溶血磷脂酸联合6-羟基多巴胺对SH-SY5Y细胞凋亡的诱导作用及其机制

doi:10.13481/j.1671-587X.20260116

Abstract

Abstract:

Objective To discuss the inductive effect of lysophosphatidic acid （LPA） combined with 6-hydroxydopamine （6-OHDA） on the apoptosis of SH-SY5Y cells， and to clarify its related mechanism. Methods The Parkinson’s disease （PD） cell model was established by using 100 μmol·L^-1 6-OHDA； the SH-SY5Y cells， PC12 cells and N2a cells were selected and divided into control group （incomplete medium only）， 4 μmol·L^-1 LPA group， 100 μmol·L^-1 6-OHDA group， 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group， 10 μmol·L^-1 LPA group， and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group， and treated for 24 h. Methylthiazolydiphenyl-tetrazolium bromide（MTT） method was used to detect the activities of the SH-SY5Y cells in various groups； annexin Ⅴ fluorescein isothiocyanate（Annexin Ⅴ-FITC）/propidium iodide（PI） staining and flow cytometry were used to detect the apoptotic rates of the SH-SY5Y cells in various groups； Western blotting method was used to detect the expression levels of Caspase-3 related proteins in three kinds of cells and the expression levels of B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， p38 mitogen-activated protein kinase （p38 MAPK）， phosphorylated p38 MAPK （p-p38 MAPK） proteins and LPA1 receptor protein in the SH-SY5Y cells in various groups. Results The MTT assay results showed that compared with 100 μmol·L?1 6-OHDA group， the numbers and activities of the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group were significantly decreased （P<0.05 or P<0.01）. The flow cytometry results showed that compared with 100 μmol·L^-1 6-OHDA group， the apoptotic rates of the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group were increased （P<0.05 or P<0.01）. The Western blotting results showed that compared with 100 μmol·L^-1 6-OHDA group， the expression levels of Caspase-3 protein in three kinds of cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group had no significant changes， and the differences were not statistically significant （P>0.05）； the expression levels of Cleaved Caspase-3 protein in the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group were significantly increased （P<0.01）； the expression levels of Cleaved Caspase-3 protein in the PC12 cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group were significantly increased （P<0.05 or P<0.01）； the expression level of Cleaved Caspase-3 protein in the N2a cells in 4 μmol·L?1 LPA+100 μmol·L?1 6-OHDA group was increased （P<0.05）. The Western blotting results showed that compared with 100 μmol·L^-1 6-OHDA group， the expression levels of Bax and Bcl-2 proteins in the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L?1 6-OHDA group and 10 μmol·L?1 LPA+100 μmol·L?1 6-OHDA group had no significant changes， and the differences were not statistically significant （P>0.05）， but the Bax/Bcl-2 ratio was increased （P<0.05）； the expression level of p38 MAPK protein in the SH-SY5Y cells in 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group was increased （P<0.05）； the expression level of p-p38 MAPK protein in the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group was significantly increased （P<0.01）， and the p-p38 MAPK/p38 MAPK ratio was increased （P<0.05）； the expression level of LPA1 receptor protein in the SH-SY5Y cells in 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group was decreased （P<0.05）. Conclusion The combined application of LPA and 6-OHDA can significantly induce the apoptosis of SH-SY5Y cells， and its mechanism may be related to the up-regulated expression levels of p38 MAPK and p-p38 MAPK proteins in the cells.

Key words: Lysophosphatidic acid, 6-hydroxydopamine, SH-SY5Y cell, Apoptosis, p38 mitogen-activated protein kinase

CLC Number:

Q545

Yaping LI,Chunyan TAN,Lujie ZHAO,Jiayi ZHAO,Ting LI,Xiao YANG,Xiaoyun YANG. Inductive effect of lysophosphatidic acid combined with 6-hydroxydopamine on apoptosis of SH-SY5Y cells and its mechanism[J].Journal of Jilin University(Medicine Edition), 2026, 52(1): 152-161.

Figures/Tables 7

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References 34

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Group	Cell activity
Control	100.00±1.35
4 μmol·L^-1 LPA	84.98±2.86
100 μmol·L^-1 6-OHDA	83.89±3.20
4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA	69.19±3.40^**
10 μmol·L^-1 LPA	74.65±3.51
10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA	71.14±3.50^*

Inductive effect of lysophosphatidic acid combined with 6-hydroxydopamine on apoptosis of SH-SY5Y cells and its mechanism

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