Nutraceuticals Effects On Your Body's Stem Cells

Figure 1:

          In Figure 1, results of gene expression analysis are shown following the exposure of MSCs to varying nutraceuticals. Figure 1 is a bar graph representation illustrating PCR used to measure target genes known to be subject to epigenetic regulation by both HDAC inhibitors. The graph shows the result of human MSCs treated with varying nutraceuticals alone. The expression of Oct 3/4, a well-known pluripotency gene, was increased about 20-fold compared to untreated human MSCs. The expression of SIRT-1 was highly elevated compared to untreated MSC by ~300-fold without differences between treatment with either VPA or Curcumin. The expression of FGF-21 was also elevated by 25-fold and its expression was higher in MSCs treated with VPA or Curcumin, although this increase is not significant.  CXCR4 expression was highest in Curcumin treated MSCs, which is a well-known marker of stem cell migration activity. The expression of Hsp70 was elevated by 25-fold and its expression was highest in MSCs treated with VPA and Curcumin, but not significant. All gene expression was normalized to untreated MSCs. Beta actin was measured as a house-keeping gene. The graph shows the result of gene expression analysis of human MSCs treated with varying nutraceuticals compared to VPA. Gene expression was quantified by determining the amount of gene-specific DNA/total cDNA. Data is mean +/- SD of 4 replicates. These results thus show highest increased expression of Oct 3/4, SIRT-1, FGF-21, CXCR4, and Hsp70 as a result of exposure to VPA and Curcumin.

Nutraceuticals Effect On Your Body's Stem Cells: Figure 2

          Figure 2 is a line graph representation illustrating migration of CB-MSCs induced by exposure to 500nM curcumin and querectin (black sqaures), curcumin and querectin inhibited by CXCR4 inhibitor (AMD3100) (red circles) and curcumin and querectin inhibited by MMP9 inhibitor (GM6001) (blue triangles). Percent closure is plotted as a function of dose and the data was modeled by sigmoidal curve fitting to calculate EC₅₀ values. This Figure 2 shows curcumin and querectin-induced MSC migration with a calculated EC₅₀ of 316.56 nM and maximal migration of the combination of curcumin and querectin. The CXCR4 inhibitors showed inhibition of CB-MSC with a closure of 2%. The MMP9 inhibitors also showed inhibition of CB-MSC with a closure of 2%. The results showed a lower EC₅₀ than that observed with VPA only, i.e., 32.96 μM suggesting a synergistic effect of CC-Quer on CB-MSC migration. 

The Effect of Nutraceuticals on Your Body's Stem Cells: Figure 3

          Figure 3 is a line graph representation illustrating migration of CB-MSCs induced by exposure to 500nM curcumin and resveratrol (black sqaures), nM curcumin and resveratrol inhibited by CXCR4 inhibitor (AMD3100) (red circles) and nM curcumin and resveratrol inhibited by MMP9 inhibitor (GM6001) (blue triangles). Percent closure is plotted as a function of dose and the data was modeled by sigmoidal curve fitting to calculate EC₅₀ values. This Figure 3 shows curcumin and resveratrol-induced MSC migration with a calculated EC₅₀ of 253.19 nM and maximal migration of the combination of curcumin and resveratrol. The CXCR4 inhibitors showed inhibition of CB-MSC with a closure of 1%. The MMP9 inhibitors also showed inhibition of CB-MSC with a closure of 1%. The results showed a lower EC₅₀ than that observed with VPA only, i.e., 32.96 μM suggesting a synergistic effect of CC-Res on CB-MSC migration. 

Nutraceuticals As A Replacement To Pharmaceuticals 

          The molecular mechanisms of the effect of lithium and VPA were then investigated on stem cell migration. To understand the effects of nutraceuticals as a replacement to pharmaceuticals, curcumin, quercetin and resveratrol were investigated to compare lithium and VPA results. Since a prior report suggested that VPA up-regulated CXCR4, a critical chemokine receptor involved with cellular mobility, and that lithium up-regulated MMP-9 (Tsai, LK, et al., Stroke 42(10): 2932-2939, 2011), the effects of known inhibitors of CXCR4 and MMP-9 on the migration of CB-MSCs were determined and the results are shown in Figure 2 and 3, which is a line graph representation illustrating migration of human MSCs exposed to a combination of nutraceutics with and without inhibition of MMP9 and CXCR4.  Percent closure is plotted as a function of dose. The results indicate that the CXCR-4 inhibitor, AMD 3100, blocked the curcumin-induced CB-MSC migration and that GM 6001, a competitive inhibitor of MMP-9, quercetin induced CB-MSCs, and resveratrol induced CB-MSCs. These results suggest that CXCR4 and MMP-9 are molecular components of the curcumin and quercetin-induced migration, and curcumin and resveratrol-induced migration of CB-MSCs thus confirming the replacement of these pharmaceuticals with these nutraceuticals.