Clinical Medical College of Acupuncture-Moxibustion and Rehabilitation, Guangzhou University of Chinese Medicine, Guangzhou, China.
Department of Acupuncture and Moxibustion, The Affiliated TCM Hospital of Guangzhou Medical University, Guangzhou, China.
J Neuroinflammation. 2023 Feb 2;20(1):21. doi: 10.1186/s12974-023-02698-w.
The impairment in the autophagy-lysosomal pathway (ALP) and the activation of NLR family pyrin domain containing 3 (NLRP3) inflammasome represent two molecular events leading to neurodegeneration and neuroinflammation in Alzheimer's disease (AD), a devastating neurodegenerative disorder without a cure. Previously we demonstrated the cognitive-enhancing effect of a combined electroacupuncture (EA) therapy termed TNEA in a transgenic mouse model of AD, involving activation of transcription factor EB (TFEB), a master regulator of ALP. However, whether and how TNEA inhibits NLRP3 inflammasome via TFEB-mediated ALP in AD remains to be investigated.
5xFAD mice overexpressing amyloid-β (Aβ) were treated with TNEA or EA on its composing acupoints (GB13 and GV24). The changes in the signaling pathways regulating NLRP3 inflammasome, the association of NLRP3 inflammasome with ALP, and the roles of TFEB/TFE3 in mice brains were determined by immunoblots, immunohistochemistry and AAV-mediated knockdown assays.
TNEA inhibits the activation of NLRP3 inflammasome and the release of active interleukin 1β (IL1B) in the hippocampi of 5xFAD mice. Mechanistically, TNEA promoted the autophagic degradation of inflammasome components via activating both TFEB and TFE3 by modulating kinases including AMPK and AKT. The composing acupoints in TNEA showed synergistic effects on regulating these molecular events and memory improvement.
Our findings suggest that TNEA attenuates AD-associated memory impairment via promoting TFEB/TFE3-mediated autophagic clearance of Aβ and NLRP3 inflammasome, and partially reveal the molecular basis of combined acupoints therapy originated from ancient wisdom.
自噬溶酶体途径（ALP）的损伤和 NLR 家族富含 pyrin 域蛋白 3（NLRP3）炎性小体的激活是导致阿尔茨海默病（AD）神经退行性变和神经炎症的两个分子事件，AD 是一种破坏性的神经退行性疾病，目前尚无治愈方法。先前我们证明了一种名为 TNEA 的联合电针（EA）疗法在 AD 的转基因小鼠模型中的认知增强作用，该作用涉及转录因子 EB（TFEB）的激活，TFEB 是 ALP 的主要调节剂。然而，TNEA 是否以及如何通过 TFEB 介导的 ALP 抑制 AD 中的 NLRP3 炎性小体仍有待研究。
过表达淀粉样蛋白-β（Aβ）的 5xFAD 小鼠接受 TNEA 或其组成穴位（GB13 和 GV24）的 EA 治疗。通过免疫印迹、免疫组织化学和 AAV 介导的敲低测定来确定调节 NLRP3 炎性小体的信号通路变化、NLRP3 炎性小体与 ALP 的关联以及 TFEB/TFE3 在小鼠大脑中的作用。
TNEA 抑制 5xFAD 小鼠海马中 NLRP3 炎性小体的激活和活性白细胞介素 1β（IL1B）的释放。在机制上，TNEA 通过调节 AMPK 和 AKT 等激酶，激活 TFEB 和 TFE3，促进炎性小体成分的自噬降解。TNEA 的组成穴位在调节这些分子事件和改善记忆方面具有协同作用。
我们的研究结果表明，TNEA 通过促进 TFEB/TFE3 介导的 Aβ 和 NLRP3 炎性小体的自噬清除来减轻 AD 相关的记忆障碍，部分揭示了源自古代智慧的联合穴位疗法的分子基础。
