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

发布时间：2018-12-25 09:31

【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(P0.05),丁苯酞组和URMC-099组细胞存活率高于MPP+组(P0.05);MPP+组细胞凋亡率高于对照组(P0.05),丁苯酞组和URMC-099组细胞凋亡率低于MPP+组(P0.05);与对照组相比,MPP+组p-MLK3、p-JNK蛋白表达量增加(P0.05),p-ERK1/2蛋白表达量降低(P0.05);与MPP+组相比,丁苯酞组和URMC-099组p-MLK3、p-JNK蛋白表达量降低(P0.05),p-ERK1/2蛋白表达量升高(P0.05)。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。
[Abstract]:Objective to investigate the effect of butylphthalide on the mixed lineage kinase 3 (MLK3) pathway induced by 1-methyl-4-phenyl-pyridine ion (MPP) in SH-SY5Y cells and its mechanism of cell proliferation and apoptosis. Methods SH-SY5Y cells in logarithmic growth phase were divided into control group, MPP group, butyrophthalide group and URMC-099 group. The control group was cultured for 24 h, MPP group was cultured with 1 mmol/L MPP for 24 h, butylphthalide group was pretreated with 10 渭 mol/L butylphthalide for 3 h. URMC-099 group was pretreated with 200nmol/L MLK3 pathway specific inhibitor URMC-099 for 3 h and MPP for 24 h. Cell morphology was observed by inverted phase contrast microscope, cell activity was detected by thiazolyl blue colorimetry, apoptosis rate was detected by Annexin-V/PI double staining flow cytometry, and MLK3 phosphorylated protein (p-MLK3) was detected by, Western blotting by Hoechst33342 fluorescence staining. Expression of c-Jun amino-terminal kinase phosphorylation protein (p-JNK) and extracellular regulated protein kinase phosphorylation protein (p-ERK1/2). Results the cell survival rate of MPP group was lower than that of control group (P0.05). The cell survival rate of butyphthalide group and URMC-099 group was higher than that of MPP group (P0.05). The apoptosis rate of MPP group was higher than that of control group (P0.05), and that of butyphthalide group and URMC-099 group was lower than that of MPP group (P0.05). Compared with the control group, the expression of p-MLK3PJNK protein in MPP group was increased (P0.05), and the expression of p-ERK1/2 protein was decreased (P0.05). Compared with MPP group, butyphthalide group and URMC-099 group decreased the expression of p-MLK _ 3, p-JNK protein (P0.05), and increased the expression of p-ERK1/2 protein (P0.05). Conclusion Butylphthalide can reduce the apoptosis and promote the proliferation of SH-SY5Y cells induced by MPP. The mechanism may be that butyphthalide can regulate the expression of p-JNKP p-ERK 1 / 2 protein downstream by inhibiting the MLK3 pathway.
【作者单位】：华北理工大学护理与康复学院;华北理工大学冀唐学院;
【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)
【分类号】：R742.5

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