丹参酮ⅡA抑制HT22海马神经元发生铁死亡的机制研究  

Research on the mechanism of tanshinone ⅡA inhibiting ferroptosis in HT22 hippocampus cells

作　　者：许璐 汤其强 Xu Lu;Tang Qiqiang(Dept of Neurology,The Affiliated Provincial Hospital of Anhui Medical University,Hefei230001)

机构地区：[1]安徽医科大学附属省立医院神经内科

出　　处：《安徽医科大学学报》2019年第6期833-839,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81573807)

摘　　要：目的探讨丹参酮ⅡA(TanⅡA)抑制小鼠海马细胞(HT22)发生铁死亡的作用机制。方法采用铁死亡激活剂Erastin处理HT22细胞建立铁死亡损伤模型,再用TanⅡA干预损伤的HT22细胞。以HT22细胞为研究对象,实验分4组:对照组、Erastin组、TanⅡA组,TanⅡA+锌原卟啉(ZnPP)组。采用CCK-8法检测细胞增殖;超氧化物阴离子荧光探针(DHE)、氟硼二吡咯(BODIPYTM)581/591C11检测细胞内活性氧(ROS)及脂质过氧化(LPO)变化;特异性Fe^2+荧光探针(FeRhoNoxTM-1)检测细胞内活性铁含量;免疫荧光检测细胞内铁蛋白表达;Western blot法检测各组细胞内血红素氧合酶1(HO-1)蛋白表达。结果0.5μmol/LErastin作用HT22细胞能明显抑制细胞活力(P<0.01),增加细胞内ROS、LPO及活性铁含量(P<0.001);与Erastin组比较,TanⅡA组细胞活力明显增加(P<0.001),细胞内ROS、LPO及活性铁含量明显减少。Westernblot结果显示TanⅡA能够促进细胞内HO-1蛋白的表达。结论TanⅡA对Erastin诱导细胞发生铁死亡具有保护作用,其机制可能是通过增加细胞内HO-1的表达,减少细胞内ROS、LPO、活性铁含量,从而对细胞发生铁死亡发挥保护作用。Objective To investigate the mechanism of tanshinone ⅡA(Tan ⅡA) inhibiting ferroptosis in HT22 cells. Methods HT22 cells were treated with Erastin to establish ferroptosis damage model,then Tan ⅡA was added to intervene in damage model.HT22 cells were divided into four groups: control,Erastin,Tan ⅡA and Tan ⅡA+ZnPP groups.Furthermore CCK-8 assay was performed to measure the cell survival rate;the content of intracellular reactive oxygen species(ROS),lipid peroxidation and ferrous iron were detected by Superoxide anion fluorescent probe(DHE),BODIPYTM 581/591 C11 and FeRhoNoxTM-1 probe respectively;and the expression of HO-1and ferritin were determined by Western blot and Immunofluorescence assay respectively. Results Compare with control group,treatment of HT22 cells with 0.5 μmol/L Erastin for 8 h resulted in significantly cell death( P <0.01),and the content of intracellular ROS,lipid peroxidation and ferrous iron increased sigificantly( P <0.001).Compared with the Erastin group,the survival rate increased significantly in Tan ⅡA group( P <0.001),meanwhile the content of intracellular ROS,lipid peroxidation and ferrous iron decreased significantly.Western blot showed that Tan ⅡA could promote the expression level of HO-1 protein in HT22 cells. Conclusion Tan ⅡA exert elicit protective effect on HT22 cells during ferroptosis damage induced by Erastin through increasing the expression of HO-1 and reducing the content of intracellular ROS,lipid peroxidation and ferrous iron.

关 键 词：丹参酮ⅡA Erastin HT22细胞 铁死亡 神经保护 HO-1 

分 类 号：R741.02[医药卫生—神经病学与精神病学]