Vitamin D Regulates Steroidogenesis and Insulin-Like Growth Factor Binding Protein-1 (IGFBP-1) Production in Human Ovarian Cells. Grishma Parikh 1 , Miroslava Varadinova 1 , Pauline Suwandhi 1 , Takako Araki 1 , Zev Rosenwaks 2 , Leonid Poretsky 1 , Donna Seto-Young 1.
Grishma Parikh1, Miroslava Varadinova1, Pauline Suwandhi1, Takako Araki1, Zev Rosenwaks2, Leonid Poretsky1, Donna Seto-Young1
Mixed ovarian cell culture
Granulosa cell cluture
Mixed Ovarian Cell Culture Factor Binding Protein-1 (IGFBP-1) Production in Human Ovarian Cells
1,25 (OH)2 vitamin D3M199
pM 0 50 100 150
nM 0 50 100 150
Vit D receptor
b-actinRepresentative RT-PCR Analysis of Effects of 1,25-(OH)2D3on Vitamin D Receptor mRNA Expression
Fig. 1.1,25-(OH)2D3 at concentrations of 0-150 pM or 0-150 nM had no effect on expression of vitamin D receptor mRNA whether the cells were incubated in the vitamin D-free MEM Eagle medium (fig. 2) or M199 medium which contains 250 nM vitamin D2.
B Factor Binding Protein-1 (IGFBP-1) Production in Human Ovarian Cells
Granulosa Cell Culture
1,25 (OH)2 vitamin D3MEM M199
pM 0 50 100 150 0 50 100 150
Vit D receptor
Vit D 1-a-
Representative RT-PCR Analysis of Effects of 1,25-(OH)2D3on Vitamin D Receptor and 1-a-Hydroxylase mRNA, 28s rRNA and 18s rRNA Expression
Fi.g. 2. mRNA expression of 1-a-hydroxylase, the enzyme that converts 25-(OH) D to its active form 1,25-(OH)2D, was absent in the granulosa cell cultures
The amount of mRNA used for each set of conditions was similar when examined by mRNA expression of house protein b-actin or 28s rRNA or 18s rRNA
Fig 3. 1,25-(OH)2D3 increased progesterone production in mixed ovarian cell culture by 13% compared to control, but had no significant effect on testosterone production (A)
1,25-(OH)2D3 stimulated estradiol or estrone production in purified granulosa cell culture by up to 6% and up to 21% compared to control, respectively (B).
Fig. 4. In the granulosa cell culture, insulin stimulated estradiol production by 13% (p<0.04). 1,25-(OH)2D3 acted synergistically with insulin to increase estradiol production by up to 50% (p<0.001)
In contrast, insulin and 1,25-(OH)2D3 had no significant synergistic effect on progesterone, testosterone or estrone production (data not shown).
Fig. 5.1,25-(OH)2D3 increased IGFBP-1 production by up to 24% compared to control (p<0.001) in purified granulosa cells.
Fig. 6. In the presence of insulin, 1,25-(OH)2D3, enhanced insulin-induced inhibition of IGFBP-1 production by 13% (p<0.001).