绿原酸通过调控 MIR497HG/miR-29b-3p/SIRT1 轴预防低氧缺血环境中微胶质细胞诱导的神经元凋亡和氧化应激。
Chlorogenic Acid Prevents Microglia-Induced Neuronal Apoptosis and Oxidative Stress under Hypoxia-Ischemia Environment by Regulating the MIR497HG/miR-29b-3p/SIRT1 Axis.
机构信息
Central Laboratory, Affiliated Fuzhou First Hospital of Fujian Medical University, No. 190 Dadao Road, Fuzhou 350009, China.
Department of Neurology, Fujian Provincial Hospital, Shengli Clinical Medical College of Fujian Medical University, No.134 East Street, Fuzhou 350001, China.
出版信息
Dis Markers. 2022 May 25;2022:1194742. doi: 10.1155/2022/1194742. eCollection 2022.
BACKGROUND
Chlorogenic acid (CGA) is a polyphenolic compound with antioxidant and anti-inflammatory properties. CGA has been shown to improve neuroinflammation. This study is aimed at elucidating the exact mechanism by which CGA reduces neuroinflammation.
METHODS
Oxygen and glucose deprivation (OGD) was utilized to treat BV2 microglia and HT-22 hippocampal neurons to engineer an model of hypoxic ischemia reperfusion. The levels of inflammatory factors (IL-1, IL-6, TNF-, IL-4, and IL-10) and oxidative stress factors (MDA, SOD, and GSH-PX) in microglia were determined by ELISA kits. The neuron proliferation was assessed by CCK-8 assay, and LDH kit was used to determine LDH release in neurons. The fluorescent dye DCF-DA was employed to measure ROS levels in neurons. Correlation of MIR497HG, miR-29b-3p, and SIRT1/NF-B in neurons and microglia was determined by qRT-PCR. Expressions of inflammatory proteins (COX2, iNOS), oxidative stress pathways (Nrf2, HO-1), and apoptosis-related proteins (Bcl-2, Bax, caspase3, caspase8, and caspase9) in microglia or neurons were determined by western blot. The interactions between MIR497HG and miR-29b-3p, as well as between miR-29b-3p and SIRT1, were determined by dual luciferase assay and RIP assay.
RESULTS
CGA attenuated OGD-mediated inflammation and oxidative stress in microglia and inhibited microglia-mediated neuronal apoptosis. CGA increased the levels of MIR497HG and SIRT1 and suppressed the levels of miR-29b-3p in BV2 and HT-22 cells. MIR497HG knockdown, miR-29b-3p upregulation, and SIRT1 inhibition inhibited CGA-mediated anti-inflammatory and neuronal protective functions. There is a targeting correlation between MIR497HG, miR-29b-3p, and Sirt1. MIR497HG sponges miR-29b-3p to regulate SIRT1 expression in an indirect manner.
CONCLUSION
CGA upregulates MIR497HG to curb miR-29b-3p expression, hence initiating the SIRT1/NF-B signaling pathway and repressing OGD-elicited inflammation, oxidative stress, and neuron apoptosis.
背景
绿原酸（CGA）是一种具有抗氧化和抗炎特性的多酚化合物。CGA 已被证明可改善神经炎症。本研究旨在阐明 CGA 减轻神经炎症的确切机制。
方法
利用氧葡萄糖剥夺（OGD）处理 BV2 小胶质细胞和 HT-22 海马神经元，构建缺氧缺血再灌注模型。通过 ELISA 试剂盒测定小胶质细胞中炎症因子（IL-1、IL-6、TNF-、IL-4 和 IL-10）和氧化应激因子（MDA、SOD 和 GSH-PX）的水平。通过 CCK-8 测定神经元增殖，LDH 试剂盒测定神经元中 LDH 的释放。用荧光染料 DCF-DA 测定神经元中的 ROS 水平。通过 qRT-PCR 确定神经元和小胶质细胞中 MIR497HG、miR-29b-3p 和 SIRT1/NF-B 的相关性。通过 Western blot 测定小胶质细胞或神经元中炎症蛋白（COX2、iNOS）、氧化应激途径（Nrf2、HO-1）和凋亡相关蛋白（Bcl-2、Bax、caspase3、caspase8 和 caspase9）的表达。通过双荧光素酶测定和 RIP 测定确定 MIR497HG 和 miR-29b-3p 之间以及 miR-29b-3p 和 SIRT1 之间的相互作用。
结果
CGA 减轻了 OGD 介导的小胶质细胞炎症和氧化应激，并抑制了小胶质细胞介导的神经元凋亡。CGA 增加了 BV2 和 HT-22 细胞中 MIR497HG 和 SIRT1 的水平，抑制了 miR-29b-3p 的水平。MIR497HG 敲低、miR-29b-3p 上调和 SIRT1 抑制抑制了 CGA 介导的抗炎和神经元保护作用。MIR497HG、miR-29b-3p 和 Sirt1 之间存在靶向相关性。MIR497HG 海绵 miR-29b-3p 以间接方式调节 SIRT1 的表达。
结论
CGA 上调 MIR497HG 以抑制 miR-29b-3p 的表达，从而启动 SIRT1/NF-B 信号通路，抑制 OGD 诱导的炎症、氧化应激和神经元凋亡。
Zotero 插件