中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (3): 440-448.doi: 10.4103/1673-5374.228726

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

替勃龙预处理对调节臭氧暴露大鼠海马Tau磷酸化的激酶和磷酸酶的影响

  

  • 收稿日期:2018-01-13 出版日期:2018-03-15 发布日期:2018-03-15

Effect of tibolone pretreatment on kinases and phosphatases that regulate the expression and phosphorylation of Tau in the hippocampus of rats exposed to ozone

Rodolfo Pinto-Almazán1, 2, 3, Julia J. Segura-Uribe4, 5, Marvin A. Soriano-Ursúa5, Eunice D. Farfán-García5, Juan M. Gallardo6, Christian Guerra-Araiza7   

  1. 1 Unidad de Investigación Hospital Regional de Alta Especialidad Ixtapaluca, Carretera Federal México-Puebla km 34.5, C.P. 56530. Ixtapaluca,State of Mexico, Mexico
    2 Institute for the Developing Mind, Children’s Hospital Los Angeles, Los Angeles, CA, USA
    3 Department of Pediatrics, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA
    4 Unidad de Investigación Médica en Enfermedades Neurológicas, Hospital de Especialidades, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Av. Cuauhtémoc 330 Col. Doctores. C. P. 06720. Mexico City, Mexico
    5 Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón, Col. Casco de Santo Tomás. C. P. 11340. Mexico City, Mexico
    6 Unidad de Investigación Médica en Enfermedades Nefrológicas, Hospital de Especialidades, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Av. Cuauhtémoc 330 Col. Doctores. C. P. 06720. Mexico City, Mexico
    7 Unidad de Investigación Médica en Farmacología, Hospital de Especialidades, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Av. Cuauhtémoc 330 Col. Doctores. C. P. 06720. Mexico City, Mexico
  • Received:2018-01-13 Online:2018-03-15 Published:2018-03-15
  • Contact: Christian Guerra-Araiza, Ph.D.,christianguerra2001@gmail.com.
  • Supported by:

    This study was supported by FIS/IMSS project No. FIS/IMSS/PROT/G09/751 and FIS/IMSS/PROT/G11-2/1013.

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摘要：

氧化应激参与神经退行性疾病的发病。替勃龙是一种用于治疗绝经期症状的合成激素，可降低脂质过氧化水平，预防臭氧（O3）所致学习和记忆功能障碍。然而，目前尚不清楚替勃龙是否能阻止氧化应激诱导的微管相关蛋白Tau磷酸化增加。为观察替勃龙不同时间给药对雄性大鼠臭氧暴露模型Tau磷酸化、糖原合成酶激酶-3β活化及Akt和蛋白磷酸酶2A和第10号染色体上磷酸酶及张力蛋白同源缺失的基因蛋白（PTEN）磷酸酶失活的影响。实验将大鼠随机分为10组：对照组（未暴露于臭氧中），对照+替勃龙组（未暴露于臭氧+ 灌胃替勃龙1 mg/kg）; 暴露于臭氧7，15，30和60d的臭氧暴露组和暴露于臭氧前给予7，15，30和60d的替勃龙1 mg/kg灌胃干预组。采用蛋白质印迹法检测海马Tau，糖原合成酶激酶3β，Akt，蛋白磷酸酶2A和PTEN总蛋白和磷酸化蛋白，采用分光光度法检测海马氧化应激标记物超氧化物歧化酶活性和硝基酪氨酸、丙二醛水平。实验观察到臭氧暴露增加了大鼠海马Tau磷酸化蛋白水平，这与蛋白磷酸酶2A、PTEN、Akt、糖原合成酶激酶3β蛋白水平降低有关。臭氧暴露15和60d，海马丙二醛和硝基酪氨酸水平增加，并且超氧化物歧化酶活性降低。替勃龙长期给药（15d以上）可阻止以上臭氧暴露的影响。这些结果表明，用替勃龙有益于预防臭氧暴露引起的神经变性。

orcid:0000-0002-7164-4116(Christian Guerra-Araiza)

 

关键词: 氧化应激, 大气暴露室, Tau, GSK3, 海马, 神经保护剂, 神经再生

Abstract:

Oxidative stress (OS) is a key process in the development of many neurodegenerative diseases, memory disorders, and other pathological processes related to aging. Tibolone (TIB), a synthetic hormone used as a treatment for menopausal ymptoms, decreases lipoperoxidation levels, prevents memory impairment and learning disability caused by ozone (O3) exposure. However, it is not clear if TIB could prevent the increase in phosphorylation induced by oxidative stress of the microtubule-associated protein Tau. In this study, the effects of TIB at different times of administration on the phosphorylation of Tau, the activation of glycogen synthase kinase-3β (GSK3β), and the inactivation of Akt and phosphatases PP2A and PTEN induced by O3 exposure were assessed in adult male Wistar rats. Rats were divided into 10 groups: control group (ozone-free air plus vehicle [C]), control + TIB group (ozone-free air plus TIB 1 mg/kg [C + TIB]); 7,15, 30, and 60 days of ozone exposure groups [O3] and 7, 15, 30, and 60 days of TIB 1 mg/kg before ozone exposure groups [O3 + TIB]. The effects of O3 exposure and TIB administration were assessed by western blot analysis of total and phosphorylated Tau, GSK3β, Akt, PP2A, and PTEN proteins and oxidative stress marker nitrotyrosine, and superoxide dismutase activity and lipid peroxidation of malondialdehyde by two different spectrophotometric methods (Marklund and TBARS, respectively). We observed that O3 exposure increases Tau phosphorylation, which is correlated with decreased PP2A and PTEN protein levels, diminished Akt protein levels, and increased GSK3β protein levels in the hippocampus of adult male rats. The effects of O3 exposure were prevented by the long-term treatment (over 15 days) with TIB. Malondialdehyde and nitrotyrosine levels increased from 15 to 60 days of exposure to O3 in comparison to C group, and superoxide dismutase activity decreased. Furthermore, TIB administration limited the changes induced by O3 exposure. Our results suggest a beneficial use of hormone replacement therapy with TIB to prevent neurodegeneration caused by O3 exposure in rats.

Key words: tibolone, oxidative stress, ozone exposure, Tau, GSK3, hippocampus, neuroprotection