铁对过表达α-突触核蛋白PC12细胞Rab35蛋白表达的影响  

EFFECT OF IRON ON THE PROTEIN EXPRESSION OF RAB35 IN PC12 CELLS WITH α-SYNUCLEIN OVEREXPRESSION

作　　者：李云 谢俊霞 宋宁 LI Yun;XIE Junxia;SONG Ning(Department of Physiology and Pathophysiology,School of Basic Medicine,Qingdao University Medical College,Qingdao 266071,China)

机构地区：[1]青岛大学医学部基础医学院生理学与病理生理学系,山东青岛266071

出　　处：《青岛大学学报（医学版）》2021年第2期202-205,共4页Journal of Qingdao University(Medical Sciences)

基　　金：国家自然科学基金面上项目(31871049)。

摘　　要：目的探究枸橼酸铁铵(FAC)对过表达α-突触核蛋白PC12细胞Rab35蛋白表达的影响。方法用多西环素(DOX)处理24 h诱导PC12细胞高表达α-突触核蛋白。应用100μmol/L和1 mmol/L FAC处理24 h后用免疫印迹法检测细胞内Rab35蛋白表达,探讨高铁对Rab35蛋白表达影响。进一步用FAC(100μmol/L)、铁死亡诱导剂erastin(10μmol/L)+FAC、铁死亡抑制剂ferrostatin-1(10μmol/L)+FAC处理细胞24 h,检测Rab35蛋白表达,探讨铁死亡水平对Rab35蛋白表达的影响。结果100μmol/L和1 mmol/L FAC处理过表达α-突触核蛋白PC12细胞不影响Rab35蛋白表达水平(F=1.601,q=1.726、2.466,P>0.05)。100μmol/L FAC处理组Rab35蛋白表达水平与对照组相比没有明显变化(F=13.35,q=0.442,P>0.05);FAC和erastin共处理组Rab35蛋白表达水平与对照组和FAC组相比均没有明显变化(q=3.415、2.973,P>0.05);但FAC和ferrostatin-1共处理组Rab35蛋白表达水平下调,与对照组和FAC组相比差异具有统计学意义(q=7.933、7.491,P<0.05)。结论铁对过表达α-突触核蛋白PC12细胞Rab35蛋白表达没有影响;铁死亡抑制剂在高铁情况下可降低Rab35蛋白表达,提示铁死亡可能参与了铁对Rab35蛋白的调控。Objective To investigate the effect of ferric ammonium citrate(FAC)on the protein expression of Rab35 in PC12 cells withα-synuclein overexpression.Methods PC12 cells were treated with doxycycline for 24 h to induce the high expression ofα-synuclein.After treatment with FAC at a dose of 100μmol/L or 1 mmol/L for 24 h,Western blotting was used to measure the protein expression of Rab35,and the effect of high iron level on the protein expression of Rab35 was analyzed.The cells were treated with FAC(100μmol/L),the ferroptosis inducer erastin(10μmol/L)+FAC,or the ferroptosis inhibitor ferrostatin-1(10μmol/L)+FAC for 24 h,and the protein expression of Rab35 was measured to investigate the effect of ferroptosis on the protein expression of Rab35.Results The treatment of PC12 cells withα-synuclein overexpression with 100μmol/L or 1 mmol/L FAC did not affect the protein expression level of Rab35(F=1.601;q=1.726,2.466;P>0.05).Compared with the control group,the 100μmol/L FAC treatment group had no significant change in the protein expression level of Rab35(F=13.35,q=0.442,P>0.05);compared with the control group and the FAC group,the FAC+erastin treatment group had no significant change in the protein expression level of Rab35(q=3.415,2.973;P>0.05);compared with the control group and the FAC group,the FAC+ferrostatin-1 treatment group had a significant reduction in the protein expression level of Rab35(q=7.933,7.491;P<0.05).Conclusion Iron has no effect on the protein expression of Rab35 in PC12 cells withα-synuclein overexpression,and with the presence of high iron level,ferroptosis inhibitor can reduce the protein expression of Rab35,suggesting that ferroptosis might be involved in the regulation of Rab35 protein by iron.

关 键 词：铁 PC12细胞 铁死亡 rab GTP结合蛋白质类 

分 类 号：R338.2[医药卫生—人体生理学]