effect of TCP and TnBP to PC12 cells. The half-maximal inhibitory concentration of 24 h (24 h-IC50 ) of TCP and TnBP were 2415.61 and 338.09 µM, respectively. Both TnBP and TCP significantly changed the acetylcholinesterase (AChE) activity, and TnBP is more likely to cause neurotoxicity to PC12 cells compared to TCP. Also, The results of LDH and caspase-3 activity detection as well as Hoechst staining suggested that cell apoptosis induced by TCP and TnBP may be the primary pathway. These findings provide a toxicity data of aromatic and alkyl-substituted OPFRs to PC12 cells, and a new insight into the toxicity of OPFRs on health risk assessment.

Intestinal damage, neurotoxicity and biochemical responses caused by tris (2-chloroethyl) phosphate and tricresyl phosphate on earthworm.

Organophosphate esters (OPEs) draw growing concern about characterizing the potential risk on environmental health due to its wide usage and distribution. Two typical types of organophosphate esters (OPEs): tris (2-chloroethyl) phosphate (TCEP) and tricresyl phosphate (TCP) were selected to evaluate toxicity of OPEs to the soil organism like earthworm (Eisenia fetida). Histopathological examination (H&E), oxidative stress, DNA damage and RT-qPCR was used to identify the effects and potential mechanism of their toxicity. Hameatoxylin and eosin (H&E) demonstrated that intestinal cells suffered serious damage, and the observed up-regulation of chitinase and cathepsin L in mRNA levels confirmed it. Both TCEP and TCP significantly increased the DNA damage when the concentrations exceeded 1 mg/kg (p < 0.01), and a dose-response relationship was observed. In addition, TCEP and TCP also changed the acetylcholinesterase (AChE) activity and expression of genes associated with neurotoxic effects in earthworms even under exposure to low OPEs concentration (0.1 mg/kg). Moreover, genes associated with nicotinic acetylcholine receptors (nAChR) and carrier protein further demonstrated that highest concentration of TCEP (10 mg/kg) may have an overloading impact on the cholinergic system of E. fetida. Integrated Biological Response index (IBRv2) showed that TCEP exerted stronger toxicity than TCP under the same concentrations. We deduced that the observed intestinal damage, oxidative stress and neurotoxic effect might be the primary mechanisms of TCEP and TCP toxicity. This study provides insight into the toxicological effects of OPEs on earthworm model, and may be useful for risk assessment of OPEs on soil ecosystems.

Liver immune and lipid metabolism disorders in mice induced by triphenyl phosphate with or without high fructose and high fat diet.

Organophosphorus flame retardants (OPFRs) are frequently detected in food and human samples, and epidemiological studies have found that human exposure to aryl-OPFRs (triphenyl phosphate, TPP) is associated with lipid metabolism. Although toxicity studies suggest a potential obesity risk from TPP exposure, the molecular mechanism remains unclear. This study investigated the subchronic dietary effects on mouse liver significantly changed proteins (SCPs) and elucidated the underlying molecular mechanisms of TPP with or without a high-fructose and high-fat (HFF) diet. Male C57BL/6J mice were exposed to low-dose TPP (corresponding to the oral reference dose, 10 µg/kg body weight (bw)/day) and high-dose TPP (1000 µg/kg bw/day) for 12 weeks. The results showed that exposure to TPP generated changes of liver function and organelle damage as well as increases in total cholesterol and triglyceride levels. TPP exposure at a low dose damaged the liver immune system via major histocompatibility complex-related proteins involved in antigen processing and presentation. TPP exposure at a high dose caused disorders of the biosynthesis of unsaturated fatty acids and steroid hormones, thereby inducing lipid accumulation in the liver. Although 10 µg/kg TPP did not cause serious lipid metabolism disorders in the liver, significant overexpression of fatty acid-binding protein 5, malic enzyme 1, and other related SCPs was observed, which led to disorders of cholesterol metabolism and lipogenesis to activate the proliferator-activated receptor signaling pathway and thus induced potential obesity risks. In addition, lipid metabolism disorders related to TPP were aggravated under the HFF diet, impairing liver mitochondrial and endoplasmic reticulum function in mice by altering the activity of cytochrome P450 enzyme subfamilies. These findings provide an in-depth understanding of the molecular toxicity mechanisms and health risks associated with subchronic exposure to TPP under different dietary regimes.

Triphenyl phosphate exposure induces kidney structural damage and gut microbiota disorders in mice under different diets.

Exposure of humans to organophosphate flame retardants (OPFRs) and the consequent health risk have increased owing to the latter's widespread application. Although triphenyl phosphate (TPP), an OPFR, is a potential chemical determinant of liver function damage, its effects on kidney function in mice under high fructose/fat (HFF) diet are still unclear. In this study, C57BL/6J mice were fed HFF to generate an obesity model and mice were exposed to low dose (0.01 mg/kg/day; TPP-L) and high dose (1 mg/kg/day; TPP-H) of TPP for 12 weeks. Results showed that TPP-L and TPP-H combined with HFF, as well as TPP-H alone, caused kidney structural damage and gut microbiota disorders in mice. Inflammatory response induced by nuclear factor kappa B (NF-κB p65)/nod-like receptor protein 3 (NLRP3) and caspase-3 promoted kidney structure damage, as well as accumulation of triglyceride and total cholesterol and the protein residues in urine. Although TPP-L did not cause obvious structural damage in the kidneys, 0.01 mg/kg TPP induced significant inflammation and gut microbiota disorders. These findings provide new insights regarding health risk assessment after chronic exposure to TPP and HFF alone, as well as a combination of TPP with HFF in mice.

Toxicological effects of polystyrene microplastics on earthworm (Eisenia fetida).

Microplastics are plastic fragments of particle sizes less than 5 mm, which are widely distributed in marine and terrestrial environments. In this study, earthworms Eisenia fetida were exposed to 100 and 1000 µg of 100 nm and 1300 nm fluorescent polystyrene microplastics (PS-MPs) per kg of artificial soil for 14 days. Uptake or accumulation of PS-MPs in earthworm intestines, histopathological changes, oxidative stress, and DNA damage were assessed to determine the toxicological effects of PS-MPs on E. fetida. The results showed that the average accumulated concentrations in the earthworm intestines were higher for 1300 nm PS-MPs (0.084 ± 0.005 and 0.094 ± 0.003 µg/mg for 100 and 1000 µg/kg, respectively) than for 100 nm PS-MPs (0.015 ± 0.001 and 0.033 ± 0.002 µg/mg for 100 and 1000 µg/kg, respectively). In addition, histopathological analysis indicated that the intestinal cells were damaged after exposure to PS-MPs. Furthermore, PS-MPs significantly changed glutathione (GSH) level and superoxide dismutase (SOD) activity. The GSH levels were 86.991 ± 7.723, 165.436 ± 4.256-167.767 ± 18.642, and 93.590 ± 4.279-173.980 ± 15.523 µmol/L in the control, 100 nm, and 1300 nm PS-MPs treatment groups. In addition, the SOD activities were 10.566 ± 0.621, 9.039 ± 0.787-9.408 ± 0.493, and 7.959 ± 0.422-9.195 ± 0.327 U/mg protein for the control, 100 nm, and 1300 nm PS-MPs treatment groups, respectively, indicating that oxidative stress was induced after PS-MPs exposure. Furthermore, the comet assay suggested that exposure to PS-MPs induced DNA damage in earthworms. Overall, 1300 nm PS-MPs showed more toxic effect than 100 nm PS-MPs on earthworms. These findings provide new insights regarding the toxicological effects of low concentrations of microplastics on earthworms, and on the ecological risks of microplastics to soil animals.