miR-671-5p-enriched exosomes derived from human embryonic stem cells under hypoxia balance oxidative stress homeostasis and macrophage reprogramming to alleviate Legg-Calvé-Perthes disease by targeting NOX2

  • Int Immunopharmacol. 2026 Jun 15:179:116563. doi: 10.1016/j.intimp.2026.116563.
Tao Luo  1 ,  Quan Liu  1 ,  Xiaohua Jiang  2 ,  Tonghua Zhou  1 ,  Shiding Jiang  3 ,  Gangan Liu  3 ,  Xia Lan  4
Affiliations
  • 1. Orthopedic Hospital, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang 330006, Jiangxi, China.
  • 2. Department of General Surgery, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang 330006, Jiangxi, China.
  • 3. Department of Intensive Care Unit, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang 330006, Jiangxi, China.
  • 4. Orthopedic Hospital, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang 330006, Jiangxi, China. Electronic address: [email protected].
PMID: 41931957 DOI: 10.1016/j.intimp.2026.116563
Abstract

Background: Legg-Calvé-Perthes disease (LCPD) remains a pediatric condition that causes hip joint deformities, with complicated pathogenesis. This study explored the influence of hypoxia-preconditioned human embryonic stem cells (hESCs)-derived exosomes on LCPD and its related mechanism.

Methods: Spontaneously hypertensive rats (SHRs) and High mobility group B1 (HMGB1) -stimulated chondrocytes were used to mimic LCPD disease-like lesion in vivo and in vitro. Femoral head damage was examined by hematoxylin-eosin (HE) and Saffron-O and fast green staining. Target molecule expression was determined by real-time quantitative PCR (RT-qPCR), Western blotting, immunohistochemical staining, immunofluorescent staining, and enzyme-linked immunosorbent assay (ELISA). Mitochondrial dysfunction was analyzed by mitochondrial membrane potential and transition pore opening. Cell proliferation was assessed by cell counting kit-8 (CCK-8) and BrdU incorporation. Molecular mechanisms were elucidated by dual luciferase reporter assay, m6A RNA immunoprecipitation (Me-RIP), and total RNA m6A level detection.

Results: Hypoxia preconditioning observably elevated miR-671-5p level in hESCs and their exosomes. Methyltransferase-like 3 (METTL3)-mediated m6A crucially favored miR-671-5p biogenesis by DiGeorge syndrome critical region 8 (DGCR8) in hypoxia-exposed hESCs. Exosomes of hypoxia-exposed hESCs (Exohypoxia) delivered miR-671-5p to significantly attenuate HMGB1-induced oxidative stress injury, mitochondrial dysfunction, and Pyroptosis of chondrocytes via targeting NADPH Oxidase 2 (NOX2). Moreover, miR-671-5p-enriched Exohypoxia restrained macrophage inflammatory reprogramming via specifically inhibiting NOX2-mediated ROS accumulation. Exohypoxia ameliorated LCPD development in rats via modulation of miR-671-5p/NOX2 axis.

Conclusion: Exohypoxia transferred miR-671-5p to alleviate oxidative stress and Pyroptosis of chondrocytes and M1 macrophage reprogramming via targeting NOX2, thereby ameliorating LCPD disease-like lesion.

Keywords
Exosome; Human embryonic stem cells; Legg-Calvé-Perthes disease; Macrophage reprogramming; miR-671-5p.
Products
Inhibitors & Agonists
Other Products