Role of p53 in Neuroprotective Effects of Puerarin against MPP-induced Human Neuroblastoma SH-SY5Y Cell Death

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Role of p53 in Neuroprotective Effects of Puerarin against MPP-induced Human Neuroblastoma SH-SY5Y Cell Death

更新时间：2024-01-04

- Role of p53 in Neuroprotective Effects of Puerarin against MPP-induced Human Neuroblastoma SH-SY5Y Cell Death
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 20, Issue 4, Pages: 154-158(2014)
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- DOI：10.11653/syfj2014040154
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- Published：2014
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YAN Chun-lin, CHENG Yue-fa, WU Qing-wen, et al. Role of p53 in Neuroprotective Effects of Puerarin against MPP-induced Human Neuroblastoma SH-SY5Y Cell Death[J]. Chinese journal of experimental traditional medical formulae, 2014, 20(4): 154-158.
DOI：

YAN Chun-lin, CHENG Yue-fa, WU Qing-wen, et al. Role of p53 in Neuroprotective Effects of Puerarin against MPP-induced Human Neuroblastoma SH-SY5Y Cell Death[J]. Chinese journal of experimental traditional medical formulae, 2014, 20(4): 154-158. DOI： 10.11653/syfj2014040154.

摘要

目的：探讨1-甲基-4-苯基-吡啶离子（MPP+）诱导人神经母细胞瘤（SH-SY5Y）细胞凋亡过程中p53及其上下游相关基因的变化情况，并探讨葛根素干预的保护作用。方法：体外培养SH-SY5Y细胞株，待细胞密度达到70%融合后，用终浓度分别为50，100，150 μmol·L-1的葛根素预先作用3 h后加入1 mmol·L-1的MPP+继续培养，第48 h采用噻唑蓝（MTT）法测定细胞存活率，第24 h采用Annexin-V/PI流式细胞分析术检测细胞凋亡率，实时荧光定量PCR（RT-PCR）技术分析p53及其相关基因mRNA表达量变化。结果：MTT结果显示葛根素预处理的细胞比单纯MPP+处理的细胞存活率有显著性增加（P<0.05），说明葛根素对MPP+诱导的SH-SY5Y细胞具有保护作用且呈一定的剂量依赖性；Annexin-V/PI流式细胞分析术检测结果表明葛根素可以降低MPP+诱导的SH-SY5Y细胞的凋亡率；RT-PCR结果显示MPP+诱导可以增加SH-SY5Y细胞的p53 mRNA表达；葛根素预处理下调了p53及其相关基因p21，Bax，Caspase-3 的mRNA的表达，上调了p53负调节因子MDM2和凋亡抑制基因Bcl-2的mRNA的表达。结论：MPP+诱导可以导致SH-SY5Y细胞的p53 mRNA增加；葛根素可以通过调节p53及其相关基因的表达对MPP+诱导的SH-SY5Y细胞发挥保护作用。

Abstract

Objective: To investigate the expression changes of p53 mRNA and its related genes in 1-methyl-4-phenylpyridinium iodide(MPP+)-induced SH-SY5Y cells and observe the protective effects of puerarin against the cell death. Method: SH-SY5Y cells were incubated to 70% confluence

the medium containing 50

100

150 μmol·L-1 puerarin was added to SH-SY5Y cells

at 3 h after drug application 1 mmol·L-1 MPP+ was added to culture. The viability of MPP+-induced SH-SY5Y cells under different doses of puerarin was assayed by 3-(4

5-dimethylthiazol-2-yl)-2

5-diphenyltetrazolium bromide(MTT)at 48 h. The apoptosis rates were detected by flow cytometry with Annexin-V/PI fluorescence staining. The expression of p53 mRNA and its related genes was assayed by real-time quantitative reverse transcription-PCR(RT-PCR)at 24 h. Result: The puerarinpretreatment could apparently increase the viability and inhibit the early apoptosis of the MPP+-induced SH-SY5Y cells in the experiment(P<0.05). Annexin-V/PI fluorescence staining results showed that puerarin could reduce the apoptosis rate of MPP+-induced SH-SY5Y cells. Real-time PCR results showed that the MPP+ could increase the expression of p53 mRNA

but the expression of p53

p21

Bax

Caspase-3 mRNA was down-regulated in puerarin pretreatment groups compared with the MPP+ group

and the expression of MDM2 and Bcl-2 mRNA was up-regulated simultaneously. Conclusion: The expression of p53 mRNA in SH-SY5Y cells can increase by MPP+ and puerarin can regulate the p53 and its related genes and play a protective role against the death of MPP+-induced SH-SY5Y cells.

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Related Institution

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