Autophagy is a major cause of pathological vascular remodeling under hypoxic pulmonary hypertension (PH). Sirtuin 3 (Sirt 3) has recently been reported to be involved in the regulation of autophagy, however, its role as an autophagy regulator during hypoxic PH, particularly the molecular mechanism, remains poorly understood. In the present study, Western blot, immunohistochemistry, immunofluorescence, bromodeoxyuridine incorporation and cell cycle analyses were performed to elucidate the underlying mechanism of hypoxia-induced autophagy and cell proliferation with respect to Sirt 3. We observed that the Sirt 3 expression was decreased under hypoxia and that Sirt 3 overexpression significantly inhibited the effects of hypoxia on autophagy. Next, we investigated the mechanistic role of microRNAs in Sirt 3-associated autophagy under hypoxic conditions, with luciferase reporter, microscale thermophoresis and RNA immunoprecipitation assays, results confirming that Sirt 3 is a direct target of miR-874-5p. Furthermore, miR-874-5p was upregulated following hypoxia, and miR-874-5p depletion in turn inhibited autophagy and consequently suppressed abnormal smooth muscle cell proliferation. These findings provide insight into the contribution of the miR-874-5p/Sirt 3 cascade with regard to changes in autophagy and proliferation associated with PH.
Keywords: Autophagy; Hypoxia; Pulmonary hypertension; Sirtuin 3; miR-874-5p.
Copyright © 2020. Published by Elsevier B.V.