(W1130-04-26) Peonidin-3-O-Glucoside and Resveratrol Reduce Human hFOB Osteoblast Apoptosis and Alter the Expression of Genes Associated with Apoptosis, Differentiation and Osteoclastogenesis

Purpose: Data from systems biology approaches, including mRNA-seq and miRNA-seq have shown that nutrients and combinations of nutrients play an important role in bone homeostasis and remodeling by impacting numerous signaling pathways. Previously, we have reported that the dietary compounds resveratrol (RES) and peonidin-3-O-glucoside (POG) reduced osteoblast apoptosis and increased differentiation, and reduced osteoclastogenesis in cultured hFOB human osteoblasts and transgenic medaka. In this work, we use whole-genome transcriptomic profiling in POG and RES treated osteoblasts to provide a more comprehensive understanding of the changes in gene expression and associated canonical pathways impacted.

Methods: Cultured human hFOB osteoblasts were treated with RES or POG or vehicle solvent, harvested and total RNA was isolated and purified. RNA-seq libraries (Universal Plus mRNASeq kit) were prepared, and sequencing was carried using a NovaSeq 6000. General quality-control metrics were acquired using FastQC, and STAR. Quantifications of ENSEMBL genes was performed using FeatureCounts, and EdgeR and DSecq2 were used to compute differential expression statistics. Specific gene expression was validated using qPCR.

Results: Transcriptomic analysis showed that of 29,762 genes analyzed, in osteoblast treated with POG, the expression of 3177 genes was significantly altered (1481 up-regulated and 1696 down-regulated, FDR<0.05), while in the RES treated RES osteoblasts, the expression of 2288 genes were significantly altered (1068 up-regulated and 1220 down-regulated, FDR<0.05). IngenuityÒ Pathway analysis of differentially expressed genes revealed that POG altered gene expression in the apoptosis, RANKL signaling and the hypoxia inducible factor pathways.

Conclusion: Peonidin-3-O-glucoside and resveratrol reduce human hFOB osteoblast apoptosis and alter the expression of genes associated with apoptosis, differentiation and osteoclastogenesis.