目的：探讨丙泊酚通过沉默信息调节因子1（silent information regulator 1,SIRT1）/核转录因子E2相关因子2（nuclear factor-erythroid 2-related factor 2,Nrf2）途径调控脑缺血/再灌注（I/R）损伤大鼠模型中铁死亡的作用机制。方法：构建大鼠脑I/R模型和体外使用氧糖剥夺/复氧（OGD/R）处理小鼠海马神经元细胞HT22,并用丙泊酚预先给药处理。通过TTC染色分析大鼠脑梗死体积。Western blot检测大鼠脑组织中SIRT1/Nrf2信号和铁死亡相关蛋白（GPX-4和Ptgs2）表达,试剂盒检测大鼠脑组织中活性氧（ROS）,丙二醛（MDA）,超氧化物歧化酶（SOD）和Fe2+水平。分别在大鼠体内和HT22细胞中干扰SIRT1,以及在HT22细胞中同时干扰SIRT1和过表达Nrf2,观察大鼠脑组织中SIRT1/Nrf2信号相关蛋白以及铁死亡变化。HT22细胞中添加自噬抑制剂3-MA处理后,检测自噬相关蛋白表达和铁死亡变化。结果：I/R组大鼠脑梗死体积高于Sham组,丙泊酚组低于I/R组。I/R组大鼠脑组织中SIRT1、Nrf2、HO-1、NQO1蛋白表达高于Sham组,丙泊酚组低于I/R组。较Sham组,I/R组大鼠脑组织中ROS,MDA,Fe2+水平增加,SOD含量减少,Ptgs2表达增加,GPX-4表达减少,丙泊酚逆转了这些结果。与丙泊酚组相比,干扰SIRT1增加I/R大鼠脑组织和ODG/R后HT22细胞中ROS,MDA和Fe2+水平,减少SOD水平,过表达Nrf2逆转了这些结果。ODG/R后HT22细胞中LC3Ⅱ/Ⅰ比例上调,丙泊酚下调了这些结果,干扰SIRT1上调了LC3Ⅱ/Ⅰ比例,自噬抑制剂处理下调了这些结果,降低了ROS,MDA和Fe2+水平,增加SOD水平（P<0.05）。结论：丙泊酚通过激活SIRT1/Nrf2途径抑制自噬减少铁死亡进而保护大鼠脑I/R损伤。
Abstract
Objective To explore the mechanism of propofol in regulating ferroptosis through SIRT1/Nrf2 pathway in rats with cerebral ischemia/reperfusion (I/R) injury. Methods The rat brain I/R model was constructed and the mouse hippocampal neuron HT22 cells were treated with oxygen and glucose deprivation/reoxygenation (OGD/R) in vitro and pre-treated with propofol. Cerebral infarct volume was analyzed by TTC staining. Western blot was carried out to determine related protein levels of SIRT1/Nrf2 signaling and ferroptosis in the brain tissue of rats. Reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD) and Fe2+ levels in the brain tissue of rats were detected by corresponding kits. SIRT1 was silenced in rats and HT22 cells, and SIRT1 and Nrf2 were silenced simultaneously in HT22 cells. The changes of SIRT1/Nrf2 signaling related proteins and ferroptosis in rat brain tissue were detected. HT22 cells were treated with autophagy inhibitor 3-MA, and the expression of autophagy-related proteins and ferroptosis changes were detected. Results The cerebral infarct volume of the I/R group was higher than those of the Sham group, and the propofol group were lower than those of the I/R group. The protein expressions of SIRT1, Nrf2, HO-1 and NQO1 in brain tissue of rats in I/R group were higher than those in Sham group, and those in propofol group were lower than I/R group. Compared with Sham group, ROS, MDA, Fe2+ and Ptgs2 levels in the brain tissue of rats in I/R group were increased, and SOD and GPX-4 levels were decreased, and propofol reversed these results (P<0.01). Compared with the propofol group, SIRT1 silencing increased ROS, MDA and Fe2+ levels and decreased SOD level in the brain tissue of I/R rats and HT22 cells after ODG/R, and overexpression of Nrf2 reversed these results. After ODG/R, the ratio of LC3Ⅱ/Ⅰ were increased in HT22 cells, and propofol down-regulated these results. The silence of SIRT1 up-regulated the ratio of LC3Ⅱ/Ⅰ, and the autophagy inhibitor treatment down-regulated these results, reduced the levels of ROS, MDA and Fe2+, and increased the level of SOD. Conclusion Propofol inhibits autophagy and reduces ferroptosis through activation of SIRT1/Nrf2 pathway to protect brain I/R injury in rats.
关键词
丙泊酚 /
SIRT1/Nrf2 /
脑缺血/再灌注（I/R）损伤 /
自噬 /
铁死亡
Key words
propofol /
SIRT1/Nrf2 /
cerebral ischemia/reperfusion (I/R) injury /
autophagy /
ferroptosis
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基金
江苏大学临床医学科技发展基金（JLY20180214）
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