HAGLR 通过从 ATAT1 上隔离 miR-182-5p 并激活 NLRP3 炎性小体，加重神经病理性疼痛，并促进脂多糖处理的 SH-SY5Y 细胞的炎症反应和细胞凋亡。
HAGLR aggravates neuropathic pain and promotes inflammatory response and apoptosis of lipopolysaccharide-treated SH-SY5Y cells by sequestering miR-182-5p from ATAT1 and activating NLRP3 inflammasome.
机构信息
Department of Anesthesiology, Second Affiliated Hospital of Soochow University, Suzhou, 215000, Jiangsu, China; Department of Pain Medical Center, Lianyungang Second People's Hospital, Lianyungang, 222000, Jiangsu, China.
Department of Anaesthesia, Affiliated Hospital of Xuzhou Medical University, Xuzhou, 221006, Jiangsu, China.
出版信息
Neurochem Int. 2021 May;145:105001. doi: 10.1016/j.neuint.2021.105001. Epub 2021 Feb 21.
BACKGROUND
Chronic neuropathic pain is characterized by neuroinflammation. Previously, long noncoding RNA (lncRNA) HAGLR was reported to regulate the inflammatory response of SH-SY5Y cells. However, neither the specific function nor the potential mechanism of HAGLR in neuropathic pain has been explored.
AIM OF THE STUDY
Our study is aimed to figure out the role of HAGLR in neuropathic pain.
METHODS
SH-SY5Y cells were treated with lipopolysaccharide (LPS) to mimic neuron injury in vitro. The chronic constriction injury (CCI) rat models were established by ligation of sciatic nerve to mimic neuropathic pain in vivo. Behavioral assessment assays were performed to determine the effects of HAGLR on hypersensitivity in neuropathic pain. Enzyme-linked immunosorbent assay kits were used for detection of inflammatory cytokines. Flow cytometry analysis and Western blot were applied to detect apoptosis.
RESULTS
HAGLR displayed high levels in spinal cords of CCI rats and in LPS treated SH-SY5Y cells. Knockdown of HAGLR inhibited inflammation and neuron apoptosis of LPS treated SH-SY5Y cells. Mechanistically, HAGLR bound with miR-182-5p in SH-SY5Y cells. ATAT1 served as a target of miR-182-5p. HAGLR activated the NLRP3 inflammasome by ATAT1. Rescue assays demonstrated that overexpression of ATAT1 or NLRP3 reversed the suppressive effects of HAGLR silencing on apoptosis and inflammatory response in SH-SY5Y cells and in spinal cords of CCI rats. The inhibitory effects of silenced HAGLR on hypersensitivity in neuropathic pain were also rescued by ATAT1 or NLRP3.
CONCLUSIONS
HAGLR aggravates neuropathic pain by sequestering miR-182-5p from ATAT1 and activating NLRP3 inflammasome, which may provide a potential therapeutic target for neuropathic pain treatment.
背景
慢性神经性疼痛的特征是神经炎症。先前有研究报道长链非编码 RNA（lncRNA）HAGLR 可调节 SH-SY5Y 细胞的炎症反应。然而，HAGLR 在神经性疼痛中的具体功能及其潜在机制尚未得到探索。
目的
本研究旨在探讨 HAGLR 在神经性疼痛中的作用。
方法
用脂多糖（LPS）处理 SH-SY5Y 细胞以模拟体外神经元损伤。通过结扎坐骨神经建立慢性缩窄性损伤（CCI）大鼠模型以模拟体内神经性疼痛。行为评估实验用于确定 HAGLR 对神经性疼痛过敏的影响。酶联免疫吸附试剂盒用于检测炎症细胞因子。流式细胞术分析和 Western blot 用于检测细胞凋亡。
结果
HAGLR 在 CCI 大鼠脊髓和 LPS 处理的 SH-SY5Y 细胞中表达水平较高。敲低 HAGLR 抑制了 LPS 处理的 SH-SY5Y 细胞的炎症和神经元凋亡。机制上，HAGLR 在 SH-SY5Y 细胞中与 miR-182-5p 结合。ATAT1 是 miR-182-5p 的靶基因。HAGLR 通过 ATAT1 激活 NLRP3 炎性小体。挽救实验表明，过表达 ATAT1 或 NLRP3 逆转了 HAGLR 沉默对 LPS 处理的 SH-SY5Y 细胞和 CCI 大鼠脊髓中细胞凋亡和炎症反应的抑制作用。沉默 HAGLR 对神经性疼痛过敏的抑制作用也被 ATAT1 或 NLRP3 挽救。
结论
HAGLR 通过从 ATAT1 上隔离 miR-182-5p 并激活 NLRP3 炎性小体加重神经性疼痛，这可能为神经性疼痛治疗提供一个潜在的治疗靶点。
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