lncRNA-BC200调控Aβ25-35诱导的神经细胞PC12炎症因子表达和细胞凋亡
作者:
作者单位:

(1.桂林医学院附属医院神经内科,广西桂林市 541001;2.桂林医学院,广西桂林市 541000)

作者简介:

梁静,硕士,副主任医师,研究方向为神经免疫及变性疾病,E-mail为ljliang1w@163.com。

基金项目:

广西自然科学基金项目(2015GXNSFBA139135)


LncRNA-BC200 regulating the expression of inflammatory factor and apoptosis of neuronal cell PC12 induced by Aβ25-35
Author:
Affiliation:

1.Neurology Department, Affiliated Hospital of Guilin Medical University, Guilin, Guangxi 541001, China;2.Guilin Medical University, Guilin, Guangxi 541000, China)

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    摘要:

    目的 探讨lncRNA-BC200对Aβ25-35诱导的神经细胞炎症和细胞凋亡的影响及可能机制。方法 将神经细胞PC12分为正常对照组(细胞常规培养)和10、20、40 μmol/L Aβ25-35组(分别用10、20、40 μmol/L Aβ25-35干预细胞24 h),流式细胞术检测细胞凋亡,qRT-PCR法检测细胞中lncRNA-BC200表达。将PC12细胞分为正常对照组、Aβ25-35组(用20 μmol/L Aβ25-35干预PC12细胞24 h)、si-NC+Aβ25-35组(用20 μmol/L Aβ25-35干预转染si-NC的PC12细胞24 h)、si-lncRNA-BC200+Aβ25-35组(用20 μmol/L Aβ25-35干预转染si-lncRNA-BC200的PC12细胞24 h)和TNF-α+si-lncRNA-BC200+Aβ25-35组[用20 μmol/L Aβ25-35和20 μg/L肿瘤坏死因子α(TNF-α)共同干预转染si-lncRNA-BC200的PC12细胞24 h],流式细胞术检测细胞凋亡,酶联免疫吸附法检测细胞培养上清液中TNF-α、白细胞介素6(IL-6)和γ干扰素(IFN-γ)表达,Western blot检测细胞中cleaved-Caspase-3、p-p65和p-IκBα蛋白表达。结果 10、20、40 μmol/L Aβ25-35组PC12细胞凋亡率和lncRNA-BC200表达均高于正常对照组。Aβ25-35组细胞中cleaved-Caspase-3、p-p65和p-IκBα蛋白表达,以及TNF-α、IL-6和IFN-γ水平均高于正常对照组。si-lncRNA-BC200+Aβ25-35组PC12细胞凋亡率和细胞中cleaved-Caspase-3、p-p65和p-IκBα蛋白表达及TNF-α、IL-6和IFN-γ水平均低于Aβ25-35组。TNF-α+si-lncRNA-BC200+Aβ25-35组PC12细胞凋亡率和细胞中cleaved-Caspase-3、p-p65和p-IκBα蛋白表达及TNF-α、IL-6和IFN-γ水平均高于si-lncRNA-BC200+Aβ25-35组。结论 敲减lncRNA-BC200可能通过抑制NF-κB信号通路的激活减少Aβ25-35诱导的神经细胞PC12分泌炎症因子及细胞凋亡。

    Abstract:

    Aim To investigate the effect of lncRNA-BC200 on the inflammation and apoptosis of nerve cells induced by Aβ25-35 and its possible mechanism. Methods The nerve cells PC12 were divided into NC group (conventional cell culture) and 0,0, 40 μmol/L Aβ25-35 groups (cells were treated with 0,0, 40 μmol/L Aβ25-35 for 24 h), and then cell apoptosis was detected by flow cytometry. qRT-PCR method was used to detect the expression of lncRNA-BC200 in cells. The PC12 cells were divided into NC group (cells were routinely cultured), Aβ25-35 group (PC12 cells were intervened with 20 μmol/L Aβ25-35 for 24 h), si-NC+Aβ25-35 group (PC12 cells were transfected with si-NC and then intervened with 20 μmol/L Aβ25-35 for 24 h), si-lncRNA-BC200+Aβ25-35 group (PC12 cells were transfected with si-lncRNA-BC200 and then intervened with 20 μmol/L Aβ25-35 for 24 h) and TNF-α+si-lncRNA-BC200+Aβ25-35 group (PC12 cells were transfected with si-lncRNA-BC200 and then co-intervened with 20 μmol/L Aβ25-35 and 20 μg/L tumor necrosis factor (TNF-α) for 24 h). Flow cytometry was used to detect cell proliferation activity and apoptosis, ELISA was used to detect the expression of TNF-α, interleukin-6 (IL-6) and interferon-γ (IFN-γ) in the cell culture supernatant, and Western blot was used to detect the protein expression of cleaved-Caspase-3, p-p65 and p-IκBα in the cells. Results The apoptosis rate of PC12 cells and the expression of lncRNA-BC200 were higher in 0,0, 40 μmol/L Aβ25-35 groups than NC group. The protein expression of cleaved-Caspase-3, p-p65 and p-IκBα and the levels of TNF-α, IL-6 and IFN-γ in Aβ25-35 group cells were all higher than NC group. The apoptotic rate of PC12 cells, the protein expression of cleaved-Caspase-3, p-p65 and p-IκBα, and the levels of TNF-α, IL-6 and IFN-γ were all lower in the si-lncRNA-BC200+Aβ25-35 group than Aβ25-35 group. The apoptotic rate of PC12 cells, the protein expression of cleaved-Caspase-3, p-p65 and p-IκBα, and the levels of TNF-α, IL-6 and IFN-γ in the TNF-α+si-lncRNA-BC200+Aβ25-35 group were all higher than the si-lncRNA-BC200+Aβ25-35 group. Conclusion Knockdown of lncRNA-BC200 may inhibit Aβ25-35-induced neuronal cell PC12 secretion of inflammatory factors and apoptosis by inhibiting the activation of NF-κB signaling pathway.

    参考文献
    [1] KE S, YANG Z, YANG F, et al.Long noncoding RNA NEAT1 aggravates Aβ-induced neuronal damage by targeting miR-107 in alzheimer's disease.Yonsei Med J, 9,0(7):640-650.
    [2] WANG Q, GE X, ZHANG J, et al.Effect of lncRNA WT1-AS regulating WT1 on oxidative stress injury and apoptosis of neurons in Alzheimer's disease via inhibition of the miR-375/SIX4 axis.Aging (Albany NY), 0,2(23):23974-23995.
    [3] LEE S, YOUN K, JUN M.Major compounds of red ginseng oil attenuate Aβ(25-35)-induced neuronal apoptosis and inflammation by modulating MAPK/NF-κB pathway.Food Funct, 8,9(8):4122-4134.
    [4] 李晓峰, 张文瑛, 梁铁生.老年痴呆症患者外周血中LncRNA-BC200和LncRNA-17 A水平及应用价值.中风与神经疾病杂志, 9,6(9):790-793.
    [5] LIU J, ZUO X, HAN J, et al.MiR-9-5p inhibits mitochondrial damage and oxidative stress in AD cell models by targeting GSK-3β.Biosci Biotechnol Biochem, 0,4(11):2273-2280.
    [6] QU J, XIONG X, HUJIE G, et al.MicroRNA-132-3p alleviates neuron apoptosis and impairments of learning and memory abilities in Alzheimer's disease by downregulation of HNRNPU stabilized BACE1.Cell Cycle, 1,0(21):2309-2320.
    [7] HABAIKE A, YAKUFU M, CONG Y, et al.Neuroprotective effects of Fomes officinalis Ames polysaccharides on Aβ(25-35)-induced cytotoxicity in PC12 cells through suppression of mitochondria-mediated apoptotic pathway.Cytotechnology, 0,2(4):539-549.
    [8] 雷敏, 吴丽荣, 刘英.氯沙坦对大鼠脑缺血再灌注损伤的保护作用及分子机制.中国动脉硬化杂志, 0,8(3):213-218.
    [9] LEI S, WU S, WANG G, et al.Pinoresinol diglucoside attenuates neuroinflammation, apoptosis and oxidative stress in a mice model with Alzheimer's disease.Neuroreport, 1,2(3):259-267.
    [10] 梁春荣, 刘雨辉, 王叶冉, 等.阿尔茨海默病患者外周血炎症因子水平与认知功能的相关性研究.解放军医学杂志, 4,9(2):133-137.
    [11] DONG L X, ZHANG Y Y, BAO H L, et al.LncRNA NEAT1 promotes Alzheimer's disease by down regulating micro-27a-3p.Am J Transl Res, 1,3(8):8885-8896.
    [12] ZHANG Y Y, BAO H L, DONG L X, et al.Silenced lncRNA H19 and up-regulated microRNA-129 accelerates viability and restrains apoptosis of PC12 cells induced by Aβ(25-35) in a cellular model of Alzheimer's disease.Cell Cycle, 1,0(1):112-125.
    [13] WANG Q B, GE X M, ZHANG J, et al.Effect of lncRNA WT1-AS regulating WT1 on oxidative stress injury and apoptosis of neurons in Alzheimer's disease via inhibition of the miR-375/SIX4 axis.Aging(Albany NY), 0,2(23):23974-23995
    [14] YI J P, CHEN B, YAO X X, et al.Upregulation of the lncRNA MEG3 improves cognitive impairment, alleviates neuronal damage, and inhibits activation of astrocytes in hippocampus tissues in Alzheimer's disease through inactivating the PI3K/Akt signaling pathway.J Cell Biochem, 9,0(8):18053-18065.
    [15] MA P, LI Y, ZHANG W, et al.Long non-coding RNA MALAT1 inhibits neuron apoptosis and neuroinflammation while stimulates neurite outgrowth and its correlation with miR-125b mediates PTGS2, CDK5 and FOXQ1 in Alzheimer's disease.Curr Alzheimer Res, 9,6(7):596-612.
    [16] TAN N, ZHU B, SHU H, et al.Effect of lncRNABC200 on proliferation and migration of liver cancer cells in vitro and in vivo.Oncol Rep, 0,3(2):461-470.
    [17] GAO B B, WANG S X.LncRNA BC200 regulates the cell proliferation and cisplatin resistance in non-small cell lung cancer via PI3K/AKT pathway.Eur Rev Med Pharmacol Sci, 9,3(3):1093-1101.
    [18] ZHAO R, CAO X, JIN S, et al.LncRNA BC200 promotes esophageal squamous cell cancer migration and invasion and can regulate ATF4 expression.Front Oncol, 0,0:1392.
    [19] 王瑶, 杨超, 邓克廷, 等.血清长链非编码RNA BC200水平在阿尔茨海默病诊疗中的应用价值.华中科技大学学报(医学版), 8,7(5):589-593.
    [20] KONG F, SUN Y, SONG W, et al.miR-216a alleviates LPS-induced acute lung injury via regulating JAK2/STAT3 and NF-κB signaling.Hum Cell, 0,3(1):67-78.
    [21] 张政军, 郝钰, 万婷婷.沙格列汀对ox-LDL诱导的血管内皮细胞损伤及miR-590/TLR4/NF-κB表达的影响.中国动脉硬化杂志, 9,7(11):938-943.
    [22] JIN X, LIU M Y, ZHANG D F, et al.Baicalin mitigates cognitive impairment and protects neurons from microglia-mediated neuroinflammation via suppressing NLRP3 inflammasomes and TLR4/NF-κB signaling pathway.CNS Neurosci Ther, 9,5(5):575-590.
    [23] HU W, WEN L, CAO F, et al.Down-regulation of miR-107 worsen spatial memory by suppressing SYK expression and inactivating NF-κB signaling pathway.Curr Alzheimer Res, 9,6(2):135-145.
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梁静,陈汉仁,蒋静子,毛敏芸,彭天婵. lncRNA-BC200调控Aβ25-35诱导的神经细胞PC12炎症因子表达和细胞凋亡[J].中国动脉硬化杂志,2022,30(5):403~409.

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  • 收稿日期:2021-09-16
  • 最后修改日期:2021-11-30
  • 在线发布日期: 2022-05-10