Abstract

Background: Cerebral ischemia-reperfusion can cause structural changes in brain tissue and disrupt physiological functions. Naoluotong granules (NLT) exhibit significant neuroprotective effects on brain nerve cells and are widely utilized in treating cerebral ischemia-reperfusion injury (CIRI). However, the underlying molecular mechanisms remain elusive. Methods: The CIRI model was established in vivo , with rats assigned to four groups. 2,3,5-triphenyltetrazolium chloride (TTC), hematoxylin and eosin (H&E), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) were used to evaluate the effects of NLT and AST-IV on neurological function and pathological injury after cerebral ischemia-reperfusion. immunofluorescence, real-time PCR, and Western-blot assays were performed to explore the effects of NLT and AST-IV on the RIP1/NIK/IKKα/NF-κB pathway. The micro-enzyme immunoassay method was applied to examine the expression levels of various factors in each group. To verify the neuroprotective effects of NLT and AST-IV in vitro , an oxygen-glucose deprivation/reoxygenation (OGD/R) PC12 cell model was established and treated with the drug-containing serum of NLT and AST-IV. Results: Necroptosis plays a significant role in the onset and progression of ischemia-reperfusion-induced neural injury. In vivo studies have shown that both NLT and AST-IV improve neurological function scores, reduce infarct volume, and enhance pathological features following MCAO/R. They significantly inhibited the expression of the RIP1/NIK/IKKα/NF-κB pathway, thereby reducing neuronal cell necroptosis. In vitro studies also showed that NLT and AST-IV reduced OGD/R-induced necroptosis. Conclusions: This study confirms that the significant neuroprotective effects of NLT and AST-IV in alleviating CIRI are related to inhibiting necrosis by suppressing the RIP1/NIK/IKKα/NF-κB pathway.