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 / 中国康复理论与实践

ABSTRACT

Objective To investigate the effects of DL-3-n-Butylphthalide(NBP)on proliferation and apoptosis of 1-methyl-4-phenyl-pyridinium (MPP +)-induced SH-SY5Y cells, and mechanisms via mixed lineage kinase 3 (MLK3) signaling pathway. Methods The SH-SY5Y cells were divided into control group,MPP+group,NBP group and URMC-099 group,that cultured normally,with 1 mmol/L MPP+for 24 hours,with 10μmol/L NBP for 3 hours and then with MPP+for 24 hours,and with 200 nmol/L MLK3 inhibitor URMC-099 for 3 hours and then with MPP+for 24 hours,respectively.The morphology of SH-SY5Y cells was observed under inverted phase contrast mi-croscope and the survival rate was measured with 3-(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 Hoechst 33342 staining.The expression of phosphorylated protein of MLK3(p-MLK3),c-Jun N-terminal kinase(p-JNK),extra cellular regulated protein ki-nases(p-ERK1/2)were detected with Western blotting.Results Compared with the control group,the survival rate reduced and apoptosis in-creased in MPP+group(P<0.05),with the increase of p-MLK3 and p-JNK and decrease of p-ERK1/2 d(P<0.05).Compared with MPP+group,the survival rate increased and apoptosis reduced in both NBP and URMC-099 groups(P<0.05),with the decrease of p-MLK3 and 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 in-duced by MPP+,which may be associated with inhibiting MLK3 signaling pathway,and regulating the downstream p-JNK and p-ERK1/2.