Wang Jian, Jin Yingjie, Li Jianchun
Department of Anesthesiology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, P.R. China.
Department of Anesthesiology, Shengze Hospital of Jiangsu, Suzhou, Jiangsu 215228, P.R. China.
Exp Ther Med. 2018 Oct;16(4):3740-3744. doi: 10.3892/etm.2018.6590. Epub 2018 Aug 9.
Neurosurgery always results in neuroinflammation, which may activate microglial cells. Previous studies have demonstrated that fentanyl could be used for the induction or maintenance of anesthesia prior to surgery. However, it is unknown if fentanyl attenuates neuroinflammation prophylactically. Cell viability in groups that were treated with different concentrations of fentanyl (0.01, 0.1, 1 or 5 µmol/l) was analyzed by an MTT assay. BV-2 microglial cells were treated with lipopolysaccharide (LPS) at a concentration of 1 µg/ml to mimic neuroinflammation . BV-2 cells were pretreated with 5 µmol/l fentanyl prior to stimulation by LPS. The protein levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-10 in the culture medium were assessed by ELISA. The mRNA level of toll-like receptor (TLR)4 was evaluated by reverse transcription-quantitative polymerase chain reaction analysis. The protein levels of TLR4, glycogen synthase kinase (GSK)-3β and phosphorylated (p)-GSK-3β in BV-2 cells were assessed by western blot analysis. The MTT assay demonstrated that low concentrations of fentanyl (0.01, 0.1 or 1 µmol/l) did not affect the cell viability of BV-2 cells, while 5 µmol/l fentanyl significantly reduced BV-2 cell viability. The results of ELISA revealed that LPS significantly upregulated the release of TNF-α, IL-1β and IL-10, which were repressed by fentanyl pretreatment. Fentanyl pretreatment significantly reduced the LPS-induced elevation of TLR4 at mRNA and protein levels as well as p-GSK-3β protein levels in BV-2 cells. In conclusion, fentanyl pretreatment protects BV-2 cells from LPS-induced neuroinflammation by inhibiting TLR4 expression and GSK-3β activation. Neuroinflammation induced by surgery serves an important role in the development of postoperative cognitive dysfunction (POCD) and targeting the TLR4 and GSK-3β signaling pathway may provide a novel therapeutic approach for the treatment of POCD.
神经外科手术总会引发神经炎症，这可能会激活小胶质细胞。先前的研究表明，芬太尼可用于手术前麻醉的诱导或维持。然而，芬太尼是否能预防性减轻神经炎症尚不清楚。通过MTT法分析用不同浓度芬太尼（0.01、0.1、1或5μmol/L）处理的组中的细胞活力。用浓度为1μg/ml的脂多糖（LPS）处理BV-2小胶质细胞以模拟神经炎症。在LPS刺激之前，用5μmol/L芬太尼对BV-2细胞进行预处理。通过酶联免疫吸附测定（ELISA）评估培养基中肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β和IL-10的蛋白质水平。通过逆转录-定量聚合酶链反应分析评估Toll样受体（TLR）4的mRNA水平。通过蛋白质印迹分析评估BV-2细胞中TLR4、糖原合酶激酶（GSK）-3β和磷酸化（p）-GSK-3β的蛋白质水平。MTT法表明，低浓度芬太尼（0.01、0.1或1μmol/L）不影响BV-2细胞的细胞活力，而5μmol/L芬太尼显著降低BV-2细胞活力。ELISA结果显示，LPS显著上调TNF-α、IL-1β和IL-10的释放，而芬太尼预处理可抑制这些释放。芬太尼预处理显著降低LPS诱导的BV-2细胞中TLR4在mRNA和蛋白质水平以及p-GSK-3β蛋白质水平的升高。总之，芬太尼预处理通过抑制TLR4表达和GSK-3β激活来保护BV-2细胞免受LPS诱导的神经炎症。手术诱导的神经炎症在术后认知功能障碍（POCD）的发生中起重要作用，针对TLR4和GSK-3β信号通路可能为POCD的治疗提供一种新的治疗方法。
