摘要
目的探讨七氟醚对缺氧诱导的小胶质细胞炎症反应的影响,及对转录因子NF-E2相关因子2(NRF2)及血红素加氧酶1(HO-1)/高迁移率族蛋白B1(HMGB1)通路的调控作用。方法缺氧不同时间(0、2、4、6、8、12 h)诱导BV-2小胶质细胞活化建立炎症模型,倒置显微镜观察细胞活化形态并计算活化细胞比例;Western blot法检测小胶质细胞NRF2、HO-1、HMGB1及白介素1β(IL-1β)蛋白表达水平,筛选最佳缺氧时间8 h。观察缺氧8 h条件下七氟醚不同浓度(0%、2%、4%、6%)处理30 min对NRF2、HO-1、HMGB1、IL-1β蛋白表达的影响。将BV-2细胞分为正常对照组、缺氧诱导组、6%七氟醚组、SnPP组(NRF2/HO-1通路抑制剂-锡原卟啉)、6%七氟醚+SnPP组,计算活化细胞比例并检测细胞NRF2/HO-1/HMGB1通路蛋白及核转录因子-κB p65(NF-κB p65)、磷酸化NF-κB p65(p-NF-κB p65)、IL-1β蛋白表达水平。结果随着缺氧时间延长,活化细胞比例及HMGB1、IL-1β表达逐渐升高,NRF2、HO-1蛋白在8~12 h时下降至最低水平(P<0.05),实验选用8 h为缺氧诱导条件。随着七氟醚浓度升高,活化细胞比例、HMGB1、IL-1β表达逐渐减少,NRF2、HO-1逐渐升高(P<0.05),且呈剂量依赖性,选取6%七氟醚为处理浓度。与正常对照组比较,缺氧诱导组NRF2、HO-1表达降低,活化细胞比例、HMGB1、IL-1β、p-NF-κB p65/NF-κB p65表达升高(P<0.05)。6%七氟醚组上述指标变化与缺氧诱导组相反(P<0.05);SnPP组上述指标变化与缺氧诱导组一致且比缺氧诱导组严重(P<0.05)。6%七氟醚+SnPP组上述指标变化与6%七氟醚组相反(P<0.05)。结论七氟醚可通过促进NRF2/HO-1活化,抑制HMGB1表达,来减轻缺氧诱导的MG活化及炎症反应进程。
Objective To investigate the effects of sevoflurane on hypoxia-induced inflammatory responses of microglia and regulation of the nuclear factor erythroid 2-related factor 2(NRF2)/heme oxygenase 1(HO-1)/high mobility group protein B1(HMGB1)pathway.Methods BV-2 mouse microglia were activated by inducing hypoxia for 0,2,4,6,8 and 12 h to establish an inflammation model.The activated cells were observed by inverted microscope,and the proportion of activated cells was calculated. Western blot were used to detect the expression levels of NRF2, HO-1, HMGB1 and interleukin (IL)-1β proteins in microglia. The optimal hypoxia time was 8 h. BV-2 microglia were activated under hypoxic conditions for 8 h;treated with different concentrations of sevoflurane (0%, 2%, 4% and 6%) for 30 min;and NRF2, HO-1, HMGB1, IL-1β protein expression was observed. BV-2 cells were divided into a control group, hypoxia-induction group, 6% sevoflurane group, SnPP group ( NRF2/ HO-1 pathway inhibitor stannous porphyrin), and 6% sevoflurane + SnPP group. The ratio of activated cells was calculated, and the protein expression levels of NRF2, HO-1, HMGB1, nuclear factor-κB p65 (NF-κB p65), phosphorylated NF-κB p65 (p-NF-κB p65) and IL-1β were determined. Results With prolonged hypoxia time, the proportion of activated cells and expression of HMGB1 and IL-1β gradually increased, and NRF2 and HO-1 proteins decreased to the lowest level at 8 ~ 12 h (P<0. 05). Therefore, the hypoxia-induction time of 8 h was chosen. With the increase in sevoflurane concentration, the proportion of activated cells and expression of HMGB1 and IL-1β gradually decreased, and NRF2 and HO-1 expression gradually increased (P<0. 05). These effects were dose-dependent, and 6% sevoflurane was selected as the treatment concentration. Compared with that in the control group, the expression of NRF2 and HO-1 in the hypoxia-induced group decreased, and the proportion of activated cells and expression of HMGB1, IL-1β and p-NF-κB p65/ NF-κB p65 expression increased (P <0. 05). The changes in the above indices in the 6% sevoflurane group were opposite to those in the hypoxia-induced group (P<0. 05). The changes in the SnPP group were consistent with those in hypoxia-induced group and were more serious than those in hypoxia-induced group (P<0. 05). The changes in the above parameters in the 6% sevoflurane + SnPP group were opposite to those in 6% sevoflurane group (P <0. 05). Conclusions Sevoflurane treatment reduced hypoxia-induced microglial activation and inflammatory processes by promoting NRF2/ HO-1 activation and inhibiting HMGB1 expression.
作者
温翔宇
张军宏
朱存良
WEN Xiangyu;ZHANG Junhong;ZHU Cunliang(Department of Anesthesiology,Dingxi People’s Hospital of Gansu Province,Dingxi 743000,China;Department of Urology,Dingxi People’s Hospital of Gansu Province,Dingxi 743000;Loudi Central Hospital of Hunan Province,Loudi 417000)
出处
《中国比较医学杂志》
CAS
北大核心
2022年第2期66-73,共8页
Chinese Journal of Comparative Medicine
基金
2020年度娄底市科技创新项目(娄科发〔2020〕27号)。
作者简介
温翔宇(1983—),女,硕士,主治医师,研究方向:临床麻醉。E-mail:wxyzhjh83@163.com。
