丙泊酚调节MPP+诱导的SH-SY5Y细胞线粒体氧化应激和凋亡

谭莹; 秦海燕; 孙翔; 苏彦伊; 王英宝

doi:10.12259/j.issn.2095-610X.S20240305

丙泊酚调节MPP+诱导的SH-SY5Y细胞线粒体氧化应激和凋亡

doi: 10.12259/j.issn.2095-610X.S20240305

1.
昆明医科大学第一附属医院麻醉科，云南昆明　650032
2.
昆明市延安医院泌尿外科，云南昆明　650051

基金项目: 昆明市卫生科技人才培养基金资助项目[2023-sw（后备）-18]

详细信息

作者简介:
谭莹（1986～），女，云南瑞丽人，医学硕士，主治医师，主要从事麻醉与心脑保护研究工作

通讯作者:
王英宝，E-mail：944450200@qq.com

中图分类号: R742.5
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- 被引次数: 0
出版历程
- 收稿日期: 2023-12-25
- 网络出版日期: 2024-03-11
- 刊出日期: 2024-03-30

Propofol Regulates MPP+-induced Mitochondrial Oxidative Stress and Apoptosis in SH-SY5Y Cells

1.
Dept. of Anesthesiology，The 1st Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650032
2.
Dept. of Urology，Yan’an Hospital of Kunming City，Kunming Yunnan 650051，China

摘要

摘要: 目的探讨不同浓度的丙泊酚（propofol，PPF）对1-甲基-4-苯基吡啶离子（1-Methyl-4-phenylpyridinium，MPP+）诱导的帕金森（Parkinson's disease, PD）模型细胞氧化应激和凋亡的作用。方法利用1 mM MPP+诱导人神经母细胞瘤细胞SH-SY5Y构建PD细胞模型。PPF处理组分别用10、20、40、80 μM PPF在MPP+诱导前刺激SH-SY5Y细胞4 h。细胞计数法（cell counting kit-8，CCK-8）评估细胞增殖活力。2′，7′-二氯二氢荧光素二乙酸酯（H2DCF-DA）荧光探针用于检测细胞中活性氧（reactive oxygen species，ROS）。试剂盒分别检测丙二醛（malondialdehyde，MDA）和还原型烟酰胺腺嘌呤二核苷酸磷酸（reduced nicotinamide adenine dinucleotide phosphate oxidase，NADPH）氧化酶的水平。Western blot用于检测线粒体和细胞质中细胞色素c的蛋白表达以及细胞中Bcl-2、Bax和Cleaved caspase-3蛋白表达。流式细胞术分析细胞凋亡率。结果 MPP+诱导明显抑制SH-SY5Y细胞的增殖（P < 0.001），促进细胞中ROS（P < 0.001）、MDA（P < 0.001）、NADPH氧化酶（P < 0.01）、细胞质中细胞色素c的表达（P < 0.01）并诱导细胞凋亡（P < 0.001）以及促凋亡蛋白Bax和Cleaved caspase-3的表达（P < 0.01），减少线粒体中细胞色素c的蛋白表达（P < 0.01）和抗凋亡蛋白Bcl-2的表达（P < 0.01）。PPF预处理可缓解MPP+诱导引起的SH-SY5Y细胞增殖抑制以及氧化应激和凋亡的促进作用（P < 0.001），且40 μM和80 μM对细胞的作用更为显著。结论 PPF预处理可缓解MMP+诱导的SH-SY5Y细胞氧化应激，减少细胞凋亡。
- 帕金森 /
- 氧化应激 /
- 1-甲基-4-苯基吡啶离子 /
- 丙泊酚
Abstract: Objective To investigate the effects of different concentrations of PPF on oxidative stress and apoptosis of PD model cells induced by MPP+. Methods The human neuroblastoma cell SH-SY5Y was induced by 1 mM MPP+ to establish PD cell model. In PPF treatment group, SH-SY5Y cells were stimulated with 10, 20, 40 and 80 μM PPF for 4 h before MPP+ induction. Cell counting kit-8（CCK-8） was performed to evaluate cell proliferation activity. H2DCF-DA fluorescent probe was used to detect ROS in cells. The levels of MDA and NADPH oxidase were analyzed by the kit. Western blot examined the protein expression of cytochrome c in mitochondria and cytoplasm, as well as the relative expression of Bcl-2, Bax and cleaved caspase-3 in SH-SY5Y cells. Apoptosis rate was analyzed by flow cytometry. Results MPP+ significantly inhibited the proliferation of SH-SY5Y cells（P < 0.001）, promoted the level of ROS（P < 0.001）, MDA（P < 0.001）, NADPH oxidase（P < 0.01）, cytochrome c in cytoplasm（P < 0.01） and induced apoptosis（P < 0.001） and the relative expression of pro-apoptosis protein Bax and cleaved caspase-3（P < 0.01）, reduced cytochrome c protein in mitochondria（P < 0.01） and the relative expression of anti-apoptosis protein（P < 0.01）. PPF pretreatment alleviated the proliferation inhibition, oxidative stress and apoptosis promotion of SH-SY5Y cells induced by MPP+（P < 0.001）, and the effects of 40 μM and 80 μM on cells were more significant. Conclusion PPF pretreatment can alleviate the oxidative stress of SH-SY5Y cells induced by MMP+ and reduce apoptosis rate.
- Parkinson's disease /
- Oxidative stress /
- MPP+ /
- Propofol

HTML全文

图 1 MPP+诱导对SH-SY5Y细胞线粒体氧化应激的作用

A：MPP+诱导对SH-SY5Y细胞增殖活力的作用；B：荧光探针检测ROS水平；C：ROS水平统计；D：试剂盒检测MDA水平；E：试剂盒检测NADPH氧化酶水平；与NC组比较，^**P < 0.01，^***P < 0.001。

Figure 1. Effect of MPP+ induction on mitochondrial oxidative stress in SH-SY5Y cells

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图 2 MPP+诱导对SH-SY5Y细胞中细胞色素C和细胞凋亡的影响

A：Western blot用于检测细胞色素c的蛋白表达；B：细胞色素c蛋白表达统计；C：细胞凋亡检测；D：细胞凋亡率统计；E：Western blot检测凋亡相关蛋白表达；F：凋亡相关蛋白表达统计；与NC组相比，^**P < 0.01，^***P < 0.001。

Figure 2. Effect of MPP+ induction on cytochrome c and cell apoptosis in SH-SY5Y cells

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图 3 PPF调控MPP+诱导的SH-SY5Y细胞线粒体氧化应激

A：不同浓度PPF对SH-SY5Y细胞增殖活力的作用；B：ROS荧光强度统计；C：荧光探针检测不同浓度PPF对ROS水平的影响；D：不同浓度PPF对细胞中MDA浓度的影响；E：不同浓度PPF对细胞中NADPH氧化酶浓度的影响；与MPP+组比较，^*P < 0.05，^**P < 0.01，^***P < 0.001。

Figure 3. PPF regulated MPP+-induced mitochondrial oxidative stress in SH-SY5Y cells

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图 4 PPF调控MPP+诱导的SH-SY5Y细胞中细胞色素c和细胞凋亡

A：Western blot检测线粒体及细胞质中细胞色素c的蛋白表达；B：细胞色素c蛋白表达统计；C：细胞凋亡率检测；D：细胞凋亡率统计；E：Western blot检测凋亡相关蛋白表达；F：凋亡相关蛋白表达统计；与MPP+组比较，^**P < 0.01，^***P < 0.001。

Figure 4. PPF regulated MPP+-induced cytochrome c and cell apoptosis in SH-SY5Y cells

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