Effects of Different Components of PM2.5 on the Expression Levels of NF-κB Family Gene mRNA and Inflammatory Molecules in Human Macrophage
Abstract
:1. Introduction
2. Materials and Methods
2.1. PM2.5 Sampling and Constituent Preparation
2.1.1. PM2.5 Sampling
2.1.2. PM2.5 Component Extraction and Solution Preparation
2.2. Cell Culture and Exposure to PM2.5
2.3. MTT for the Cell Survival Rate
2.4. ELISA for Inflammatory Molecules and Cytokines in the Supernatant
2.5. Real-Time Quantitative PCR for the Expression Levels of NF-κB Family Genes
2.6. Statistical Analyses
3. Results
3.1. Differentiation of Monocytes
3.2. Effects of PM2.5 on Cell Survival Rate
3.3. Effects of PM2.5 on the Levels of IL-1β, IL-6, IL-8, IL-12 p70, TNF-α, CRP, IL-2, and COX-2
3.4. Effects of PM2.5 on the Relative mRNA Expression Levels of NF-KB Family Gene
3.5. The Connection between the Expression of NF-κB mRNA and Inflammatory Molecules
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Doses | Concentration of PM2.5 (μg/mL) | |||
---|---|---|---|---|
W-PM2.5 | F-PM2.5 | I-PM2.5 | Total PM2.5 | |
Low dose | 75 | 25 | 100 | 200 |
Mid dose | 150 | 50 | 200 | 400 |
High dose | 300 | 100 | 400 | 800 |
Genes | Primer | Sequence |
---|---|---|
β-actin | Forward | 5′-CTGGAACGGTGAAGGTGACA-3′ |
Reverse | 5′-CGGCCACATTGTGAACTTTG-3′ | |
NF-κB1 | Forward | 5′-CACAAGGCAGCAAATAGACGAG-3′ |
Reverse | 5′-TGGGGCATTTTGTTGAGAGTT-3′ | |
NF-κB2 | Forward | 5′-GGCTGGTGCTGACATCCAT-3′ |
Reverse | 5′-CTGCTTCGGGTGTCCTTCTC-3′ | |
RelA | Forward | 5′-CCCCAGCCCTATCCCTTTAC-3′ |
Reverse | 5′-TGCCCAGAAGGAAACACCA-3′ | |
RelB | Forward | 5′-ATGAATGTGGTGAGGATCTGCTT-3′ |
Reverse | 5′-CTCTGATGTGTTTGTGGATTTCTTG-3′ | |
c-Rel | Forward | 5′-GACGACTGCTCTTCCTCCTGTT-3′ |
Reverse | 5′-TCATCTCCTCCTCTGACACTTCC-3′ |
Component | Cytokines | IL-1β | IL-6 | IL-8 | IL-12 p70 | CRP | TNF-α |
---|---|---|---|---|---|---|---|
W-PM2.5 | NF-κB1 | 0.211 | −0.626 | −0.388 | 0.251 | 0.055 | −0.398 |
NF-κB2 | 0.460 | −0.335 | −0.068 | −0.221 | −0.081 | −0.189 | |
RelA | −0.020 | −0.330 | −0.369 | −0.468 | −0.289 | 0.287 | |
RelB | 0.500 | −0.512 | −0.374 | −0.259 | −0.495 | −0.014 | |
c-Rel | 0.479 | −0.519 | −0.278 | −0.085 | −0.305 | −0.030 | |
F-PM2.5 | NF-κB1 | −0.370 | −0.178 | −0.584 | 0.580 | 0.100 | −0.117 |
NF-κB2 | −0.440 | −0.212 | −0.048 | 0.641 * | −0.372 | −0.627 | |
RelA | −0.428 | −0.325 | −0.164 | 0.445 | −0.466 | −0.325 | |
RelB | −0.332 | −0.487 | −0.129 | 0.260 | −0.203 | −0.343 | |
c-Rel | −0.424 | −0.263 | −0.696 * | 0.634 * | 0.047 | −0.156 | |
I-PM2.5 | NF-κB1 | 0.674 * | −0.695 * | 0.605 | −0.173 | −0.131 | −0.424 |
NF-κB2 | 0.636 * | −0.739 * | 0.641 * | −0.256 | −0.043 | −0.356 | |
RelA | 0.681 * | −0.710 * | 0.577 | −0.167 | −0.148 | −0.405 | |
RelB | 0.322 | −0.656 * | 0.212 | −0.093 | −0.175 | −0.444 | |
c-Rel | 0.700 * | −0.681 * | 0.609 | −0.149 | −0.139 | −0.436 |
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Zhu, J.; Zhao, Y.; Gao, Y.; Li, C.; Zhou, L.; Qi, W.; Zhang, Y.; Ye, L. Effects of Different Components of PM2.5 on the Expression Levels of NF-κB Family Gene mRNA and Inflammatory Molecules in Human Macrophage. Int. J. Environ. Res. Public Health 2019, 16, 1408. https://doi.org/10.3390/ijerph16081408
Zhu J, Zhao Y, Gao Y, Li C, Zhou L, Qi W, Zhang Y, Ye L. Effects of Different Components of PM2.5 on the Expression Levels of NF-κB Family Gene mRNA and Inflammatory Molecules in Human Macrophage. International Journal of Environmental Research and Public Health. 2019; 16(8):1408. https://doi.org/10.3390/ijerph16081408
Chicago/Turabian StyleZhu, Jian, Yaming Zhao, Yizhen Gao, Chunyan Li, Liting Zhou, Wen Qi, Yuezhu Zhang, and Lin Ye. 2019. "Effects of Different Components of PM2.5 on the Expression Levels of NF-κB Family Gene mRNA and Inflammatory Molecules in Human Macrophage" International Journal of Environmental Research and Public Health 16, no. 8: 1408. https://doi.org/10.3390/ijerph16081408