To explore the anti-aging effects of chlorogenic acid (CGA) and the underlying mechanisms based on a Caenorhabditis elegans (C. elegans) model.

The anti-aging activity of CGA was studied based on the body length, exercise behavior, lipofuscin content, antioxidative stress ability, swallowing frequency, body-bending frequency, and head-swinging ability of C. elegans. Through DAF-16 nuclear translocation and SOD-3-GFP fluorescence experiments, the effects of CGA on ROS levels, antioxidant enzyme activities, MDA content, mutant-strain lifespan, and anti-aging molecular signaling pathways were explored, as well as the underlying mechanisms.

CGA improved multiple indices of the nematode: body length was increased (all P < .001), head-swing frequency and body-bending frequency were increased (all P < .05), nematode longevity was prolonged (P = .0021), lipofuscin deposition in nematodes was slowed down (all P < .001), the chemotaxis index was improved (P = .0012), ROS levels were reduced (all P < .001), and SOD activity and MDA content were reduced (SOD: P = .0017 between the low-concentration group and the control group, P < .001 between the high-concentration and medium-concentration groups and the control group; MDA: P = .0135 between the low-concentration group and the control group, and P < .001 between the high-concentration and medium-concentration groups and the control group). In addition, CGA also activated the DAF-16 transcription factor, promoted DAF-16 nuclear translocation under oxidative stress conditions (both P < .001 between the high-concentration and medium-concentration groups and the control group), and increased SOD-3 gene expression in nematodes (all P < .001).

CGA plays an anti-aging role in C. elegans. The underlying mechanisms include activation of the insulin/IGF-1 signaling pathway and enhancement of DAF-16 activity. This study lays a foundation for further research into the anti-aging effects of CGA.

N-Alkylamides (NAAs), derived from Anacyclus pyrethrum (L.) DC, have potential anti-tumor effects. To explore the molecular mechanism and chemo-preventive capacity of NAAs, we synthesized an NAA (H-10) and evaluated whether it could inhibit the proliferation of B16, HepG2, HeLa, and HCT116 cancer cells in 2D culture.

To evaluate the antiproliferative activity of H-10 in 2D and 3D culture of BD, HepG2, HeLa, and HCT116 cells, multicellular tumor spheroids were constructed to more accurately reflect the cell tumor environment. To visualize nuclear changes related to apoptosis, Hoechst 33258 staining and propidium iodide-Annexin V double staining were performed.

Compound H-10 strongly inhibited the growth of all tested cell lines. Hoechst 33258 staining and propidium iodide-Annexin V double staining revealed that H-10 did not cause morphological alterations in the nuclei and moderately induced late apoptosis only when treated at 180 μM. The strongest inhibitory effect was observed in HCT116 cells. Flow cytometry analysis indicated that treatment of HCT116 cells with compound H-10 resulted in robust cell growth arrest in G2 phase in 2D and 3D cell culture; in 3D-cultured HCT116 cells, growth arrest occurred in G1 phase. Treatment with compound H-10 also significantly suppressed angiogenesis of chick chorioallantoic membrane in vivo.

Treatment with compound H-10 strongly affected clonogenic survival (in the long-term) and migration of HCT116 cells. Therefore, H-10, a compound of NAAs may be useful for treating cancer because of its anti-neoplastic effect and easy synthesis.