Regulatory mechanism of ginsenoside Rd on histone H3 acetylation / 中草药

ABSTRACT

Objective Based on quantitative proteomics analysis and molecular biology experimental verification, the regulatory mechanism of ginsenoside Rd on histone H3 acetylation levels was elucidated. Methods The effects of ginsenoside Rd on the dynamic changes of proteome of HEK293T cells were detected by.stable isotope labeling with amino acid (SILAC) technique and LC-MS/MS; Quantitative proteomics database analysis was used to monitor the changes in histone acetyltransferase HATs and histone deacetylase HDACs expression levels. Western blotting and qRT-PCR were used to verify the changes of related protein expression and transcriptional level. Gene knockdown experiments were performed using siRNAs to determine the role of ginsenoside Rd in regulating the level of acetyl modifications at histone H3K9 and K18 sites. Results The histone H3K9ac, H3K18ac expression levels in HEK293T cells decreased after ginsenoside Rd treatment, but the P300 catalytic modification of these two sites did not change significantly; At the same time, ginsenoside Rd up-regulated the transcription and expression of HDAC2, and siHDAC2 treatment reversed the down-regulation effects of ginsenoside Rd on H3K9ac and H3K18ac in HEK293T cells. Conclusion Ginsenoside Rd down-regulates the acetylation level of lysine at histone H3K9 and K18 sites by up-regulating HDAC2, thereby affecting transcriptional activation of downstream genes.