Toxics, Vol. 10, Pages 698: Triflumizole Induces Developmental Toxicity, Liver Damage, Oxidative Stress, Heat Shock Response, Inflammation, and Lipid Synthesis in Zebrafish

Figure 1. Exposure patterns and the survival percentage of zebrafish after triflumizole exposure. (A) Three dpf embryos were exposed to 1, 2, and 3 mg/L triflumizole for 48 h, (B) Six dpf larvae were exposed to 0.5, 1, and 1.5 mg/L triflumizole for 24 h. (C) The percentage of survival of 3 dpf embryos after triflumizole exposure for 48 h. (D) The percentage of survival of 6 dpf larvae after triflumizole exposure for 24 h. dpf: d post-fertilization; LC50: lethal concentration 50.

Figure 1. Exposure patterns and the survival percentage of zebrafish after triflumizole exposure. (A) Three dpf embryos were exposed to 1, 2, and 3 mg/L triflumizole for 48 h, (B) Six dpf larvae were exposed to 0.5, 1, and 1.5 mg/L triflumizole for 24 h. (C) The percentage of survival of 3 dpf embryos after triflumizole exposure for 48 h. (D) The percentage of survival of 6 dpf larvae after triflumizole exposure for 24 h. dpf: d post-fertilization; LC50: lethal concentration 50.

Toxics 10 00698 g001

Figure 2. Effects of development after triflumizole exposure in zebrafish. (A) 3 dpf zebrafish embryos with morphological changes after triflumizole exposure for 48 h. (B) 6 dpf zebrafish larvae with liver degeneration after triflumizole exposure for 24 h. Blue arrows and red arrows indicate pericardial edema and liver degeneration, respectively. H: heart, the green dotted line; L: liver, the red dotted line; I: intestine, the blue dotted line; Y: yolk sac, the yellow dotted line. Scale bars = 500 μm; Scale bars = 100 μm.

Figure 2. Effects of development after triflumizole exposure in zebrafish. (A) 3 dpf zebrafish embryos with morphological changes after triflumizole exposure for 48 h. (B) 6 dpf zebrafish larvae with liver degeneration after triflumizole exposure for 24 h. Blue arrows and red arrows indicate pericardial edema and liver degeneration, respectively. H: heart, the green dotted line; L: liver, the red dotted line; I: intestine, the blue dotted line; Y: yolk sac, the yellow dotted line. Scale bars = 500 μm; Scale bars = 100 μm.

Toxics 10 00698 g002

Figure 3. Effects of triflumizole on enzyme activities in zebrafish. For 3 dpf zebrafish embryos, SOD activity (A), CAT activity (B), MDA content (C), and ALT activity (D) after triflumizole exposure for 48 h. For 6 dpf zebrafish larvae, SOD activity (E), CAT activity (F), MDA content (G), and ALT activity (H) after triflumizole exposure for 24 h. Data are expressed as the mean of three replicates ± standard error (SEM). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, * p < 0.05, ** p < 0.01). SOD: superoxide dismutase; CAT: catalase; MDA: malondialdehyde; ALT: alanine transaminase.

Figure 3. Effects of triflumizole on enzyme activities in zebrafish. For 3 dpf zebrafish embryos, SOD activity (A), CAT activity (B), MDA content (C), and ALT activity (D) after triflumizole exposure for 48 h. For 6 dpf zebrafish larvae, SOD activity (E), CAT activity (F), MDA content (G), and ALT activity (H) after triflumizole exposure for 24 h. Data are expressed as the mean of three replicates ± standard error (SEM). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, * p < 0.05, ** p < 0.01). SOD: superoxide dismutase; CAT: catalase; MDA: malondialdehyde; ALT: alanine transaminase.

Toxics 10 00698 g003

Figure 4. The expression of heat shock response genes (including hsp70, grp78, hsp90, and grp94) in zebrafish. (A) For 3 dpf zebrafish embryos, the expression of hsp70, grp78, hsp90, and grp94 after triflumizole exposure for 48 h. (B) For 6 dpf zebrafish larvae, the expression of hsp70, grp78, hsp90, and grp94 after triflumizole exposure for 48 h. Data are expressed as the mean of three replicates ± standard error (SEM). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, * p < 0.05, ** p < 0.01). hsp70: heat shock protein 70; grp78: glucose-regulated protein 78; hsp90: heat shock protein 90; grp94: glucose-regulated protein 94.

Figure 4. The expression of heat shock response genes (including hsp70, grp78, hsp90, and grp94) in zebrafish. (A) For 3 dpf zebrafish embryos, the expression of hsp70, grp78, hsp90, and grp94 after triflumizole exposure for 48 h. (B) For 6 dpf zebrafish larvae, the expression of hsp70, grp78, hsp90, and grp94 after triflumizole exposure for 48 h. Data are expressed as the mean of three replicates ± standard error (SEM). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, * p < 0.05, ** p < 0.01). hsp70: heat shock protein 70; grp78: glucose-regulated protein 78; hsp90: heat shock protein 90; grp94: glucose-regulated protein 94.

Toxics 10 00698 g004

Figure 5. The expression of inflammatory genes (including tnfα, il-1β, p65-nfκb, cox2a, and cox2b) in zebrafish. (A) For 3 dpf zebrafish embryos, the expression of tnfα, il-1β, p65-nfκb, cox2a, and cox2b after triflumizole exposure for 48 h. (B) For 6 dpf zebrafish larvae, the expression of tnfα, il-1β, p65-nfκb, cox2a, and cox2b after triflumizole exposure for 24 h. Data are expressed as the mean of three replicates ± standard error (SEM). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, ** p < 0.01). tnfα: tumor necrosis factor α; il-1β: interleukin 1, beta; p65-nfκb: p65-nuclear transcription factor κB; cox2a: cyclooxygenase type 2 a; cox2b: cyclooxygenase type 2 b.

Figure 5. The expression of inflammatory genes (including tnfα, il-1β, p65-nfκb, cox2a, and cox2b) in zebrafish. (A) For 3 dpf zebrafish embryos, the expression of tnfα, il-1β, p65-nfκb, cox2a, and cox2b after triflumizole exposure for 48 h. (B) For 6 dpf zebrafish larvae, the expression of tnfα, il-1β, p65-nfκb, cox2a, and cox2b after triflumizole exposure for 24 h. Data are expressed as the mean of three replicates ± standard error (SEM). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, ** p < 0.01). tnfα: tumor necrosis factor α; il-1β: interleukin 1, beta; p65-nfκb: p65-nuclear transcription factor κB; cox2a: cyclooxygenase type 2 a; cox2b: cyclooxygenase type 2 b.

Toxics 10 00698 g005

Figure 6. The expression of lipid synthesis genes (including srebp1, fas, acc, and ppar-γ) in zebrafish. (A) For 3 dpf zebrafish embryos, the expression of srebp1, fas, acc, and ppar-γ after triflumizole exposure for 48 h. (B) For 6 dpf zebrafish larvae, the expression of srebp1, fas, acc, and ppar-γ after triflumizole exposure for 48 h. Data are expressed as the mean of three replicates ± standard error (SEM). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, * p < 0.05, ** p < 0.01). srebp1: sterol regulatory element binding transcription protein 1; fas: fas cell surface death receptor; acc: acetyl-coa carboxylase; ppar-γ: peroxisome proliferator-activated receptor gamma.

Figure 6. The expression of lipid synthesis genes (including srebp1, fas, acc, and ppar-γ) in zebrafish. (A) For 3 dpf zebrafish embryos, the expression of srebp1, fas, acc, and ppar-γ after triflumizole exposure for 48 h. (B) For 6 dpf zebrafish larvae, the expression of srebp1, fas, acc, and ppar-γ after triflumizole exposure for 48 h. Data are expressed as the mean of three replicates ± standard error (SEM). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, * p < 0.05, ** p < 0.01). srebp1: sterol regulatory element binding transcription protein 1; fas: fas cell surface death receptor; acc: acetyl-coa carboxylase; ppar-γ: peroxisome proliferator-activated receptor gamma.

Toxics 10 00698 g006

Figure 7. Effects of the fish intestinal cell line after triflumizole exposure for 24 h. (A) The cell viability. (B) The expression of hsp70, grp78, hsp90, and grp94 after triflumizole exposure. (C) The expression of p65-nfκb, p105-nfκb, and cox2 after triflumizole exposure. (D) The expression of srebp1, lpl, ppar-γ and ppar-α after triflumizole exposure. Data are expressed as the mean of three replicates ± standard error (SEM). Values without a common superscript letter differ (p < 0.05, Tukey’s test). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, * p < 0.05, ** p < 0.01). Data are expressed as the mean of three replicates ± standard error (SEM). p65-nfκb: p65-nuclear transcription factor κB; p105-nfκb: p105-nuclear transcription factor κB; cox2: cyclooxygenase 2; lpl: lipoprotein lipase; ppar-α: peroxisome proliferator-activated receptor alpha.

Figure 7. Effects of the fish intestinal cell line after triflumizole exposure for 24 h. (A) The cell viability. (B) The expression of hsp70, grp78, hsp90, and grp94 after triflumizole exposure. (C) The expression of p65-nfκb, p105-nfκb, and cox2 after triflumizole exposure. (D) The expression of srebp1, lpl, ppar-γ and ppar-α after triflumizole exposure. Data are expressed as the mean of three replicates ± standard error (SEM). Values without a common superscript letter differ (p < 0.05, Tukey’s test). Asterisks denote significant differences between the control group and TFZ groups (determined by Dunnett post hoc comparison, * p < 0.05, ** p < 0.01). Data are expressed as the mean of three replicates ± standard error (SEM). p65-nfκb: p65-nuclear transcription factor κB; p105-nfκb: p105-nuclear transcription factor κB; cox2: cyclooxygenase 2; lpl: lipoprotein lipase; ppar-α: peroxisome proliferator-activated receptor alpha.

Toxics 10 00698 g007

留言 (0)

沒有登入
gif