丁苯酞通过混合谱系激酶3信号通路对1-甲基-4-苯基-吡啶离子诱导的SH-SY5Y细胞增殖和凋亡的影响  

Effects of DL-3-n-Butylphthalide on Proliferation and Apoptosis of 1-Methyl-4-Phenylpyridinium-induced SH-SY5Y Cells via Mixed Lineage Kinase 3 Signaling Pathway

作　　者：郭子梦 吴庆文 陈秀秀 关亚丽 李鹏飞[2] 王妍[2] 程月发[2] GUO Zi-meng;WU Qing-wen;CHEN Xiu-xiu;GUAN Ya-li;LI Peng-fei;WANG Yan;CHENG Yue-fa(College of Nursing and Rehabilitation, North China University of Science and Technology, Tangshan, Hebei 063210, China;Jitang College of North China University of Science and Technology, Tangshan, Hebei 063210, China)

机构地区：[1]华北理工大学护理与康复学院,河北唐山市063210 [2]华北理工大学冀唐学院,河北唐山市063210

出　　处：《中国康复理论与实践》2017年第11期1284-1289,共6页Chinese Journal of Rehabilitation Theory and Practice

基　　金：华北理工大学研究生创新项目(No.2017S37);华北理工大学博士科研启动基金项目(No.35647699);河北省高等学校科学研究青年基金项目(No.QN201722)

摘　　要：目的探讨丁苯酞对1-甲基-4-苯基-吡啶离子(MPP+)诱导的SH-SY5Y细胞混合谱系激酶3(MLK3)通路的影响,及其对细胞增殖与凋亡的作用机制。方法对数生长期SH-SY5Y细胞分为对照组、MPP+组、丁苯酞组和URMC-099组,对照组正常培养,MPP+组加1 mmol/L MPP+培养24 h,丁苯酞组予10μmol/L丁苯酞预处理3 h后,加入MPP+培养24 h,URMC-099组予200nmol/L MLK3通路特异性抑制剂URMC-099预处理3 h后,加入MPP+培养24 h。倒置相差显微镜观察细胞形态,噻唑蓝比色法检测细胞活性,Annexin-V/PI双染流式细胞术检测细胞凋亡率,Hoechst33342荧光染色法观察凋亡细胞,Western blotting检测MLK3磷酸化蛋白(p-MLK3)、c-Jun氨基末端激酶磷酸化蛋白(p-JNK)和细胞外调节蛋白激酶磷酸化蛋白(p-ERK1/2)的表达。结果 MPP+组细胞存活率低于对照组(P<0.05),丁苯酞组和URMC-099组细胞存活率高于MPP+组(P<0.05);MPP+组细胞凋亡率高于对照组(P<0.05),丁苯酞组和URMC-099组细胞凋亡率低于MPP+组(P<0.05);与对照组相比,MPP+组p-MLK3、p-JNK蛋白表达量增加(P<0.05),p-ERK1/2蛋白表达量降低(P<0.05);与MPP+组相比,丁苯酞组和URMC-099组p-MLK3、p-JNK蛋白表达量降低(P<0.05),p-ERK1/2蛋白表达量升高(P<0.05)。结论丁苯酞可减少MPP+诱导的SH-SY5Y细胞凋亡,促进细胞增殖,其机制可能是通过抑制MLK3通路,调节下游p-JNK、p-ERK1/2蛋白表达。Objective To investigate the effects of DL-3-n-Butylphthalide(NBP)on proliferation and apoptosis of1-methyl-4-phenylpyridinium(MPP+)-induced SH-SY5Y cells,and mechanisms via mixed lineage kinase3(MLK3)signaling pathway.Methods The SH-SY5Y cells were divided into control group,MPP+group,NBP group and URMC-099group,that cultured normally,with1mmol/L MPP+for24hours,with10μmol/L NBP for3hours and then with MPP+for24hours,and with200nmol/L MLK3inhibitor URMC-099for3hours and then with MPP+for24hours,respectively.The morphology of SH-SY5Y cells was observed under inverted phase contrast microscope and the survival rate was measured with3-(4,5-Cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays.The apoptosis was quantified under flow cytometry with Annexin V/PI fluorescence staining,and the nuclear morphology was observed with Hoechst33342staining.The expression of phosphorylated protein of MLK3(p-MLK3),c-Jun N-terminal kinase(p-JNK),extra cellular regulated protein kinases(p-ERK1/2)were detected with Western blotting.Results Compared with the control group,the survival rate reduced and apoptosis increased in MPP+group(P<0.05),with the increase of p-MLK3and p-JNK and decrease of p-ERK1/2d(P<0.05).Compared with MPP+group,the survival rate increased and apoptosis reduced in both NBP and URMC-099groups(P<0.05),with the decrease of p-MLK3and p-JNK and increase of p-ERK1/2(P<0.05).Conclusion NBP can decrease the apoptosis and promote the proliferation of SH-SY5Y cells induced by MPP+,which may be associated with inhibiting MLK3signaling pathway,and regulating the downstream p-JNK and p-ERK1/2.

关 键 词：帕金森病 丁苯酞 混合谱系激酶3 凋亡 SH-SY5Y细胞 

分 类 号：R742.5[医药卫生—神经病学与精神病学]