Research Paper Advance Articles

Dl-3-n-butylphthalide inhibits neuroinflammation by stimulating foxp3 and Ki-67 in an ischemic stroke model

Xi Liu1,2, *, , Runzhe Liu1,2, *, , Dongxu Fu3, *, , Hao Wu1,2, , Xin Zhao1,2, , Yi Sun1,2, , Meng Wang3, , Xiaoping Pu1,2, ,

  • 1 National Key Research Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing 100191, P.R. China
  • 2 Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Sciences, Peking University, Beijing 100191, P.R. China
  • 3 CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049, P.R. China
* Equal contribution

Received: May 2, 2020       Accepted: November 20, 2020       Published: January 10, 2021      

https://doi.org/10.18632/aging.202338
How to Cite

Copyright: © 2021 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Dl-3-n-butylphthalide (NBP) has been widely used to treat ischemic stroke in China. To investigate the mechanisms underlying NBP activity, we established a permanent middle cerebral artery occlusion (pMCAO) rat model and injected the rats with 4 mg/kg/d NBP for nine days. We then assessed neuroinflammation, neovascularization and nerve regeneration within the brain. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry imaging (MALDI-TOF MSI) was used to determine the phospholipid distribution, while laser ablation-inductively coupled plasma mass spectrometry imaging (LA-ICP MSI) was used to measure Foxp3, Ki-67 and pCREB levels in the brain. Immunohistochemistry was used to investigate the expression of NLR family pyrin domain containing 3 (NLRP3) and its inflammatory products, caspase-1 and interleukin-1β, in brain tissues. NBP attenuated ischemic damage and ameliorated neurological deficits in rats with pMCAO. In the ischemic brain region, NBP reduced phosphatidylethanolamine (18:0), NLRP3, caspase-1 and interleukin-1β levels, but increased levels of Foxp3, Ki-67, pCREB and several phospholipids. In molecular docking analyses, NBP bound to NLRP3, interleukin-1β, caspase-1, Foxp3, and Ki-67. These results demonstrate that NBP reduces neuroinflammation in brain tissues and promotes nerve and blood vessel regeneration, thus protecting neuromorphology and function.

Abbreviations

NBP: Dl-3-n-butylphthalide; UK: Urinary kallidinogenase; pMCAO: Permanent middle cerebral artery occlusion; MALDI-TOF MSI: Matrix-assisted laser desorption ionization time-of-flight mass spectrometry imaging; LA-ICP MSI: Laser ablation-inductively coupled plasma mass spectrometry imaging; mNSS: Modified neurological severity score; Treg: Regulatory T.