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| Effects of propofol on hypoxia/reoxygenation induced PC12 cell damage and its molecular mechanism |
| XU Jie WANG Hui WEN Hong ZHANG Jian#br# |
Department of Anesthesiology, Beijing Chao-yang Hospital, Capital Medical University, Beijing 100020, China
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Abstract Objective To investigate the effect of propofol on hypoxia/reoxygenation (H/R) induced PC12 cell damage and its molecular mechanism. Methods PC12 cells were cultured in vitro and divided into control group (21% O2/5% CO2/74% N2 culture under normal conditions) and H/R group (3% O2/5% CO2/92% N2 culture under hypoxia condition for 2 h, cultured for 12 h under normal conditions), H/R+ propofol group (50 μmol/L propofol pretreatment for 1 h and then treated with H/R), H/R+anti-miR-NC group (transfected with anti-miR-NC and then treated with H/R), H/R+anti-miR-1246 group (transfected with anti-miR-1246 and then treated with H/R), H/R+ propofol +miR-NC group (transfected miR-NC, and then treated with 50 μmol/L propofol and H/R), H/R+ propofol +miR-1246 (transfected with miR-1246 and then treated with 50 μmol/L propofol and H/R). MTT and flow cytometry were used to detect cell viability and apoptosis, respectively. Western blot analysis of apoptosis-related protein expression (B lymphoblastoma-2 [Bcl-2], B lymphoblastoma-2 associated with x protein [Bax]). The expression levels of pro-inflammatory cytokines and anti-inflammatory cytokines were detected by enzyme-linked immunosorbent assay. The expression level of miR-1246 was detected by quantitative reverse transcription polymerase chain reaction. Results The survival rate of cells was the highest when 50 μmol/L propofol was applied, so the concentration of propofol was 50 μmol/L in subsequent experiments. Compared with control group, the apoptosis rate, levels of Bax protein, tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-18 in H/R group were increased, while the levels of Bcl-2 protein and IL-10 were decreased (P＜0.05). Compared with H/R group, the apoptosis rate, levels of Bax protein, TNF-α, IL-1β, and IL-18 were decreased in H/R+ propofol group, while the levels of Bcl-2 protein and IL-10 were increased (P＜0.05). Compared with control group, the expression of miR-1246 in H/R group was increased (P＜0.05). Compared with H/R group, the expression of miR-1246 in H/R+ propofol group was decreased (P＜0.05). There were no significant differences in miR-1246 level, apoptosis rate, Bcl-2 protein, Bax protein, TNF-α, IL-1β, IL-18, and IL-10 between H/R group and H/R+anti-miR-NC group (P＞0.05). Compared with H/R+anti-miR-NC group, the levels of miR-1246, apoptosis rate, Bax protein, TNF-α, IL-1β, and IL-18 were decreased in H/R+anti-miR-1246 group, while the levels of Bcl-2 protein and IL-10 were increased (P＜0.05). Compared with H/R+ propofol +miR-NC group, the expression level of miR-1246, apoptosis rate, Bax protein, TNF-α, IL-1β, and IL-18 levels were increased in H/R+ propofol +miR-1246 group, while the levels of Bcl-2 protein and IL-10 were decreased (P＜0.05). Conclusion Propofol can relieve H/R induced PC12 cell damage by down-regulating miR-1246.
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