Mou Yu-Jie, Li Feng-Min, Lou Jun-Tong, Tu Hai-Yue, Zhu Yi, Sheng Rui, Zhang Zhong-Ling, Zhao Yu-Zheng, Ji Fu-Hai, Wu Jun-Chao, Qin Zheng-Hong
Department of Anesthesiology, the First Affiliated Hospital of Soochow University, Suzhou, 215006, China.
Department of Pharmacology and Laboratory of Aging and Nervous Diseases, Jiangsu Key Laboratory of Neuropsychiatric Diseases, College of Pharmaceutical Sciences, Soochow University, Suzhou, 215123, China.
Acta Pharmacol Sin. 2025 Aug 11. doi: 10.1038/s41401-025-01638-z.
Reduced nicotinamide adenine dinucleotide phosphate (NADPH) is an important coenzyme involved in cellular biosynthetic and redox metabolism. It has been recognized for its role in regulating neuroinflammation through coordinating redox reactions. Whether there are new actions other than redox regulation remain unclear. In this study we investigated a novel mechanism by which NADPH regulated microglia-mediated neuroinflammation. We showed that NADPH application significantly alleviated NLRP3 inflammasome activation in microglia and exerted neuroprotective effects both in vitro and in vivo neuroinflammation models. With P2X7R knockdown microglial cells and P2X7R mice, we demonstrated that P2X7R was a crucial mediator of the anti-inflammatory effects for the supplemented NADPH. We conducted whole-cell recording from murine microglial cell line BV2 cells, and found that application of ATP (1 mM) elicited an inward current, which was reduced by co-application of P2X7R antagonist A-438079 (20 μM) or NADPH (1 mM). By performing a drug affinity responsive targets stability (DARTS) assay, we revealed that NADPH (not NADP or NADH), like the P2X7R agonist ATP, bound to the extracellular domain of P2X7R, leading to the suppression of ATP-induced P2X7R activation. Our research provides the first evidence of NADPH as an endogenous inhibitor of P2X7R in modulation of microglia-mediated neuroinflammation. This study expands the biological functions of NADPH and offers a novel target for NADPH-based therapies in neuroimmune-related diseases.
还原型烟酰胺腺嘌呤二核苷酸磷酸（NADPH）是一种参与细胞生物合成和氧化还原代谢的重要辅酶。它通过协调氧化还原反应在调节神经炎症中的作用已得到认可。除了氧化还原调节外是否存在其他新作用仍不清楚。在本研究中，我们探究了NADPH调节小胶质细胞介导的神经炎症的新机制。我们发现，应用NADPH可显著减轻小胶质细胞中NLRP3炎性小体的激活，并在体外和体内神经炎症模型中发挥神经保护作用。利用P2X7R基因敲低的小胶质细胞和P2X7R基因敲除小鼠，我们证明P2X7R是补充NADPH抗炎作用的关键介质。我们对小鼠小胶质细胞系BV2细胞进行全细胞记录，发现应用ATP（1 mM）可引发内向电流，而联合应用P2X7R拮抗剂A-438079（20 μM）或NADPH（1 mM）可使其减小。通过进行药物亲和力响应靶点稳定性（DARTS）分析，我们发现NADPH（而非NADP或NADH）与P2X7R激动剂ATP一样，与P2X7R的胞外结构域结合，从而抑制ATP诱导的P2X7R激活。我们的研究首次证明NADPH作为P2X7R的内源性抑制剂在调节小胶质细胞介导的神经炎症中发挥作用。本研究扩展了NADPH的生物学功能，并为神经免疫相关疾病基于NADPH的治疗提供了新靶点。
