[1]甘敏,周巧雅,洪美静,等.肝X受体激动剂通过激活SIRT1信号级联改善Aβ诱导的视网膜色素上皮细胞炎症和凋亡反应[J].第三军医大学学报,2020,42(14):1398-1406.
 GAN Min,ZHOU Qiaoya,HONG Meijing,et al.Liver X receptor agonist ameliorates amyloid β-induced inflammation and apoptosis in retinal pigment epithelial cells by activating SIRT1 signaling cascade[J].J Third Mil Med Univ,2020,42(14):1398-1406.
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肝X受体激动剂通过激活SIRT1信号级联改善Aβ诱导的视网膜色素上皮细胞炎症和凋亡反应(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第14期
页码:
1398-1406
栏目:
神经科学
出版日期:
2020-07-30

文章信息/Info

Title:
Liver X receptor agonist ameliorates amyloid β-induced inflammation and apoptosis in retinal pigment epithelial cells by activating SIRT1 signaling cascade
作者:
甘敏周巧雅洪美静李雪彭惠
重庆医科大学附属第一医院眼科,眼科学重庆市市级重点实验室,重庆市眼科研究所
Author(s):
GAN Min ZHOU Qiaoya HONG Meijing LI Xue PENG Hui

Department of Ophthalmology,  Chongqing Eye Institute, Chongqing Key Laboratory of Ophthalmology, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
视网膜色素上皮肝X受体沉默信息调节因子1淀粉样&beta年龄相关性黄斑变性
Keywords:
 pigment epithelium liver X receptor silencing information regulator 1 &beta-amyloid age-related macular degeneration
分类号:
R329.25; R774.5; R977.6
文献标志码:
A
摘要:

目的探讨LXR-SIRT1调控轴在淀粉样β(amyloidβ, Aβ)诱导的视网膜上皮(retinal pigment epithelial, RPE)细胞炎症和凋亡中的作用。方法体外培养ARPE-19细胞系,收集湿性年龄相关性黄斑变性(age-related macular degeneration,AMD)患者和正常受试者的人外周血单个核细胞(human peripheral blood mononuclear cell, PBMC)并检测其中沉默信息调节因子1(silencing information regulator 1, SIRT1)的mRNA表达。用1.0、5.0 μmol/L的Aβ1-40和5.0 μmol/L Aβ40-1处理ARPE-19细胞24 h,Real-time PCR检测SIRT1、IL-1β、p21在ARPE-19细胞中的表达,探讨Aβ对ARPE-19细胞炎症、凋亡的影响。用5.0 μmol/L Aβ1-40和5.0 μmol/L TO90处理ARPE-19细胞,Western blot检测SIRT1、LXRα、ace-NF-κB、NF-κB、ace-p53、p53、p21的表达,研究Aβ1-40刺激及TO90预处理对SIRT1信号级联的影响。在敲低SIRT1后,Western blot检测NF-κB和p53信号通路的表达,进一步验证TO90对SIRT1信号级联的调控作用。结果SIRT1在湿性AMD患者的PBMC中明显下调(P<0.01)。Aβ1-40刺激ARPE-19细胞后,SIRT1在mRNA及蛋白水平明显下调(P<0.05),p21在mRNA及蛋白水平明显上调(P<0.05),IL-1β在基因水平明显上调(P<0.05),ace-NF-κB、NF-κB、ace-p53、p53在蛋白水平显著上调(P<0.05),而TO90预处理则可逆转Aβ1-40对这些因子的调节(P<0.05)。SIRT1敲低后,TO90对ace-NF-κB、NF-κB、ace-p53、p53上调的逆转作用显著下降(P<0.05)。结论LXR激动剂通过激活SIRT1信号级联改善Aβ诱导的ARPE-19细胞炎症及凋亡反应。
 

Abstract:

ObjectiveTo investigate the role of liver X receptor (LXR)-SIRT1 regulatory axis in inflammation and apoptosis of retinal pigment epithelial (RPE) cells induced by β-amyloid (Aβ). MethodsHuman peripheral blood mononuclear cells (PBMCs) were collected from patients with wet age-related macular degeneration (AMD) and normal subjects for detection of SIRT1 mRNA expression. And then, human RPE cell line ARPE-19 stimulated with 1.0, 5.0 μmol/L Aβ1-40 or 5.0 μmol/L Aβ40-1 was examined for expression of SIRT1, IL-1β and p21 using real-time PCR to investigate the effects of Aβ on cell inflammation and apoptosis; Subsequently, the expression levels of SIRT1, LXRα, ace-NF-κB, NF-κB, ace-p53, p53, and p21 proteins in ARPE-19 cells treated with 5.0 μmol/L Aβ1-40, 5.0 μmol/L TO90, or both were detected using Western blotting to analyze the effect of Aβ1-40 stimulation and TO90 pretreatment on SIRT1 signaling cascade. Finally, after knocking down SIRT1, the expression of NF-κB and p53 signaling pathways was detected by Western blot analysis to further verify the regulatory effect of TO90 on the SIRT1 signaling cascade. ResultsSIRT1 was significantly down-regulated in the PBMCs of patients with wet AMD (P<0.05). Aβ stimulation of ARPE-19 cells caused obvious downregulation of SIRT1 (P<0.05) and upregulation of p21 at both the mRNA and protein levels (P<0.05), upregulation of IL-1β at the mRNA level (P<0.05), and upregulation of ace-NF-κB, NF-κB, ace-p53, and p53 at the protein level (P<0.05); TO90 pretreatment effectively abolished the effects of Aβ on these factors (P<0.05). After SIRT1 knockdown, the reverse effect of TO90 on the upregulation of ace-NF-κB, NF-κB, ace-p53, and p53 was significantly reduced (P<0.05). ConclusionLXR agonists improve Aβ-induced inflammation and apoptosis in ARPE-19 cells by activating the SIRT1 signaling cascade.
 

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更新日期/Last Update: 2020-07-23