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- Reins, Rose Y., et al.
(författare)
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Vitamin D Induces Global Gene Transcription in Human Corneal Epithelial Cells : Implications for Corneal Inflammation
- 2016
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Ingår i: Investigative Ophthalmology and Visual Science. - : Association for Research in Vision and Ophthalmology (ARVO). - 0146-0404 .- 1552-5783. ; 57:6, s. 2689-2698
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Tidskriftsartikel (refereegranskat)abstract
- PURPOSE. Our previous studies show that human corneal epithelial cells (HCEC) have a functional vitamin D receptor (VDR) and respond to vitamin D by dampening TLR-induced inflammation. Here, we further examined the timing of the cytokine response to combined vitamin D-TLR treatment and used genome-wide microarray analysis to examine the effect of vitamin D on corneal gene expression. METHODS. Telomerase-immortalized HCEC (hTCEpi) were stimulated with polyinosinicpolycytidylic acid (poly[I: C]) and 1,25-dihydroxyvitamin D-3 (1,25D(3)) for 2 to 24 hours and interleukin (IL)-8 expression was examined by quantitative (q) PCR and ELISA. Telomeraseimmortalized HCEC and SV40-HCEC were treated with 1,25D(3) and used in genome-wide microarray analysis. Expression of target genes was validated using qPCR in both cell lines and primary HCEC. For confirmation of IjBa protein, hTCEpi were treated with 1,25D(3) for 24 hours and cell lysates used in an ELISA. RESULTS. Treatment with 1,25D(3) increased poly(I: C)-induced IL-8 mRNA and protein expression after 2 to 6 hours. However, when cells were pretreated with 1,25D(3) for 24 hours, 1,25D(3) decreased cytokine expression. For microarray analysis, 308 genes were differentially expressed by 1,25D(3) treatment in hTCEpi, and 69 genes in SV40s. Quantitative (q) PCR confirmed the vitamin D-mediated upregulation of target genes, including nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (I kappa B alpha). In addition to increased transcript levels, I kappa B alpha protein was increased by 28% following 24 hours of vitamin D treatment. CONCLUSIONS. Microarray analysis demonstrates that vitamin D regulates numerous genes in HCEC and influences TLR signaling through upregulation of I kappa B alpha. These findings are important in dissecting the role of vitamin D at the ocular surface and highlight the need for further research into the functions of vitamin D and its influence on corneal gene expression.
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| 2. |
- Thrikawala, Savini, et al.
(författare)
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Triazole fungicides induce adipogenesis and repress osteoblastogenesis in zebrafish
- 2023
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Ingår i: Toxicological Sciences. - : Oxford University Press (OUP). - 1096-6080 .- 1096-0929. ; 193:2, s. 119-130
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Tidskriftsartikel (refereegranskat)abstract
- Triazoles are a major group of azole fungicides commonly used in agriculture, and veterinary and human medicine. Maternal exposure to certain triazole antifungal medication causes congenital malformations, including skeletal malformations. We hypothesized that triazoles used as pesticides in agriculture also pose a risk of causing skeletal malformations in developing embryos. In this study, teratogenic effects of three commonly used triazoles, cyproconazole, paclobutrazol, and triadimenol, were investigated in zebrafish, Danio rerio. Exposure to the triazole fungicides caused bone and cartilage malformations in developing zebrafish larvae. Data from whole-embryo transcriptomics with cyproconazole suggested that exposure to this compound induces adipogenesis while repressing skeletal development. Confirming this finding, the expression of selected bone and cartilage marker genes were significantly downregulated with triazoles exposure as determined by quantitative PCR. The expression of selected adipogenic genes was upregulated by the triazoles. Furthermore, exposure to each of the three triazoles induced adipogenesis and lipid droplet formation in vitro in 3T3-L1 pre-adipocyte cells. In vivo in zebrafish larvae, cyproconazole exposure caused lipid accumulation. These results suggest that exposure to triazoles promotes adipogenesis at the expense of skeletal development, and thus they expand the chemical group of bona fide bone to fat switchers.
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