Fig. 1

Graphic illustration of the experimental procedure. a In this section, the mice underwent behavioral tests on days 1, 3, and 7 after long-term isoflurane exposure. Hippocampus was immediately harvested at the end of the behavioral testing, and ELISA analysis for IL-1β and IL-6 levels was performed. The changes of Cx43 configuration were measured by immunofluorescence and Western blot. b The primary astrocytes were harvested 6 h after isoflurane exposure, and immunofluorescence and Western blot were performed to detect alterations of the Cx43 configuration. ELISA was performed to measure the level of IL-1β and IL-6 expression. Scrape-loading dye transfer was used to evaluate the biological function of Cx43 gap junction channels. c Schematic diagram of in vitro Cx43 gap junctions rescue experiment. Improvement of astrocyte pathological changes induced by long-term isoflurane anesthesia was observed by administration of ZP1609, a specific enhancer of the Cx43 gap junction. d Schematic diagram of in vivo Cx43 gap junctions functional rescued experiments. For GJs-Cx43-mediated astrocytic network intervention, mice were treated with ZP1609 (dosage: 300 µg/kg every time, three times/day) by intraperitoneal injection for 3 consecutive days after the mice model was established. Behavioral tests were conducted on day 3 after anesthesia. ELISA was performed to confirm the expression levels of IL-1β and IL-6. The changes of Cx43 configuration were measured by immunofluorescence and Western blot. Dye transfer was used to evaluate the biological function of Cx43 gap junction channels. Ethidium bromide (EtBr) uptake assays were used to measure the activity of Cx43 hemichannels. Transmission electron microscopy was used to observe ultrastructural alterations of astrocyte gap junction in mice hippocampal tissue