Table 1.
Detection of biochemical parameters in the muscle and gill tissue for C60 NPs-exposed zebrafish after 12-day exposure.
| Biomarker | Control | C60 (1ppm) | C60 (2ppm) | Unit | Significance | ANOVA F value | p value |
|---|---|---|---|---|---|---|---|
| Muscle | |||||||
| ROS | 4.643 ± 0.356 a | 5.353 ± 0.586 a | 15.710 ± 1.881 b | U/μg total protein | YES | F (2, 6) = 28.740 | p = 0.0008 |
| 4-HNE | 1.028 ± 0.101 | 1.192 ± 0.223 | 1.005 ± 0.182 | U/μg total protein | NO | F (2, 6) = 0.341 | p = 0.7272 |
| MDA | 0.163 ± 0.011 | 0.096 ± 0.016 | 0.143 ± 0.021 | ng/μg total protein | NO | F (2, 6) = 4.416 | p = 0.0662 |
| TBARS | 3.317 ± 0.045 a | 3.922 ± 0.541 a | 20.240 ± 2.066 b | ng/μg total protein | YES | F (2, 6) = 60.600 | p = 0.0001 |
| catalase | 1.157 ± 0.003 a | 1.662 ± 0.200 a | 0.499 ± 0.045 b | U/μg total protein | YES | F (2, 6) = 24.300 | p = 0.0013 |
| SOD | 3.443 ± 0.199 a | 4.825 ± 0.787 a | 1.004 ± 0.204 b | U/μg total protein | YES | F (2, 6) = 16.050 | p = 0.0039 |
| ssDNA | 0.717 ± 0.007 a | 0.791 ± 0.162 a | 3.702 ± 0.229 b | U/μg total protein | YES | F (2, 6) = 110.600 | p < 0.0001 |
| catecholamine | 4.031 ± 0.063 a | 3.793 ± 0.732 a | 15.240 ± 1.847 b | ng/μg total protein | YES | F (2, 6) = 32.480 | p = 0.0006 |
| cortisol | 11.700 ± 0.413 a | 35.090 ± 5.455 b | 43.020 ± 5.051 b | pg/μg total protein | YES | F (2, 6) = 14.340 | p = 0.0052 |
| TNF-α | 5.877 ± 0.076 a | 5.603 ± 1.080 a | 19.880 ± 2.414 b | pg/μg total protein | YES | F (2, 6) = 28.600 | p = 0.0009 |
| IL1β | 0.406 ± 0.010 a | 0.365 ± 0.071 a | 2.651 ± 0.242 b | ng/μg total protein | YES | F (2, 6) = 80.890 | p < 0.0001 |
| Hif-1α | 13.880 ± 0.738 | 21.880 ± 4.270 | 19.170 ± 2.221 | pg/μg total protein | NO | F (2, 6) = 2.095 | p = 0.2042 |
| creatine kinase | 2.092 ± 0.019 a | 2.037 ± 0.412 a | 5.300 ± 0.473 b | pg/μg total protein | YES | F (2, 6) = 26.600 | p = 0.0010 |
| ATP | 293.400 ± 8.753 a | 257.900 ± 57.280 a | 89.440 ± 15.460 b | ng/μg total protein | YES | F (2, 6) = 9.909 | p = 0.0126 |
| creatinine | 5.715 ± 0.133 | 3.916 ± 1.222 | 5.543 ± 0.435 | U/μg total protein | NO | F (2, 6) = 1.738 | p = 0.2538 |
| metallothionine | 13.840 ± 0.537 | 13.470 ± 0.727 | 12.610 ± 0.564 | pg/μg total protein | NO | F (2, 6) = 1.055 | p = 0.4049 |
| Gill | |||||||
| ROS | 2.923 ± 0.193 a | 3.386 ± 0.644 a | 7.235 ± 1.219 b | U/μg total protein | YES | F (2, 6) = 8.675 | P = 0.0170 |
| 4-HNE | 0.552 ± 0.115 a | 1.048 ± 0.030 b | 0.792 ± 0.070 a,b | U/μg total protein | YES | F (2, 6) = 9.693 | p = 0.0132 |
| MDA | 0.123 ± 0.007 a | 0.114 ± 0.012 a | 0.024 ± 0.006 b | ng/μg total protein | YES | F (2, 6) = 39.050 | p = 0.0004 |
| TBARS | 2.504 ± 0.074 a | 3.135 ± 0.534 a | 6.630 ± 1.230 b | ng/μg total protein | YES | F (2, 6) = 8.213 | p = 0.0191 |
| Catalase | 0.875 ± 0.044 a | 0.815 ± 0.052 a | 0.481 ± 0.022 b | U/μg total protein | YES | F (2, 6) = 26.470 | p = 0.0011 |
| SOD | 2.501 ± 0.071 a | 2.946 ± 0.473 a | 1.088 ± 0.082 b | U/μg total protein | YES | F (2, 6) = 11.990 | p = 0.0080 |
| ssDNA | 0.522 ± 0.018 a | 0.941 ± 0.056 a | 1.555 ± 0.204 b | U/μg total protein | YES | F (2, 6) = 17.930 | p = 0.0029 |
| catecholamine | 3.088 ± 0.087 a | 5.676 ± 0.349 a, c | 10.430 ± 2.156 b, c | ng/μg total protein | YES | F (2, 6) = 8.717 | p = 0.0168 |
| cortisol | 6.024 ± 0.451 a | 11.830 ± 1.165 b | 13.010 ± 1.320 b | pg/μg total protein | YES | F (2, 6) = 12.710 | p = 0.0070 |
| TNF-α | 4.404 ± 0.138 a | 8.020 ± 0.482 a | 12.220 ± 1.465 b | pg/μg total protein | YES | F (2, 6) = 19.150 | p = 0.0025 |
| IL1β | 0.334 ± 0.007 a | 0.631 ± 0.045 a | 1.302 ± 0.117 b | ng/μg total protein | YES | F (2, 6) = 46.850 | p = 0.0002 |
| Hif-1α | 9.727 ± 1.484 a | 22.580 ± 3.800 b | 13.490 ± 2.089 a,b | pg/μg total protein | YES | F (2, 6) = 6.240 | p = 0.0342 |
| creatine kinase | 1.496 ± 0.091 a | 2.820 ± 0.168 b | 2.085 ± 0.243 a,b | pg/μg total protein | YES | F (2, 6) = 13.810 | p = 0.0057 |
| ATP | 142.700 ± 15.940 a | 208.400 ± 20.080 a,b | 112.300 ± 12.880 a,c | ng/μg total protein | YES | F (2, 6) = 8.790 | p = 0.0165 |
| creatinine | 5.035 ± 1.256 | 4.843 ± 0.768 | 5.036 ± 0.850 | U/μg total protein | NO | F (2, 6) = 0.013 | p = 0.9873 |
| metallothionine | 9.871 ± 0.340 | 12.770 ± 1.227 | 11.950 ± 0.820 | pg/μg total protein | NO | F (2, 6) = 2.927 | p = 0.1297 |
Enzymatic or ELISA (enzyme-linked immunosorbent assay)-based methods were applied to detect the ROS (reactive oxygen species), TBARS (thiobarbituric acid reactive substances), SOD (superoxidase dismutase), catalase, ssDNA (single-stranded DNA), catecholamine, cortisol, TNF-α (tumor necrosis factor alpha), IL-1β (interleukin 1 beta), Hif-1α, creatine kinase, ATP (adenosine triphosphate), creatinine, and metallothionein in muscle and gill for C60 NPs-exposed zebrafish, respectively. Data were presented as means ± SEM with three to six independent experiments. Statistical analysis was by one-way ANOVA test followed by a Tukey post-hoc test. Different labels above columns indicate a significant difference between different experimental groups with p < 0.01.