Telmisartan attenuated LPS-induced neuroinflammation in human IMR-32 neuronal cell line via SARM in AT1R independent mechanism
Telmisartan attenuated LPS-induced neuroinflammation in human IMR-32 neuronal cell line via SARM in AT1R independent mechanism
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DOI：
10.1016/j.lfs.2015.03.005
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发表时间：
2015-06-01
期刊：
影响因子：
6.1
通讯作者：
Ramanathan, Muthiah
中科院分区：
文献类型：
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作者：
Saravanan, Prathab Balaji;Shanmuganathan, Muthusamy V.;Ramanathan, Muthiah
目的：探讨替米沙坦（TS）对LPS中毒神经细胞的保护作用，并探讨其可能的神经保护机制。主要方法：设计TLR4和AT1R特异性引物，用rtPCR方法确认受体在IMR-32和Neuro2A细胞系中的表达。MTT法检测TS的保护作用。通过Western blotting检测TLR4特异性接头蛋白SARM和MyD88的表达和激活，磷酸化NF κ B、PPAR γ、MAPK p38、c-JNK、ERR的表达和激活，阐明TS的作用机制。选择性PPAR γ拮抗剂GW9662被用来证实PPAR γ激活与TLR4介导的NF κ B炎症机制之间的联系。ELISA法检测促炎细胞因子TNF α、IL1 β、IL-6及抗炎细胞因子110的释放。主要发现：IMR-32细胞表达TLR4受体，Neuro2A细胞同时表达AT1R和TLR4受体。TS对LPS中毒的两种细胞系均有显著保护作用。TS通过激活PPAR γ和SARM蛋白显著抑制TLR4介导的炎症反应，选择性PPAR γ拮抗剂GW9662显著逆转这一作用，证实PPAR γ激活与TLR4通过SARM介导的炎症反应存在联系。意义：LPS中毒的人神经元IMR-32细胞是研究TLR4受体表达引起的炎症介导的神经退行性变的良好体外模型。我们的研究强烈建议，PPAR γ激活TS的多效性作用是在LPS诱导的TLR4介导的炎症中通过SARM接头蛋白在IMR-32细胞系中起保护作用的原因。(C) 2015 Elsevier Inc .版权所有
Aim: The aim of this study was to find the protective role of Telmisartan (TS) in LPS intoxicated neuronal cells and elucidate the possible neuroprotective mechanism of action.Main methods: TLR4 and AT1R specific primers were designed and used in rtPCR to confirm the receptor expression in IMR-32 and Neuro2A cell lines. The protective effect of TS was assayed by MTT assay. The mechanism of action of TS was elucidated by assessing the expression and activation of TLR4 specific adaptor proteins SARM and MyD88, phosphorylated NF kappa B, PPAR gamma, MAPK p38, c-JNK, ERR by Western blotting. Selective PPAR gamma antagonist GW9662 was used to confirm the link between PPAR gamma activation and TLR4 mediated NF kappa B inflammatory mechanisms. The pro-inflammatory cytokines TNF alpha, IL1 beta, and IL-6 and anti-inflammatory cytokine 110 release were measured by ELISA.Key findings: IMR-32 cells expressed TLR4 receptor and Neuro2A cells expressed both AT1R and TLR4 receptors. TS significantly protected both the cell lines from LPS intoxication. TS significantly suppressed the TLR4 mediated inflammatory response by PPAR gamma and SARM protein activation and the effect was reversed significantly by selective PPAR gamma antagonist GW9662, confirming the existence of link between PPAR gamma activation and TLR4 mediated inflammation via SARM.Significance: LPS intoxicated human neuronal IMR-32 cells can be a good in vitro model to study inflammatory mediated neurodegeneration due to TLR4 receptor expression. Our study strongly recommends that the PPAR gamma activating pleiotropic effect of TS is responsible for the protective effect in LPS induced TLR4 mediated inflammation via SARM adaptor protein in the IMR-32 cell line. (C) 2015 Elsevier Inc All rights reserved.