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Objective: To observe the Asian sodium arsenite, DMA (price), MMA (price) of human skin fibroblasts proliferation to explore the molecular mechanisms of arsenic skin lesions. Method: 1. Primary cultured skin fibroblasts for the study, MTT (MTT) reduction method for detection of different concentrations of sodium arsenite, DMA (5 price), MMA (price) (0 umol / 1,0.5 μmol / 1,1 μmol/15μmol/1, 10μmol / 1,15 μmol / 1) in human skin fibroblasts proliferation. Flow cytometry different concentrations of sodium arsenite, sodium arsenite, DMA (5 price), MMA (price) (0μmol / L, 0.5 micromol / l, 5 micromol / L, 10 micromol / l) treatment skin fibroblasts cell cycle distribution. 3 TEM observation of different concentrations of sodium arsenite into the fiber cell ultrastructure of the skin. 4 semi-quantitative RT-RT-PCR to detect different concentrations of sodium arsenite, sodium arsenate, DMA (5 price), MMA (price) treatment of human skin into fibroblasts after DNMT1, DNMT3A, DNA Methyltransferase 3B in AS3MT mRNA expression. Results: 1.MTT reduction test results show that: Compared with the control group, the human skin fibroblast cells were treated with different concentrations of sodium arsenite, DMA (5 price), after the induction of the MMA (price), the final concentration of 0.5 micromol / l, 1 micromol / l, 5 micromol / l sodium arsenite, DMA (price), MMA (price) absorbance values ??were increased in a dose - response relationship (P lt; 0.05), a final concentration of 10 micromol / l 15 micromol / l of sodium arsenite, DMA (price), MMA (price) absorbance values ??were lower, there is a dose - response relationship (P lt; 0.05); primary cultured skin fibroblasts were observed under fluorescence microscope to the 13th generation of slow growth; normal skin fibroblast growth curve can be observed 96 hours after the skin into fibroblast growth faster trend. 2 TEM observation 0.5 micromol / l sodium arsenite role in skin fibroblasts, cell surface microvilli, thin and long, large nuclei, irregular core, prominent nucleoli, a small number of organelles in the cytoplasm, the cell mitochondrial vacuolation; 10Lmol / l sodium arsenite role in skin fibroblast cell surface microvilli than 5 micromol / l concentration was significantly lysosomes compared 5Lmol / 1 concentrations increased, the expansion of the endoplasmic reticulum, and no nuclear Ren, mitochondrial vacuolation. 3 flow cytometry cell cycle distribution results show that low doses of sodium arsenite, sodium arsenate, DMA (price), MMA (price) treated cells in G1 phase cells relative to the control group, the percentage reduction in does not appear G1 arrest; high dose of sodium arsenite, sodium arsenate, DMA (5 price), MMA (price) treated cells in G1 phase the percentage increase in cell cycle arrest in G1 phase. DNMT3A, DNMT1 mRNA expression of the DNA Methyltransferase 3B in as sodium arsenite, MMA drug effect dose increased the expression of a downward trend, and with the increase in the dose of sodium arsenite, DMA drug effects, mRNA expression levels increasing trend the mRNA of AS3MT, in addition to low doses of sodium arsenite treated DNMT1 mRNA expression compared with control group was not statistically significant, other trends were statistically significant and mRNA expression (P lt; 0.05) than the control group; expression levels with sodium arsenite, MMA drug effect dose increases the expression of an increasing trend, and with sodium arsenite, DMA drug effects increasing doses of a downward trend in the amount of mRNA expression. In addition to the low dose of sodium arsenite, high doses of sodium arsenite on the skin into fibroblasts cells AS3MT mRNA expression compared with control group difference was not statistically significant (P gt; 0.05), other trends have the system to learn significance (P lt; 0.05). Conclusion: The low concentrations of sodium arsenite, DMA (5 price), MMA (price) to stimulate human skin fibroblast proliferation, high concentrations of cytotoxic;, in doses of sodium arsenite, sodium arsenate, DMA ( pentavalent), MMA (price) so that the skin fibroblast cell cycle arrest in G1 phase, low dose does not appear G1 arrest; electron microscopy showed mitochondrial vacuolation with the the sodium arsenite drug concentration increase in skin fibroblasts cell surface microvilli gradually reduced nucleolar edge set → nucleolar components separation → nucleolus; different concentrations of sodium arsenite, sodium arsenate, DMA (5 price), MMA (price) affect DNA methylation transferase gene (DNMT3A, DNMT1 DNMT3B, As3MT) expression, which is important in the study of the molecular mechanisms of arsenic skin lesions.
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