Autophagy induced by hypoxia in pulpitis is mediated by HIF-1α/BNIP3.

Objective: Hypoxia-inducible factor-1α (HIF-1α) and its downstream factor, 19 kDa BCL-2 interacting protein 3 (BNIP3), promote cellular autophagy under hypoxic conditions. However, their roles in pulpitis are unclear. Therefore, the changes in inflammatory response and autophagy levels caused by hypoxia during pulpitis were evaluated. Additionally, the regulatory mechanism of HIF-1α/BNIP3 in cellular autophagy in pulpitis was explored.

Design: Pulp from dental pulp tissues of healthy individuals and patients with pulpitis (n = 10) were exposed and combined with a low oxygen simulation chamber to construct pulpitis (n = 6), hypoxia (n = 6), and hypoxia+pulpitis (n = 6) rat models. Hematoxylin and eosin and immunohistochemical staining were used to detect the localization and expression levels of HIF-1α, BNIP3, and autophagy marker protein, LC3B. Transmission electron microscopy was used to confirm autophagosome formation. An in vitro hypoxic model of human dental pulp cells was established, and HIF-1α chemical inhibitor 3-(5'-hydroxymethyl-2'-furyl)- 1-benzylindazole (YC-1) was administered. Immunofluorescence and western blotting were used to detect the localization and protein levels of HIF-1α, BNIP3, and LC3B.

Results: Autophagy is significantly increased and HIF-1α and BNIP3 are elevated in inflamed dental pulp tissue. Both pulp exposure and hypoxia intervention cause inflammatory reactions in rat dental pulp tissue, accompanied by the autophagy activation. Hypoxia significantly enhances HIF-1α/BNIP3 and autophagy activation. BNIP3 downregulates and autophagy reduces after treatment with YC-1.

Conclusions: In pulpitis, activation of the HIF-1α/BNIP3 signaling pathway driven by hypoxia leads to increased autophagy. This provides a new molecular explanation for autophagy activation in apical periodontitis and new insights into the pathogenesis of the disease.