TDCPP(三(1,3-二氯异丙基)磷酸酯)是磷酸三(2,3-二溴丙基)酯(Tris)的氯化类似物,它是环境中检测到最多的有机磷阻燃剂(OPFR)之一。
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TDCPP is a chlorinated analog of tris(2,3-dibromopropyl)phosphate (Tris) which is one of the most detected organophosphorus flame retardants (OPFRs) in the environment.
Exposure to TDCPP does not significantly affect cell viability until at concentration >68 μg/mL. HCECs show a 16% cell viability loss after exposing to 136 μg/mL TDCPP. Moreover, TDCPP induces a sharp decrease in viable cells (87%) after exposing to ≥272 μg/mL TDCPP. Based on cell viability, the LC50 value for TDCPP is 202 μg/mL using a nonlinear regression. Compare to controls, TDCPP-exposed cells exhibit a concentration-dependent increase in apoptosis. Anti-apoptotic Bcl-2 protein expression is increased to 1.4 fold after exposing to 2 μg/mL TDCPP, 1.2-folds at 20 μg/mL but dynamically decreased to 0.4 fold at 200 μg/mL compare to control. The caspase-3 activity is increased to 2.1 folds of the control at 200 μg/mL TDCPP[1]. TDCPP inhibits cell growth at lower concentrations (IC50 of 27 μM), while cell viability and toxicity are affected at higher concentrations (IC50 of 171 μM and 168 μM, respectively)[2].
[1]. Xiang P, et al. Effects of organophosphorus flame retardant TDCPP on normal human corneal epithelial cells: Implications for human health. Environ Pollut. 2017 Nov;230:22-30. [2]. Killilea DW, et al. Flame retardant tris(1,3-dichloro-2-propyl)phosphate (TDCPP) toxicity is attenuated by N-acetylcysteine in human kidney cells. Toxicol Rep. 2017 May 17;4:260-264.
Kinase experiment: |
The cellular ATP contents are determined in HCECs grown in DMEM containing 0, 2, 20, or 200 μg/mL TDCPP using a luciferase-based ATP assay kit according to the manufacturer's guideline. Briefly, after 24 h exposure, HCECs are lysed with lysis buffer. Lysates are then centrifuged at 12,000 g at 4°C for 5 min. Then, 100 μL of supernatant is mixed with 100 μL ATP detection working dilution. Luminance is examined by an fluorescence microplate reader[1]. |
Cell experiment: |
To examine the effects of TDCPP on cell viability, HCECs are planted into 96-well plate (100 μL/well) at density of 1×105 cells/mL overnight. Then, the medium is changed into fresh medium containing 0.034, 0.34, 3.4, 34, 68, 136, 272, or 340 μg/mL of TDCPP and solvent vehicle (0.1%, v/v) and incubated for 24 h. Cell viability is detected using CCK-8 cell viability assay kit according to the manufacturer's instructions. After exposure, cellular morphology is observed and recorded by an inverted microscopy[1]. |
参考文献: [1]. Xiang P, et al. Effects of organophosphorus flame retardant TDCPP on normal human corneal epithelial cells: Implications for human health. Environ Pollut. 2017 Nov;230:22-30. |
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