Cadmium–Zinc Interaction in Mus musculus Fibroblasts
Abstract
:1. Introduction
2. Results
2.1. Cell Viability
2.2. Accumulation of Cd
2.3. Accumulation of Zn
2.4. GSH Content
2.5. Metallothionein Content
2.6. MT-1 mRNA Gene Expression
2.7. MT-2 mRNA Gene Expression
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Treatment with Heavy Metals
4.3. Determination of Metals and MT Contents
4.4. Glutathione Quantification
4.5. qRT-PCR Analysis
4.6. Statistical Analysis
5. Conclusions
- In co-treatments, both Cd and Zn are accumulated in less quantity and this determines the greater tolerance to Cd-induced cytotoxicity.
- In the WT strain in the treatment with Zn alone, there does not appear to be an increase in GSH content, which probably occurs only when cells are subjected to oxidative stress. In contrast, treatments with Cd alone cause a reduction in GSH due to the probable oxidative stress that this metal entails. Instead, in co-treatments, Zn causes an increase in the GSH content compared to treatments with Cd alone, promoting its expression and preventing oxidation through indirect mechanisms, demonstrating its protective role.
- The MTF-1 null strain, although lacking the MTF-1 factor, still shows a high GSH content, generally higher than that of the WT strain, thanks to an alternative mechanism of expression of y-glutamylcysteine synthetase, independent of MTF-1.
- The two strains have different threshold doses of activation of MT production: lower for the MTF-1 null strain and higher for the WT strain.
- The MTF-1 null strain expresses only isoform 1 of the MT through the USF/ARE sequences, making up for the lack of isoform 2 and ensuring a higher protein production than that of the WT strain for treatments with Cd only.
- The time courses of the mRNAs of the MT-1 and MT-2 genes and those of the protein found are variable, caused by the different half-lives of the messenger and protein and/or by the intervention of down-regulation mechanisms of gene expression.
- Although the MTF-1 null strain in the literature is considered more sensitive than the WT strain, it generally exhibits greater viability following Cd-only treatments than the WT strain, possibly due to the more significant amount of MT it presents.
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
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4 h | 18 h | 24 h | ||
---|---|---|---|---|
µg Cd/mg protein | ||||
WT | Cd 1 | 0.002 ± 0.001 a* | 0.033 ± 0.003 a# | 0.029 ± 0.005 a# |
Cd 2 | 0.004 ± 0.0001 a* | 0.006 ± 0.0018 b# | 0.168 ± 0.0077 b° | |
ZnCd 1 | 0.027 ± 0.0013 b* | 0.018 ± 0.0002 c# | 0.021 ± 0.0002 a° | |
ZnCd 2 | 0.029 ± 0.0023 b* | 0.031 ± 0.0003 d* | 0.038 ± 0.0008 c# | |
MTF-1 null | Cd 1 | 0.021 ± 0.0016 a* | 0.090 ± 0.0059 a# | 0.022 ± 0.001 a* |
Cd 2 | 0.030 ± 0.0003 b* | 0.266 ± 0.0479 b# | 0.088 ± 0.0141 b* | |
ZnCd 1 | 0.013 ± 0.0001 c* | 0.023 ± 0.0005 c# | 0.055 ± 0.0012 c° | |
ZnCd 2 | 0.014 ± 0.0001 c* | 0.018 ± 0.0008 c# | 0.051 ± 0.0017 c° | |
µg Zn/mg protein | ||||
WT | C | 0.027 ± 0.0049 a* | 0.024 ± 0.0119 a* | 0.022 ± 0.0022 a* |
Zn | 0.105 ± 0.0053 b* | 0.050 ± 0.001 b# | 0.066 ± 0.0038 b° | |
Cd 1 | 0.023 ± 0.0008 a* | 0.017 ± 0.0002 a# | 0.023 ± 0.0038 a° | |
Cd 2 | 0.025 ± 0.0003 a* | 0.016 ± 0.0004 a# | 0.065 ± 0.003 b° | |
ZnCd 1 | 0.030 ± 0.0014 a* | 0.040 ± 0.0004 b# | 0.054 ± 0.0006 c° | |
ZnCd 2 | 0.039 ± 0.0031 c* | 0.040 ± 0.0004 b* | 0.057 ± 0.0011 c° | |
MTF-1 null | C | 0.024 ± 0.0104 a* | 0.024 ± 0.0064 a* | 0.028 ± 0.0015 a* |
Zn | 0.044 ± 0.0005 b* | 0.099 ± 0.0046 b# | 0.080 ± 0.0038 b° | |
Cd 1 | 0.026 ± 0.002 a* | 0.027 ± 0.0018 a* | 0.024 ± 0.001 a* | |
Cd 2 | 0.023 ± 0.0002 a* | 0.067 ± 0.012 c# | 0.040 ± 0.0063 c° | |
ZnCd 1 | 0.031 ± 0.0003 a* | 0.039 ± 0.0008 d# | 0.089 ± 0.002 d° | |
ZnCd 2 | 0.032 ± 0.0002 a* | 0.041 ± 0.0017 d# | 0.063 ± 0.0021 e° |
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Priante, E.; Pietropoli, E.; Piva, E.; Santovito, G.; Schumann, S.; Irato, P. Cadmium–Zinc Interaction in Mus musculus Fibroblasts. Int. J. Mol. Sci. 2022, 23, 12001. https://doi.org/10.3390/ijms231912001
Priante E, Pietropoli E, Piva E, Santovito G, Schumann S, Irato P. Cadmium–Zinc Interaction in Mus musculus Fibroblasts. International Journal of Molecular Sciences. 2022; 23(19):12001. https://doi.org/10.3390/ijms231912001
Chicago/Turabian StylePriante, Ettore, Edoardo Pietropoli, Elisabetta Piva, Gianfranco Santovito, Sophia Schumann, and Paola Irato. 2022. "Cadmium–Zinc Interaction in Mus musculus Fibroblasts" International Journal of Molecular Sciences 23, no. 19: 12001. https://doi.org/10.3390/ijms231912001