白杨素通过抑制铁死亡减轻大鼠脑缺血再灌注损伤

Chrysin alleviates cerebral ischemia-reperfusion injury by inhibiting ferroptosis in rats

该研究旨在探讨白杨素是否通过抑制铁死亡,减轻大鼠脑缺血再灌注损伤。雄性SD大鼠随机分为假手术组、模型组、白杨素高剂量组(200 mg·kg^(-1))、白杨素中剂量组(100 mg·kg^(-1))、白杨素低剂量组(50 mg·kg^(-1))、阳性药组(金纳多,21.6 mg·kg^(-1))共6组。通过短暂性大脑中动脉闭塞(transient middle cerebral artery occlusion,tMCAO)制备脑缺血再灌注大鼠模型,术后24 h进行指标评定及取材。神经功能评分检测神经功能;2,3,5-三苯基氯化四氮唑(2,3,5-triphenyl tetrazolium chloride,TTC)法测定脑梗死面积;苏木素-伊红(hematoxylin-eosin,HE)染色和尼氏染色观察脑组织形态结构;普鲁士蓝染色观察脑组织铁聚集水平;生化试剂检测血清和脑组织总铁、脂质过氧化物、丙二醛含量;实时荧光定量聚合酶链式反应(real-time quantitative polymerase chain reaction,RT-qPCR)、免疫组化和Western blot检测脑组织溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)、转铁蛋白受体(transferrin receptor,TFR1)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long chain family member 4,ACSL4)、前列腺素内过氧化物合成酶2(prostaglandin-endoperoxide synthase 2,PTGS2)的mRNA和蛋白表达。与模型组相比,各给药组大鼠神经功能恢复,脑梗死率降低,脑组织病理学改变减轻。筛选得到白杨素低剂量组(50 mg·kg^(-1))为最佳给药剂量组。与模型组相比,白杨素组大鼠脑组织和血清的总铁、脂质过氧化物、丙二醛含量显著降低,SLC7A11、GPX4的mRNA表达量和蛋白表达量显著增加,TFR1、PTGS2、ACSL4的mRNA表达量和蛋白表达量显著减少。白杨素可能通过调节铁死亡相关靶点,进而调控铁代谢,抑制脑缺血再灌注导致的神经元铁死亡。

The purpose of this study is to investigate whether chrysin reduces cerebral ischemia-reperfusion injury(CIRI)by inhi-biting ferroptosis in rats.Male SD rats were randomly divided into a sham group,a model group,high-,medium-,and low-dose chrysin groups(200,100,and 50 mg·kg^(-1)),and a positive drug group(Ginaton,21.6 mg·kg^(-1)).The CIRI model was induced in rats by transient middle cerebral artery occlusion(tMCAO).The indexes were evaluated and the samples were taken 24 h after the operation.The neurological deficit score was used to detect neurological function.The 2,3,5-triphenyl tetrazolium chloride(TTC)staining was used to detect the cerebral infarction area.Hematoxylin-eosin(HE)staining and Nissl staining were used to observe the morphological structure of brain tissues.Prussian blue staining was used to observe the iron accumulation in the brain.Total iron,lipid pero-xide,and malondialdehyde in serum and brain tissues were detected by biochemical reagents.Real-time quantitative polymerase chain reaction(RT-qPCR),immunohistochemistry,and Western blot were used to detect mRNA and protein expression of solute carrier fa-mily 7 member 11(SLC7A11),transferrin receptor 1(TFR1),glutathione peroxidase 4(GPX4),acyl-CoA synthetase long chain family member 4(ACSL4),and prostaglandin-endoperoxide synthase 2(PTGS2)in brain tissues.Compared with the model group,the groups with drug intervention showed restored neurological function,decreased cerebral infarction rate,and alleviated pathological changes.The low-dose chrysin group was selected as the optimal dosing group.Compared with the model group,the chrysin groups showed reduced content of total iron,lipid peroxide,and malondialdehyde in brain tissues and serum,increased mRNA and protein expression levels of SLC7A11 and GPX4,and decreased mRNA and protein expression levels of TFR1,PTGS2,and ACSL4.Chrysin may regulate iron metabolism via regulating the related targets of ferroptosis and inhibit neuronal ferroptosis induced by CIRI.