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
计量
- 文章访问数: 489
- HTML全文浏览量: 319
- PDF下载量: 7
- 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

下载: 全尺寸图片幻灯片

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

下载: 全尺寸图片幻灯片

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

下载: 全尺寸图片幻灯片

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

下载: 全尺寸图片幻灯片

参考文献(20)

[1]	Jankovic J. Parkinson's disease: clinical features and diagnosis[J]. J Neurol Neurosurg Psychiatry,2008,79(4):368-376. doi: 10.1136/jnnp.2007.131045
[2]	Prajjwal P,Flores Sanga H S,Acharya K,et al. Parkinson's disease updates: Addressing the pathophysiology,risk factors,genetics,diagnosis,along with the medical and surgical treatment[J]. Ann Med Surg (Lond),2023,85(10):4887-4902. doi: 10.1097/MS9.0000000000001142
[3]	Medeiros M S,Schumacher-Schuh A,Cardoso A M,et al. Iron and oxidative stress in parkinson's disease: An observational study of injury biomarkers[J]. PLoS One,2016,11(1):e0146129. doi: 10.1371/journal.pone.0146129
[4]	Romuk E,Szczurek W,Nowak P,et al. Effects of propofol on oxidative stress parameters in selected parts of the brain in a rat model of parkinson disease[J]. Postepy Hig Med Dosw (Online),2016,70(0):1441-1450.
[5]	Kalia L V,Lang A E. Parkinson's disease[J]. Lancet,2015,386(9996):896-912. doi: 10.1016/S0140-6736(14)61393-3
[6]	Alarcon-Gil J,Sierra-Magro A,Morales-Garcia J A,et al. Neuroprotective and anti-inflammatory effects of linoleic acid in models of parkinson's disease: The implication of lipid droplets and lipophagy[J]. Cells,2022,11(15):2297. doi: 10.3390/cells11152297
[7]	Yao Y,Liao C,Qiu H,et al. Effect of eleutheroside e on an MPTP-induced parkinson's disease cell model and its mechanism[J]. Molecules,2023,28(9):3820. doi: 10.3390/molecules28093820
[8]	Ju D T,Sivalingam K,Kuo W W,et al. Effect of vasicinone against paraquat-induced MAPK/p53-mediated apoptosis via the IGF-1R/PI3K/AKT pathway in a parkinson's disease-associated SH-SY5Y cell model[J]. Nutrients,2019,11(7):1655. doi: 10.3390/nu11071655
[9]	Han X,Han B,Zhao Y,et al. Rosmarinic acid attenuates rotenone-induced neurotoxicity in sh-sy5y parkinson's disease cell model through abl inhibition[J]. Nutrients,2022,14(17):3508. doi: 10.3390/nu14173508
[10]	Cai L J,Tu L,Li T,et al. Up-regulation of microRNA-375 ameliorates the damage of dopaminergic neurons,reduces oxidative stress and inflammation in Parkinson's disease by inhibiting SP1[J]. Aging (Albany NY),2020,12(1):672-689. doi: 10.18632/aging.102649
[11]	Motawi T K,Al-Kady R H,Abdelraouf S M,et al. Empagliflozin alleviates endoplasmic reticulum stress and augments autophagy in rotenone-induced Parkinson's disease in rats: Targeting the GRP78/PERK/eIF2α/CHOP pathway and miR-211-5p[J]. Chem Biol Interact,2022,362:110002. doi: 10.1016/j.cbi.2022.110002
[12]	Ayaz M,Anwar F,Saleem U,et al. Parkinsonism attenuation by antihistamines via downregulating the oxidative stress,histamine,and inflammation[J]. ACS Omega,2022,7(17):14772-14783. doi: 10.1021/acsomega.2c00145
[13]	Salaramoli S,Amiri H,Joshaghani H R,et al. Bio-synthesized selenium nanoparticles ameliorate Brain oxidative stress in Parkinson disease rat models[J]. Metab Brain Dis,2023,38(6):2055-2064. doi: 10.1007/s11011-023-01222-6
[14]	Zamanian M Y,Parra R M R,Soltani A,et al. Targeting Nrf2 signaling pathway and oxidative stress by resveratrol for Parkinson's disease: An overview and update on new developments[J]. Mol Biol Rep,2023,50(6):5455-5464. doi: 10.1007/s11033-023-08409-1
[15]	Yu W,Gao D,Jin W,et al. Propofol prevents oxidative stress by decreasing the ischemic accumulation of succinate in focal cerebral ischemia-reperfusion injury[J]. Neurochem Res,2018,43(2):420-429. doi: 10.1007/s11064-017-2437-z
[16]	Yang Y,Xia Z,Xu C,et al. Ciprofol attenuates the isoproterenol-induced oxidative damage,inflammatory response and cardiomyocyte apoptosis[J]. Front Pharmacol,2022,13:1037151. doi: 10.3389/fphar.2022.1037151
[17]	Sun L,Dou X,Yang W. Propofol protects rats against intra-cerebroventricular streptozotocin-induced cognitive dysfunction and neuronal damage[J]. Folia Morphol (Warsz),2023,82(2):248-255. doi: 10.5603/FM.a2022.0027
[18]	Gonullu E,Dagistan G,Erkin Y,et al. Demonstration of the protective effect of propofol in rat model of cisplatin-induced neuropathy[J]. Bratisl Lek Listy,2023,124(1):64-69.
[19]	Wang S,Song T,Leng C,et al. Propofol protects against the neurotoxicity of 1-methyl-4-phenylpyridinium[J]. Mol Med Rep,2016,13(1):309-314. doi: 10.3892/mmr.2015.4570
[20]	高青,秦燕,任晓亮,等. 基于Nrf2/RAGE/NF-κB信号通路探究丙泊酚对帕金森病大鼠认知障碍的影响[J]. 西部医学,2023,35(12):1745-1750. doi: 10.3969/j.issn.1672-3511.2023.12.006

相关文章(20)

[1]	张宁丽, 周臣, 思永玉, 欧阳杰, 林岚, 陈莹, 牛钦, 李鸿. 艾司氯胺酮复合丙泊酚在无痛人流术中的应用, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230514
[2]	董丽, 孙士波, 孙曙光. 灯盏花乙素抗氧化应激机制在防治心脑血管疾病中的研究进展, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20220423
[3]	张丹妮, 段霞光, 郝春光, 支小军, 朱海峰, 黄再青. 氢吗啡酮复合丙泊酚对大鼠结直肠扩张模型的麻醉效果及机制, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210705
[4]	张宁平, 张琼, 张宁丽, 思永玉, 屈启才. 不同剂量丙泊酚减少老年患者胃肠镜检查术中依托咪酯所致肌阵挛的临床研究, 昆明医科大学学报.
[5]	曾庆菊, 李慧, 王晋文. 褪黑素对糖尿病大鼠氧化应激及足细胞凋亡的影响及作用机制, 昆明医科大学学报.
[6]	魏星. 个体化靶控输注丙泊酚在重度烧伤患者麻醉中的临床应用, 昆明医科大学学报.
[7]	曾柏瑞. 甲基苯丙胺与HIV-Tat蛋白协同改变大鼠血脑屏障通透性的氧化应激作用机制, 昆明医科大学学报.
[8]	屈启才. 舒芬太尼复合靶控输注丙泊酚静脉诱导对高血压患者血流动力学的影响, 昆明医科大学学报.
[9]	张美琼. 氯胺酮和瑞芬太尼分别伍用丙泊酚用于无痛肠镜的临床比较, 昆明医科大学学报.
[10]	方育. 丙泊酚靶控输注麻醉诱导时复合芬太尼对患者效应室丙泊酚浓度和血流动力学的影响, 昆明医科大学学报.
[11]	丁艳杰. 类风湿合并颈动脉硬化患者血清氧化应激状态与MMPs相关性分析, 昆明医科大学学报.
[12]	刘松. 1 800 MHz电磁波对大鼠心肌氧化应激的影响, 昆明医科大学学报.
[13]	范丹. 七氟烷、丙泊酚麻醉对心脏瓣膜置换手术患者心肌损伤标志物影响的研究, 昆明医科大学学报.
[14]	张媛. 1 800 MHz电磁辐射对大鼠皮肤组织氧化应激的影响, 昆明医科大学学报.
[15]	李建钢. 依托咪酯与丙泊酚全麻诱导对老年患者心血管稳定性的比较研究, 昆明医科大学学报.
[16]	闫庆峰. 木犀草素对冷保存大鼠心脏心功能及氧化应激反应的影响, 昆明医科大学学报.
[17]	陈学强. 靶控输注丙泊酚对高原高血压病人全麻诱导时血压变异性的影响, 昆明医科大学学报.
[18]	. 丙泊酚复合小剂量咪达唑仑、芬太尼、氯胺酮在绝育术中的临床应用, 昆明医科大学学报.
[19]	. 丙泊酚复合全麻加气管表面麻醉用于声带手术的临床观察, 昆明医科大学学报.
[20]	桂莉. 2型糖尿病大鼠骨骼肌氧化应激与胰岛素抵抗的关系, 昆明医科大学学报.