Effects of 25(OH)D₃ and 1.25(OH)₂D₃ on mRNA gene expression of VDR, 1-α hydroxylase and pro-inflammatory markers LPS-induced Mφ.

Mφ (n = 6) were pre-treated with either 25(OH)D3 (100 nM and 500 nM) or 1.25(OH)2D3 (0.05 nM and 10 nM) for 12 hours following LPS challenge (1 μg/mL) for 4 hours. Following targets ((a) VDR, (b) Cyp27b1, (c) TNF-α, (d) MCP-1, (e) NF-κB and (f) IL-6) were analysed by RT-qPCR. Target mRNA gene expression was divided with reference gene GAPDH and ratios were normalised to control Mφ given the value 1. Repeated measures One-way ANOVA Test of trend was performed to evaluate dose-dependent response of either 25(OH)D3 or 1.25(OH)2D3. P-value numbers are stated over the specific groups. One-way ANOVA analysis and Dunnett’s multiple comparisons test was also performed to compare the means of control group with each stimulation group and significance in illustrated as *. (c) * Statistically significant difference between LPS-induced Mφ and Mφ pre-treated with 500 nM 25(OH)D3 and ** with 10 nM 1.25(OH)2D3.