Joint Toxicity of a Multi-Heavy Metal Mixture and Chemoprevention in Sprague Dawley Rats
<p>Effects of exposure to a heavy metal mixture on spatial learning and memory function, with or without the addition of EGCG, TCD, or GSH. The Morris water test was performed immediately after six months of treatment. Bars represented the times of crossing platform; (<b>A</b>) escape latency results of SD rats; (<b>B</b>) platform crossing times of SD rats; <span class="html-italic">n</span> = 6 rats/each group; Results were presented as means ± SD; Two-way ANOVA analysis was used in (<b>A</b>); One-way ANOVA analysis with LSD-t test was performed in (<b>B</b>). * <span class="html-italic">p</span> < 0.05, vs. the control group (Mixture, Mix + EGCG, Mix + TCD, Mix + GSH compared with the control group); <sup>#</sup> <span class="html-italic">p</span> < 0.05, vs. the Mixture group (Mix + EGCG, Mix + TCD, Mix + GSH compared with the Mixture group); Mix, Mixture.</p> "> Figure 2
<p>Effects of exposure to a heavy metal mixture on histomorphology of liver and testicle with or without the addition of EGCG, TCD or GSH. The formalin-fixed tissues were stored at 4 °C and the histopathological examination lasted two months. Mix, Mixture.</p> "> Figure 3
<p>Contents of eight heavy metals in the heart and liver. ICP-MS was carried out after histopathological examination. Bars represented contents of eight heavy metals (ug) in per gram of organs; (<b>A</b>) Contents of eight heavy metals in the heart of SD rats; (<b>B</b>) Contents of eight heavy metals in the liver of SD rats; <span class="html-italic">n</span> = 6 rats/each group; Results were presented as means ± SD; One-way ANOVA and LSD-t test were showed; * <span class="html-italic">p</span> < 0.05, vs. the control group (Mixture, Mix + EGCG, Mix + TCD, Mix + GSH compared with the control group); <sup>#</sup> <span class="html-italic">p</span> < 0.05, vs. the Mixture group (Mix + EGCG, Mix + TCD, Mix + GSH compared with the Mixture group); Mix, Mixture.</p> "> Figure 4
<p>Contents of eight heavy metals in the spleen and lung of SD rats. Bars represented contents of eight heavy metals (ug) in per gram of organs; (<b>A</b>) Contents of eight heavy metals in the spleen of SD rats; (<b>B</b>) Contents of eight heavy metals in the lung of SD rats; <span class="html-italic">n</span> = 6 rats/each group; Results were presented as means ± SD; One-way ANOVA and LSD-t test were performed; * <span class="html-italic">p</span> < 0.05, vs. the control group (Mixture, Mix + EGCG, Mix + TCD, Mix + GSH compared with the control group); <sup>#</sup> <span class="html-italic">p</span> < 0.05, vs. the Mixture group (Mix + EGCG, Mix + TCD, Mix+GSH compared with the Mixture group); Mix, Mixture.</p> "> Figure 5
<p>Contents of eight heavy metals in the kidney and brain of SD rats. Bars represented contents of eight heavy metals (ug) in per gram of organs; (<b>A</b>) Contents of eight heavy metals in the kidney of SD rats; (<b>B</b>) Contents of eight heavy metals in the brain of SD rats; <span class="html-italic">n</span> = 6 rats/each group; Results were presented as means ± SD; One-way ANOVA and LSD-t test were used. * <span class="html-italic">p</span> < 0.05, vs. the control group (Mixture, Mix + EGCG, Mix + TCD, Mix + GSH compared with the control group); <sup>#</sup> <span class="html-italic">p</span> < 0.05, vs. the Mixture group (Mix + EGCG, Mix + TCD, Mix + GSH compared with the Mixture group); Mix, Mixture.</p> "> Figure 6
<p>Contents of eight heavy metals in the testicle of SD rats. Bars represented contents of eight heavy metals (ug) in per gram of organs; <span class="html-italic">n</span> = 6 rats/each group; Results were presented as means ± SD; One-way ANOVA and LSD-t test were used. * <span class="html-italic">p</span> < 0.05, vs. the control group (Mixture, Mix + EGCG, Mix + TCD, Mix + GSH compared with the control group); Mix, Mixture.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Methods
2.2.1. Animal Experiment
2.2.2. Preparation of Multi-Heavy Metal Mixture
2.2.3. Organ Weight and Organ/Body Weight Coefficients
2.2.4. Hematological and Biochemical Analysis
2.2.5. Histopathological Examination
2.2.6. Determination of Heavy Metals in Serum and Organs by ICP-MS
2.3. Statistics
3. Results
3.1. Morris Water Maze Test
3.2. Organ Weight and Organ/Body Weight Coefficients
3.3. Hematological and Biochemical Analysis
3.4. Histopathological Examination
3.5. ICP-MS Results
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Chemical | Content (mg) | Mass Ratio of Metal Element |
---|---|---|
K2Cr2O7 | 8.547 | 41.345 |
MnCl2•4H2O | 45.609 | 173.264 |
NiCl2•6H2O | 8.893 | 30.054 |
CuSO4•5H2O | 89.816 | 312.824 |
ZnSO4•7H2O | 344.187 | 1070.838 |
CdCl2•2.5H2O | 1.972 | 13.281 |
CH3ClHg | 0.091 | 1.000 |
(CH3COO)2Pb3H2O | 0.884 | 6.611 |
Total | 500.001 |
(g) | Group | F | p | ||||
---|---|---|---|---|---|---|---|
Control | Mixture | Mix + EGCG | Mix + TCD | Mix + GSH | |||
Heart | 1.68 ± 0.11 | 1.66 ± 0.12 | 1.54 ± 0.27 | 1.70 ± 0.13 | 1.72 ± 0.24 | 0.984 | 0.430 |
Liver | 12.20 ± 0.98 | 12.58 ± 1.69 | 12.41 ± 1.48 | 12.88 ± 1.66 | 13.39 ± 1.82 | 0.647 | 0.633 |
Spleen | 0.86 ± 0.12 | 0.77 ± 0.13 | 0.75 ± 0.11 | 0.81 ± 0.13 | 0.89 ± 0.07 | 1.938 | 0.128 |
Lung | 1.67 ± 0.27 | 2.21 ± 0.52 | 1.75 ± 0.26 | 1.83 ± 0.48 | 1.77 ± 0.41 | 1.958 | 0.125 |
Kidney | 3.33 ± 0.33 | 3.28 ± 0.29 | 3.27 ± 0.16 | 3.51 ± 0.28 | 3.52 ± 0.31 | 1.531 | 0.217 |
Brain | 2.11 ± 0.10 | 2.09 ± 0.11 | 2.06 ± 0.05 | 2.09 ± 0.18 | 2.10 ± 0.08 | 0.136 | 0.968 |
Testicle | 3.39 ± 0.17 | 3.16 ± 0.15 * | 2.96 ± 0.21 * | 3.18 ± 0.21 * | 3.16 ± 0.19 * | 3.923 | 0.011 |
Group | F | p | |||||
---|---|---|---|---|---|---|---|
Control | Mixture | Mix + EGCG | Mix + TCD | Mix + GSH | |||
Heart | 0.32 ± 0.02 | 0.32 ± 0.03 | 0.29 ± 0.05 | 0.30 ± 0.02 | 0.30 ± 0.04 | 0.983 | 0.431 |
Liver | 2.29 ± 0.18 | 2.39 ± 0.38 | 2.31 ± 0.17 | 2.28 ± 0.14 | 2.29 ± 0.14 | 0.327 | 0.858 |
Spleen | 0.16 ± 0.02 | 0.15 ± 0.02 | 0.14 ± 0.03 | 0.14 ± 0.02 | 0.15 ± 0.02 | 1.167 | 0.344 |
Lung | 0.31 ± 0.04 | 0.42 ± 0.10 * | 0.32 ± 0.05 | 0.33 ± 0.08 | 0.30 ± 0.07 | 3.025 | 0.032 |
Kidney | 0.63 ± 0.09 | 0.62 ± 0.06 | 0.61 ± 0.03 | 0.63 ± 0.04 | 0.60 ± 0.03 | 0.295 | 0.879 |
Brain | 0.39 ± 0.02 | 0.40 ± 0.03 | 0.39 ± 0.03 | 0.37 ± 0.04 | 0.36 ± 0.03 | 1.752 | 0.163 |
Testicle | 0.64 ± 0.02 | 0.60 ± 0.03 | 0.55 ± 0.07 * | 0.57 ± 0.05 * | 0.54 ± 0.06 * | 3.949 | 0.011 |
Parameters | Group | F | p | ||||
---|---|---|---|---|---|---|---|
Control | Mixture | Mix + EGCG | Mix + TCD | Mix + GSH | |||
White Blood Cell | |||||||
WBC(×109/L) | 4.47 ± 0.93 | 3.27 ± 0.57 | 3.63 ± 0.91 | 3.20 ± 0.82 | 2.23 ± 0.47 | 3.362 | 0.055 |
NE% | 31.57 ± 10.74 | 35.20 ± 9.10 | 27.17 ± 1.96 | 29.67 ± 2.60 | 30.40 ± 3.22 | 0.592 | 0.676 |
LY% | 65.73 ± 9.90 | 61.46 ± 8.63 | 69.57 ± 3.41 | 65.73 ± 3.14 | 63.10 ± 4.72 | 1.176 | 0.353 |
MO% | 1.07 ± 1.16 | 3.53 ± 1.57 | 1.43 ± 1.25 | 2.43 ± 1.16 | 3.03 ± 0.15 | 2.421 | 0.117 |
EO% | 1.63 ± 0.35 | 2.80 ± 2.21 | 1.83 ± 0.49 | 1.60 ± 0.26 | 1.40 ± 0.61 | 0.800 | 0.552 |
Red Blood Cell | |||||||
RBC(×1012/L) | 7.08 ± 0.35 | 7.16 ± 0.50 | 7.22 ± 0.37 | 7.12 ± 0.26 | 7.15 ± 0.40 | 0.093 | 0.984 |
HGB(g/L) | 138.00 ± 6.32 | 137.17 ± 6.40 | 137.60 ± 5.50 | 136.83 ± 5.52 | 128.50 ± 17.07 | 1.069 | 0.393 |
HCT | 0.40 ± 0.02 | 0.42 ± 0.03 | 0.41 ± 0.02 | 0.41 ± 0.03 | 0.39 ± 0.06 | 0.411 | 0.799 |
MCV(fl) | 56.67 ± 2.45 | 57.50 ± 1.54 | 56.30 ± 1.27 | 57.28 ± 1.63 | 57.60 ± 1.58 | 0.417 | 0.795 |
MCH(pg) | 19.78 ± 0.66 | 19.15 ± 0.55 | 19.10 ± 0.37 | 19.48 ± 0.67 | 19.40 ± 0.44 | 1.399 | 0.264 |
MCHC(g/L) | 343.33 ± 8.16 | 333.17 ± 8.26 | 339.40 ± 6.11 | 340.00 ± 12.33 | 337.17 ± 9.24 | 1.011 | 0.421 |
RDW-CV(%) | 14.62 ± 1.09 | 15.58 ± 0.97 * | 14.68 ± 0.64 # | 14.73 ± 0.69 | 13.97 ± 0.38 # | 3.107 | 0.034 |
Platelet | |||||||
PLT(×109/L) | 881.33 ± 139.71 | 823.83 ± 73.52 | 777.60 ± 73.43 | 833.17 ± 40.09 | 849.67 ± 51.80 | 1.441 | 0.251 |
PDW-CV(%) | 14.67 ± 0.05 | 14.65 ± 0.05 | 14.70 ± 0.10 | 14.70 ± 0.06 | 14.67 ± 0.05 | 0.675 | 0.616 |
MPV(fl) | 5.78 ± 0.08 | 6.00 ± 0.14 * | 5.94 ± 0.17 * | 6.05 ± 0.08 * | 5.97 ± 0.12 * | 4.195 | 0.010 |
PCT | 0.51 ± 0.08 | 0.53 ± 0.04 | 0.46 ± 0.04 | 0.52 ± 0.04 | 0.51 ± 0.03 | 1.487 | 0.237 |
Parameters | Group | F | p | ||||
---|---|---|---|---|---|---|---|
Control | Mixture | Mix + EGCG | Mix + TCD | Mix + GSH | |||
TG(mmol/L) | 0.62 ± 0.07 | 0.76 ± 0.45 | 0.77 ± 0.19 | 0.68 ± 0.39 | 0.68 ± 0.15 | 0.265 | 0.897 |
CHOL(mmol/L) | 1.29 ± 0.30 | 1.39 ± 0.56 | 1.37 ± 0.28 | 1.29 ± 0.39 | 1.14 ± 0.18 | 0.417 | 0.794 |
TBIL(μmol/L) | 2.85 ± 0.57 | 3.73 ± 0.84 * | 2.37 ± 0.50 # | 2.72 ± 0.54 # | 2.50 ± 0.39 # | 3.667 | 0.021 |
DBIL(μmol/L) | 0.83 ± 0.39 | 1.17 ± 0.47 | 0.57 ± 0.06 | 0.90 ± 0.31 | 0.75 ± 0.26 | 1.911 | 0.148 |
IBIL(μmol/L) | 1.90 ± 0.36 | 2.57 ± 0.59 * | 1.80 ± 0.56 # | 1.82 ± 0.44 # | 1.75 ± 0.18 # | 3.337 | 0.031 |
TP(g/L) | 53.80 ± 2.80 | 56.17 ± 3.88 | 57.48 ± 2.62 | 53.98 ± 3.29 | 54.07 ± 3.00 | 1.469 | 0.243 |
ALB(g/L) | 27.30 ± 1.17 | 29.03 ± 1.00 | 29.44 ± 1.22 | 27.90 ± 2.03 | 28.17 ± 1.78 | 1.620 | 0.202 |
GLOB(g/L) | 26.50 ± 1.38 | 27.13 ± 2.94 | 28.04 ± 1.46 | 26.08 ± 1.46 | 25.90 ± 1.41 | 1.208 | 0.333 |
A/G(ALB/GLOB) | 1.03 ± 0.05 | 1.08 ± 0.08 | 1.05 ± 0.03 | 1.07 ± 0.05 | 1.09 ± 0.04 | 1.033 | 0.411 |
ALT(U/L) | 41.50 ± 7.74 | 45.67 ± 4.27 | 51.20 ± 6.67 | 42.17 ± 4.26 | 45.33 ± 9.50 | 1.705 | 0.182 |
AST(U/L) | 145.83 ± 35.10 | 104.50 ± 30.46 | 146.80 ± 49.16 | 139.83 ± 17.17 | 129.33 ± 27.50 | 1.673 | 0.189 |
ALP(U/L) | 83.17 ± 13.66 | 129.67 ± 22.49 * | 119.40 ± 23.33 * | 100.33 ± 8.98 # | 96.67 ± 22.18 # | 5.710 | 0.002 |
UREA(mmol/L) | 6.74 ± 1.11 | 6.35 ± 0.62 | 6.80 ± 0.62 | 5.90 ± 0.82 | 6.17 ± 0.66 | 1.310 | 0.295 |
CREA(μmol/L) | 35.07 ± 3.58 | 34.32 ± 3.17 | 33.64 ± 2.32 | 31.33 ± 1.13 | 32.08 ± 3.69 | 1.642 | 0.196 |
UR/Cr | 0.19 ± 0.02 | 0.19 ± 0.02 | 0.20 ± 0.02 | 0.19 ± 0.03 | 0.20 ± 0.03 | 0.495 | 0.739 |
URCA(μmol/L) | 81.83 ± 12.19 | 83.50 ± 17.75 | 90.60 ± 13.28 | 77.00 ± 15.77 | 78.00 ± 18.24 | 0.636 | 0.642 |
Element (µg/L) | Group | F | p | ||||
---|---|---|---|---|---|---|---|
Control | Mixture | Mix + EGCG | Mix + TCD | Mix + GSH | |||
52Cr | 7.164 ± 0.624 | 9.271 ± 1.171 * | 7.503 ± 0.607 | 8.197 ± 1.127 | 8.464 ± 1.441 * | 3.129 | 0.035 |
55Mn | 5.829 ± 0.974 | 9.123 ± 4.492 | 6.695 ± 1.409 | 6.374 ± 1.673 | 7.671 ± 3.327 | 1.267 | 0.311 |
60Ni | 0.150 ± 0.245 | 3.012 ± 1.496 * | 0.230 ± 0.124 # | 0.305 ± 0.264 # | 0.426 ± 0.294 # | 11.251 | <0.001 |
63Cu | 1056.750 ± 104.982 | 950.482 ± 107.573 | 964.510 ± 116.376 | 1010.820 ± 110.527 | 1122.620 ± 103.817 | 1.034 | 0.411 |
66Zn | 1092.200 ± 125.269 | 1062.810 ± 125.269 | 1104.850 ± 100.431 | 1126.280 ± 162.298 | 1126.970 ± 137.700 | 0.252 | 0.905 |
111Cd | 0.112 ± 0.096 | 0.826 ± 1.277 | 0.320 ± 0.191 | 0.253 ± 0.117 | 0.308 ± 0.119 | 1.198 | 0.338 |
202Hg | 0.000 ± 0.000 | 9.712 ± 1.059 * | 8.847 ± 1.356 * | 10.476 ± 1.315 * | 10.350 ± 0.968 * | 111.310 | <0.001 |
208Pb | 1.422 ± 0.402 | 2.164 ± 1.106 | 1.710 ± 0.824 | 1.911 ± 0.496 | 2.126 ± 1.056 | 0.813 | 0.530 |
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Wang, Y.; Tang, Y.; Li, Z.; Hua, Q.; Wang, L.; Song, X.; Zou, B.; Ding, M.; Zhao, J.; Tang, C. Joint Toxicity of a Multi-Heavy Metal Mixture and Chemoprevention in Sprague Dawley Rats. Int. J. Environ. Res. Public Health 2020, 17, 1451. https://doi.org/10.3390/ijerph17041451
Wang Y, Tang Y, Li Z, Hua Q, Wang L, Song X, Zou B, Ding M, Zhao J, Tang C. Joint Toxicity of a Multi-Heavy Metal Mixture and Chemoprevention in Sprague Dawley Rats. International Journal of Environmental Research and Public Health. 2020; 17(4):1451. https://doi.org/10.3390/ijerph17041451
Chicago/Turabian StyleWang, Yafei, Yuqing Tang, Zhou Li, Qihang Hua, Li Wang, Xin Song, Baobo Zou, Min Ding, Jinshun Zhao, and Chunlan Tang. 2020. "Joint Toxicity of a Multi-Heavy Metal Mixture and Chemoprevention in Sprague Dawley Rats" International Journal of Environmental Research and Public Health 17, no. 4: 1451. https://doi.org/10.3390/ijerph17041451