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

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.

L-3-n-butylphthalide promotes restoration after an experimental animal model of intracerebral hemorrhage

“…The preclinical studies revealed that NBP exerts neuroprotective effects in ischemic stroke both in vivo and in vitro , partially by promoting angiogenesis 30 , increasing BDNF expression 12 , promoting dendrite development 31 , protecting mitochondrial function 9 , inhibiting neuroinflammation 32 , and improving brain metabolism 33 . The clinical trials also demonstrated that NBP exerts anti-ischemic effects in patients.…”

Challenges and Improvements of Novel Therapies for Ischemic Stroke

“…Till 1995, NBP was found to be useful for ischemic stroke and later experiments showed that both injecting NBP 3 h before or 2 h after surgery had improvement on infarction volume and motor functional recovery, and clinical studies showed that patients using NBP alone had better neurological function than control groups; however, there was no significant difference for the efficacy between NBP and tPA, so NBP had been used for stroke treatment in China ( Xu et al, 2019 ). Anti-inflammation was firstly found in the NBP-treated stroke model; NBP enhanced the proliferation of microglia and promoted the microglia polarization into M2 phenotype, an anti-inflammation phenotype, by activating the calcium/calmodulin-dependent protein kinase β (CaMKKβ)-AMPK pathway or PPARγ nuclear translocation; NBP also decreased the infiltration of endothelial cells by downregulating intercellular adhesion molecule 1 (ICAM-1) and protease-activated receptor-1 (PAR-1) ( Zeng et al, 2020 ; Liu et al, 2021b ). In addition, NBP played a role in promoting neuroplasticity, neurite outgrowth, and inhibiting neuron apoptosis, and the NgR, Mogo-A, and caspase 3 were decreased and growth factors such as BDNF, NGF, and neurite branches were increased ( Zhang et al, 2021 ).…”

Rescue of Mitochondrial SIRT3 Ameliorates Ischemia-like Injury in Human Endothelial Cells

“…NBP has multiple pharmacologic effects in ischemic stroke, not only of endothelial protection but also of neuroprotection, anti-oxidation and anti-platelet aggregation [ 2 , 7 ]. In order to verify our hypothesis, we further compared the protective role of OHNBP and NBP in neuroprotection, anti-oxidation and anti-platelet aggregation.…”

“…Currently, thrombolysis using intravenous recombinant tissue plasminogen activator (rtPA) is the only therapy approved by the FDA for ischemic stroke, while it can only be delivered to a small fraction of patients due to the narrow treatment window [ 1 ]. Other treatments including antiplatelet (such as clopidogrel, aspirin), neuroprotection (such as NBP) and antioxidant agents (such as edaravone) are also implemented in clinical practice, which provide limited benefits to patients [ 2 , 3 , 4 , 5 , 6 , 7 ]. A better understanding of stroke pathology is important to identify innovative targets to broaden our therapeutic options in this scenario.…”