Abstract:

Objective To explore the mechanism of action of lncRNA H19（H19）/Sirt3 mediated autoph⁃ agy in injury after intracerebral hemorrhage（ICH）. Methods Amouse model of ICH was established by injections with collagenase. SH⁃SY5Y cells treated with oxyhemoglobin（OxyHb）were used as an in vitro ICH model. Alenti⁃ viral vector containing sh⁃H19 was used to inhibit H19 expressions in ICH mice and cultured SH⁃SY5Y cells. 48 hours after establishment of the ICH model，the mouse neurologic deficits were analyzed by mNSS score and lesion volume. RT⁃qPCR，Tunel staining and immunofluorescence were used to analyze the expression of H19，neuronal apoptosis and LC3 express in the injured tissues respectively. Cell viability was measured by MTT assay，apoptosis was detected by Tunel staining，and expressions of autophagy ⁃ related proteins and Sirt3/AMPK/mTOR signaling pathway were analyzed by Western blot. Results The expression of H19 was increased after ICH and in OxyHb⁃ treated cells in a time⁃dependent manner（P < 0.05）. H19 knockdown reduced the lesion volume and mNSS score on the second day after establishment of the ICH model（P < 0.01），decreased neuronal apoptosis（P < 0.01）， and enhanced regulation of autophagy during ICH injury. H19 knockdown enhanced the viability of SH⁃SY5Y cells exposed to OxyHb，reduced apoptosis，and accelerated autophagy. H19 accelerated autophagy mainly through regu⁃ lating Sirt3 expression and activating AMPK⁃mTOR pathway. Conclusions H19 is enhanced in a time⁃dependent manner in ICH models in vivo and in vitro. H19 knockdown relieves ICH damage via autophagy induced by Sirt3/ AMPK/mTOR signaling pathway.

Key words:

lncRNA H19, Sirt3, AMPK/mTOR signaling pathway, intracerebral hemorrhage, au? tophagy, mouse