Effects of Vitamin D analogs on gene expression profiling in human coronary artery smooth muscle cells J. Ruth Wu-Wong, Masaki Nakane, Junli Ma, Xiaoan.

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Effects of Vitamin D analogs on gene expression profiling in human coronary artery smooth muscle cells J. Ruth Wu-Wong, Masaki Nakane, Junli Ma, Xiaoan Ruan, Paul E. Kroeger Atherosclerosis Volume 186, Issue 1, Pages 20-28 (May 2006) DOI: 10.1016/j.atherosclerosis.2005.06.046 Copyright © 2005 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 The effects of paricalcitol and calcitriol: the genes regulated by either paricalcitol or calcitriol are shown in hierarchical clustering. Atherosclerosis 2006 186, 20-28DOI: (10.1016/j.atherosclerosis.2005.06.046) Copyright © 2005 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 Effects of paricalcitol on IGF1, WT1 and TGFβ3 mRNA expression. Smooth muscle cells were harvested, RNA isolated and the mRNA levels of target genes analyzed by real-time RT-PCR. GADPH was used for normalization. Statistical analysis was performed by unpaired t-test. *p<0.05, **p<0.01, ***p<0.001 compared with control; n=3 per condition. Atherosclerosis 2006 186, 20-28DOI: (10.1016/j.atherosclerosis.2005.06.046) Copyright © 2005 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 Effects of paricalcitol and calcitriol on THBS1, BNP and CYP24A1 mRNA expression. Smooth muscle cells were harvested, RNA isolated and the mRNA levels of target genes analyzed by real-time RT-PCR. GADPH was used for normalization. Statistical analysis was performed by unpaired t-test. *p<0.05, **p<0.01, ***p<0.001 compared with control; n=3 per condition. Atherosclerosis 2006 186, 20-28DOI: (10.1016/j.atherosclerosis.2005.06.046) Copyright © 2005 Elsevier Ireland Ltd Terms and Conditions

Fig. 4 Effect of paricalcitol and calcitriol on cell proliferation. Cells were treated with increasing concentrations of paricalcitol or calcitriol as described in Section 2. Data were expressed as percentage of control (cells in growth medium without drug treatment, 100%). Each value shown is mean±the standard deviation (n=4–8 from combining two independent experiments). Serum free medium was used as a comparison in the studies (21.4±5.3% of control). Statistical comparisons were performed by unpaired t-test. **p<0.01, ***p<0.001 compared with control. Atherosclerosis 2006 186, 20-28DOI: (10.1016/j.atherosclerosis.2005.06.046) Copyright © 2005 Elsevier Ireland Ltd Terms and Conditions

Fig. 5 Effects of Vitamin D analogs in the blood vessel. VDR activation by Vitamin D analogs results in regulation of genes involved in the cell cycle that leads to inhibition of proliferation and induction of differentiation. BNP (natriuretic peptide-B) is down-regulated. Regulation of genes, such as TM, thrombospondin 1 and MMP by Vitamin D analogs results in reduced thrombogenicity and increased fibrinolysis. The regulation of type-B endothelin receptor, oxytocin receptor and prostaglandin-endoperoxide synthase 1 suggest that Vitamin D analogs may also play roles in vessel relaxation and endothelial regeneration. Atherosclerosis 2006 186, 20-28DOI: (10.1016/j.atherosclerosis.2005.06.046) Copyright © 2005 Elsevier Ireland Ltd Terms and Conditions