# Preparation of Artemisia argyi-Derived Extracellular Nanovesicles and Their Protective Effects Against Oxidative Stress-Induced Senescence in Endometrial Stromal Cells

## TL;DR

This study shows that nanovesicles from Artemisia argyi protect endometrial cells from oxidative stress by reducing ROS and restoring mitochondrial function.

## Contribution

The study introduces Artemisia argyi-derived nanovesicles as a natural reservoir of antioxidants that protect against oxidative stress via the SIRT1/PGC-1α/Nrf2 pathway.

## Key findings

- A-NVs reduce ROS levels and enhance SOD activity in H2O2-damaged endometrial stromal cells.
- A-NVs activate the SIRT1/PGC-1α/Nrf2 pathway, reducing senescence markers like SA-β-Gal and p21.
- Blocking SIRT1 with EX-527 inhibits the protective effects of A-NVs, confirming the pathway's role.

## Abstract

Oxidative stress-induced endometrial injury has been shown to contribute to infertility; however, effective strategies that can simultaneously scavenge reactive oxygen species (ROS) and restore mitochondrial and antioxidant homeostasis remain elusive. In this study, we isolated extracellular nanovesicles from Artemisia argyi (A-NVs) and investigated their protective effects on H2O2-damaged human endometrial stromal cells (hESCs). We discovered that A-NVs possess a typical lipid bilayer structure and contain a variety of bioactive components. Our metabolomic analysis indicates that A-NVs can be regarded as a “natural drug reservoir”, in which flavonoids account for approximately 10.8%. We demonstrate that A-NVs can be efficiently taken up by cells, improve cell viability, reduce intracellular and mitochondrial ROS levels, enhance superoxide dismutase (SOD) activity, upregulate the expression of catalase (CAT), SOD1, and SOD2, and partially restore mitochondrial membrane potential. Mechanistically, A-NVs exert antioxidant effects by activating the SIRT1/PGC-1α/Nrf2 signaling axis. SIRT1 activation further alleviates H2O2-induced premature senescence, as evidenced by a 71.8% reduction in SA-β-Gal-positive cells compared with the H2O2 group, together with downregulation of p53 and p21 expression. These positive protective effects can be blocked by the SIRT1 inhibitor EX-527, confirming the central role of this pathway. Collectively, our findings demonstrate that A-NVs can maintain redox and mitochondrial homeostasis while inhibiting oxidative stress-related senescence progression, underscoring their application potential in endometrial repair and functional recovery.