Research Paper Volume 12, Issue 17 pp 17235—17256

Mitophagy impairment is involved in sevoflurane-induced cognitive dysfunction in aged rats

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Figure 5. Rapamycin relieved sevoflurane-induced mitochondria impairment. H4 cells were exposed to 0% sevoflurane with rapamycin (1μmol/L) (Rapa), 4.1% sevoflurane without rapamycin (SEV), 4.1% sevoflurane with rapamycin (SEV+Rapa), or 0% sevoflurane without rapamycin (Ctrl) for 6 h, and the ROS level (A) and intracellular ROS and mitochondrial ROS levels (B) were measured. Eighteen-month-old rats were subjected to 2% sevoflurane (SEV and SEV+Rapa groups) for 5 h. Rapamycin (20 mg/kg/d) was administrated intraperitoneally two days before sevoflurane treatment, and daily administrations were continued for one week (Rapa and SEV+Rapa groups). After perfusion, the ultrastructure of mitochondria in the hippocampus was observed under an electron microscope. (C) The lengths of all mitochondria were measured. (G) The Tomm20 and LAMP1 protein levels were measured by immunofluorescence assay. Scale bar represents 50 μm. (D) The results of Tomm20 (E) and LAMP1 (F) quantification are shown. The Manders’ overlap coefficient is shown in (H). Images show representative examples from three independent experiments for each group. The data are expressed as mean ± SD. *P<0.05, Ctrl vs SEV, Rapa and SEV+Rapa; #P<0.05, SEV vs Rapa and SEV+Rapa; ^P<0.05, Rapa vs SEV+Rapa.