白藜芦醇通过激活p53/SLC7A11/GPX4信号通路抑制铁死亡减少脑缺血再灌注损伤

Resveratrol Pathway Activates p53/SLC7A11/GPX4 Signaling Pathway to Inhibit Ferroptosis and Improve Cerebral Ischemia-reperfusion Injury

目的探讨白藜芦醇对脑缺血再灌注后铁死亡的抑制作用及其机制。方法250~280 g的SPF级SD雄性大鼠63只,随机分为假手术组、脑缺血再灌注组(MCAO/R)组、白藜芦醇组。利用线栓法建立大鼠疾病模型。给药组剂量为30 mg/kg。给药28 d后应用Zea-longa评分评估大鼠神经功能。给药24 h取材大鼠脑组织,通过称重法评估脑水肿情况,应用免疫荧光染色检测谷胱甘肽过氧化酶4(glutathione peroxidase 4,GPX4),长链脂酰辅酶A合成酶4(long chain lipoyl coenzyme A synthetase 4,ACSL4)。试剂盒检测各组脑组织中Fe^(2+)、谷胱甘肽过氧化物酶(GSH)、超氧化物歧化酶(superoxide dismutase,SOD)和丙二醛(malonaldehyde,MDA)水平。利用蛋白免疫印记(Western Blot)对不同组别的大鼠脑组织中p53、SLC7A11以及GPX4蛋白表达进行检测。结果与损伤组相比,白藜芦醇治疗组Zea-longa评分和脑组织水含量、Fe^(2+)、MDA明显降低,GSH、SOD表达明显增加;免疫荧光染色结果显示,GPX4表达明显增强、ACSL4表达降低。WB通路蛋白结果显示,白藜芦醇治疗组P53表达降低,SLC7A11表达升高。结论白藜芦醇通过调控p53/SLC7A11/GPX4信号通路抑制MCAO/R大鼠铁死亡,进而促进缺血再灌注大鼠神经功能恢复。

Objective To investigate the inhibitory effect of resveratrol on iron death after cerebral ischemia reperfusion and its mechanism.Methods 63 male sprague-dawley rats,weighing 250~280 g,were randomly divided into sham-operated group,MCAO/R group and resveratrol group.The rat disease model was established by using thread-plug method.The dosage of the treated group was 30mg/kg.Zea-longa score was used to evaluate the neurological function of rats 28 days after administration.The brain tissues of rats were collected at 24 h after administration.The brain edema was evaluated by weighing method.The levels of Glutathione peroxidase 4(GPX4)and long chain lipoyl coenzyme A synthetase 4(ACSL4)were detected by immunofluorescence staining.The levels of Fe^(2+),glutathione peroxidase(GSH),superoxide dismutase(SOD)and malonaldehyde(MDA)in the brain issues of each group were detected by the kit.The protein expressions of p53,SLC7A11 and GPX4 in different groups of rat brain were detected by Western Blot.Results Compared with the injured group,the Zea-longa score,the water content,Fe^(2+)and MDA in the brain,and the expression of GSH and SOD in the resveratrol-treated group were significantly decreased,and the results of immunofluorescence staining showed that the content of GSH and SOD in the brain were significantly increased in the resveratrol-treated group,the expression of GPX4 was increased and the expression of ACSL4 was decreased.The results of WB pathway showed that the expression of P53 was decreased and the expression of SLC7A11 was increased in resveratrol treated group.Conclusion Resveratrol inhibits iron death in MCAO/R rats by regulating the p53/SLC7A11/GPX4 signaling pathway,thereby promoting the recovery of neural function after ischemia/reperfusion.