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 (12): 2147-2155.doi: 10.4103/1673-5374.241470

• 原著:退行性病与再生 • 上一篇    下一篇

下调信号转导和转录激活因子3表达可加剧NLRP3炎性小体介导神经元的氧化应激损伤

  

  • 收稿日期:2018-08-03 出版日期:2018-12-15 发布日期:2018-12-15
  • 基金资助:

    贵州省科技厅项目（[2016]1131），贵州省卫计委项目（2015-326），中国国家自然科学基金项目不发达地区专项基金（81560482），贵州省创新团队研究基金（KY[2016]033）

Downregulation of signal transduction and STAT3 expression exacerbates oxidative stress mediated by NLRP3 inflammasome

Hua Bai1, 2, Qi-Fang Zhang3, Juan-Juan Duan3, De-Jun Yu1, Li-Jie Liu2   

  1. 1 Medical Laboratory Center, Third Affiliated Hospital, Guizhou Medical University, Duyun, Guizhou Province, China
    2 Department of Neurology, Third Affiliated Hospital, Guizhou Medical University, Duyun, Guizhou Province, China
    3 Key Laboratory of Endemic and Ethnic Diseases of Ministry of Education, and Key Laboratory of Medical Molecular Biology, Guizhou Medical University, Guiyang, Guizhou Province, China
  • Received:2018-08-03 Online:2018-12-15 Published:2018-12-15
  • Contact: Hua Bai, PhD, baihcd@126.com; Qi-Fang Zhang PhD, abcde9981@21cn.com.
  • Supported by:

    This study was supported by Department of Science and Technology in Guizhou Province of China, No. Basic [2016]1131 (to Qian-Ke-He; to HB); Department of Health and Family Planning Commission in Guizhou Province of China, No. 2015-326 (to HB); Less Developed Regions of the National Natural Science Foundation of China, No. 81560482; the Research Foundation for Creative Research Groups of Education Bureau of Guizhou Province of China, No. KY[2016]033 (to QFZ).

PDF

220

摘要：

活化NLRP3炎症小体可能通过氧化应激和神经源性炎症参与阿尔茨海默病的发病，信号转导和转录激活因子3基因低表达在一定程度上可促进神经退行性疾病的发生。为验证实验设计程序：(1)SHSY5Y细胞加入1 mM H2O2进行氧化应激损伤，检验人细胞特异性信号转导和转录激活因子3-shRNA沉默信号转导和转录激活因子3的表达，同时以Ca2+螯合剂BAPATA-AM(0.1mM)预处理30min的细胞作为对照；(2)以Western blot分析caspase-1、NLRP3和信号转导和转录激活因子3的表达，以ELISA分析培养基中白细胞介素1β水平，以流式细胞仪计算凋亡细胞数量；(3)发现过氧化氢诱导可促进NLRP3炎性小体活化，降低信号转导和转录激活因子3丝氨酸727的磷酸化；(4)BAPATA-AM预处理可消除过氧化氢诱导的NLRP3炎症小体活化、caspase-1表达、白细胞介素1β表达和细胞凋亡，而抑制信号转导和转录激活因子3基因表达BAPATA-AM的作用失效；(5)上述结果数据表明，信号转导和转录激活因子3表达下调可能会增强NLRP3介导的氧化应激作用，这可能不依赖于Ca2+信号通路。

orcid:0000-0002-5617-0215(Hua Bai)
        0000-0002-6708-9439(Qi-Fang Zhang)

关键词: 信号转导和转录激活因子3, 钙离子, caspase-1, NLRP3, 炎性小体, 过氧化氢, 阿尔茨海默病, shRNA, SHSY5Y细胞, 神经再生

Abstract:

Activated nucleotide binding to the oligonucleotide receptor protein 3 (NLRP3) inflammasome is possibly involved in the pathogenesis of Alzheimer’s disease through oxidative stress and neurogenic inflammation. Low expression of the signal transducer and activator of transcription 3 (STAT3) gene may promote the occurrence of neurodegenerative diseases to some extent. To clarify the roles of the NLRP3 inflammasome and STAT3 expression in oxidative stress, (1) SHSY5Y cells were incubated with 1 mM H2O2 to induce oxidative stress injury, and the expression of human-cell-specific signal transduction, STAT3-shRNA silencing signal transduction and STAT3 were detected. Cells were pretreated with Ca2+ chelator BAPATA-AM (0.1 mM) for 30 minutes as a control. (2) Western blot assay was used to analyze the expression of caspase-1, NLRP3, signal transduction and STAT3. Enzyme-linked immunosorbent assay was used to analyze interleukin-1β levels. Flow cytometry was carried out to calculate the number of apoptotic cells. We found that H2O2 treatment activated NLRP3 inflammasomes and decreased phosphorylation of signal transduction and STAT3 serine 727. BAPTA-AM pretreatment abolished the H2O2-induced activation of NLRP3 inflammasomes, caspase-1 expression, interleukin-1β expression and apoptosis in SHSY5Y cells, and had no effect in cells with downregulated STAT3 expression by RNAi. The findings suggest that downregulation of signal transduction and STAT3 expression may enhance the oxidative stress mediated by NLRP3, which may not depend on the Ca2+ signaling pathway.

Key words: nerve regeneration, signal transducer and activator of transcription 3, calcium, caspase-1, nucleotide binding to the oligonucleotide receptor protein 3, inflammasome, hydrogen peroxide, Alzheimer’s disease, shRNA, SHSY5Y cells, neural regeneration