芬戈莫德(FTY720)通过激活Nrf2/HO-1通路促进大鼠脑缺血再灌注损伤后神经功能恢复

FTY720 promotes recovery of nerve function after cerebral ischemia reperfusion injury in rats by activating Nrf2/HO-1 pathway

目的研究芬戈莫德(FTY720/fingolimod)对脑缺血再灌注损伤的治疗作用并探索其保护机制。方法60只SD大鼠随机分为假手术组、脑缺血再灌注损伤(MCAO/R)模型组及FTY720组。MCAO/R组和FTY720组用Longa缝合法建立MCAO/R模型。采用改良神经功能损伤评分法评估大鼠神经功能;HE染色观察大脑皮层(额颞叶)神经细胞形态;实时定量PCR及Western blot法分别检测核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)、还原型烟酰胺腺嘌呤二核苷酸磷酸醌氧化还原酶1(NQO-1)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6的mRNA及蛋白水平;化学试剂盒检测超氧化物岐化酶(SOD)活性、丙二醛(MDA)含量。结果与MCAO/R组相比,FTY720组大鼠神经损伤评分降低,细胞破坏减轻,Nrf2、HO-1及NQO-1上调,TNF-α、IL-6及IL-1β下调,SOD活性提高,MDA含量减少。结论FTY720促进MCAO/R后大鼠神经功能恢复,可能与激活Nrf2/HO-1信号通路有关。

Objective To investigate the protective effect of fingolimod(FTY720)on cerebral ischemia/reperfusion injury and explore whether it is related to the inhibition of the nuclear factor E2-related factor 2/heme-oxygenase 1(Nrf2/HO-1)signaling pathway.Methods Sprague-Dawley(SD)rats were equally divided into 3 groups:sham group,cerebral ischemia/reperfusion(MCAO/R)group and FTY720 group.MCAO/R models were established by Longa suture method in the latter two groups.The modified neurological severity scores were used to evaluate neurological function and HE staining to observe morphological features of SD rat nerve cells in the cerebral cortex.In the case of 0.5 mg/kg FTY720,the mRNA and protein levels of nuclear factor E2 related factor 2(Nrf2),heme oxygenase 1(HO-1),quinone oxidoreductase-1(NQO-1),and tumor necrosis factorα(TNF-α),interleukin 1β(IL-1β),Interleukin 6(IL-6)were detected by real-time quantitative PCR and Western blot analysis.Simultaneously,the antioxidant enzyme activity(superoxide dismutase,SOD)and the content of lipid peroxidation products(malondialdehyde,MDA)were tested by chemical kits.Results Compared with the MCAO/R group,the FTY720 group showed the increased number of cells,effectively improved morphology and lower nerve damage scores.In the MCAO/R group,the expression of Nrf2,HO-1 and NQO-1 significantly decreased and the expression of TNF-α,IL-6 and IL-1 increased markedly,while this situation was reversed obviously by FTY720.At the same time,the improvement of antioxidant capacity was reflected in the increase of SOD activity and the decrease of MDA content.Conclusion FTY720 exerts neuroprotective effects against the damage by cerebral ischemia/reperfusion probably through activating Nrf2/HO-1 signaling pathway.