2. Academic Validation
  3. Probenecid relieves LPS-induced neuroinflammation by modulating the NLRP1 inflammasome pathway in BV2 cells

Probenecid relieves LPS-induced neuroinflammation by modulating the NLRP1 inflammasome pathway in BV2 cells

  • Sci Rep. 2025 Oct 8;15(1):35192. doi: 10.1038/s41598-025-19015-5.
Xiaoxuan Wang # 1,  Haozhe Tian # 2 3,  Shifei Chen 1,  Liangliang Ying 1,  Juan Zheng 2 3,  Yaya Song 1,  Jin Xi 3,  Yuxin Zhang 4 5,  Dong Wang 6
Affiliations

Affiliations

  • 1 Clinical Laboratory, Second People's Hospital of Bengbu City, Bengbu, 233000, China.
  • 2 Key Laboratory of Cancer Research and Clinical Laboratory Diagnosis, Bengbu Medical University, Bengbu, 233030, China.
  • 3 Anhui Key Laboratory of Tissue Transplantation, Bengbu Medical University, Bengbu, 233030, China.
  • 4 Key Laboratory of Cancer Research and Clinical Laboratory Diagnosis, Bengbu Medical University, Bengbu, 233030, China. zhangyuxin@bbmc.edu.cn.
  • 5 Anhui Key Laboratory of Tissue Transplantation, Bengbu Medical University, Bengbu, 233030, China. zhangyuxin@bbmc.edu.cn.
  • 6 Clinical Laboratory, Second People's Hospital of Bengbu City, Bengbu, 233000, China. whxdong@126.com.
  • # Contributed equally.
PMID: 41062518 DOI: 10.1038/s41598-025-19015-5
Abstract

Inflammation is a key factor in neurodegenerative diseases (NDs) development, such as Alzheimer's disease (AD) and Parkinson's disease (PD). Microglial cells maintain immune microenvironment homeostasis in the central nervous system (CNS) and are actively involved in neuroinflammation. Therefore, inhibiting microglia-mediated neuroinflammation may provide therapeutic benefits in treating NDs. Probenecid (Prob), a sulfonamide derivative, has demonstrated neuroprotective and anti-inflammatory properties. This study investigated the anti-inflammatory and antioxidative properties of Prob in a neuroinflammatory model which was established by stimulating BV2 cells with lipopolysaccharide (LPS). The oxidative stress was assessed by measuring the nitric oxide (NO), Reactive Oxygen Species (ROS), superoxide dismutase (SOD), and malondialdehyde (MDA) levels. Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were quantified using enzyme-linked immunosorbent assay (ELISA) and RT-qPCR. The wound-healing assay and phagocytosis assay were employed to examine the migration and phagocytosis capacity of BV2 cells, respectively. The mRNA expression of CD16, CD206, iNOS, and Arg-1 was measured using RT-qPCR, while Western blot was used to analyze inflammasome-associated protein levels. The results indicated that Prob suppressed LPS-induced microglial activation and reduced TNF-α and IL-1β production. Moreover, Prob attenuated the migration and phagocytosis capacities of BV2 cells and inhibited oxidative stress. Prob regulated the immune microenvironment, as shown by reduced M1 markers (iNOS and CD16) and increased M2 markers (Arg-1 and CD206). However, the NLRP1 inflammasome activator (muramyl dipeptide, MDP) reversed the effects of Prob. In conclusion, Prob could alleviate LPS-induced inflammation by modulating the NLRP1 inflammasome pathway.

Keywords

Immune microenvironment; Inflammasomes; Neuroinflammation; Oxidative stress; Probenecid.

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