Effects of Betulinic Acid on the Proliferation, Cellular Senescence, and Type 1 Interferon-Related Signaling Pathways...

Pentacyclic triterpenoids, including betulinic acid (BA), and their glycosides are abundant in fruits such as Zizyphus sp., Dillenia sp., and Azanza sp. These compounds exhibit various pharmacological activities in human cells. Here, we investigated the effects of BA on the cellular proliferation and senescence of cultured normal human dermal fibroblasts (NHDFs). BA treatment for 24-48 h increased the proliferation of low-passage young fibroblasts. Furthermore, BA reduced the proportion of senescent cells, as determined via the β-galactosidase assay of high-passage NHDFs. DNA microarray analysis and subsequent validations via quantitative real-time polymerase chain reaction revealed that BA downregulates interferon (IFN)-inducible genes, including IFIT1, IFITM1, IFI6, MX1, and OAS2, which are upregulated in replicative senescent cells compared with the low-passage young cells (control). Enrichment analysis based on the microarray data predicted BA-induced suppression of the type I IFN signaling pathway. BA downregulated the expression of the IRF9 transcriptional factor downstream of the type 1 IFN signaling pathway. IFN-inducible genes were downregulated via IRF9 silencing using siRNA compared with the negative control treated with siRNA. Consistently, BA treatment reduced the proportion of senescent cells and IFN-inducible genes in etoposide-treated fibroblasts. Hence, BA alleviates cellular senescence via the inhibition of the type 1 IFN signaling pathway in dermal fibroblasts.