Impact of sanguinarine in protecting neuronal damage in rats with ischemic stroke by regulating the RIPK1/RIPK3/MLKL axis

CHEN Wei; JIANG Xuelin; HUANG Xulin

doi:10.3969/j.issn.1002-7386.2026.01.006

Abstract：Objective To investigate the impact of sanguinarine(SAG)on neuronal damage in rats with ischemic stroke(IS)by regulating the receptor interacting protein kinase 1/3(RIPK1/RIPK3)/mixed lineage kinase domain-like protein(MLKL)axis.Methods A total of 72 specific pathogen free(SPF)Sprague-Dawley(SD)rats were randomly divided into Sham group,Model group,low-dose,medium-dose and high-dose SAG groups,and positive control group(nimodipine).The modified middle cerebral artery occlusion(MCAO)was performed in rats to prepare the IS model.Loeffler neurological score was graded to evaluate the nerve function and damage degree of rats.The area of cerebral infarction was measured by 2,3,5-triphenyltetrazolium chloride(TTC)staining.The water content of rat brain tissue was measured by weighing.Hematoxylin and eosin(H&E)staining staining was applied to observe the pathological damage of rat hippocampus.Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)staining was applied to observe the apoptosis of neurons in rats.The levels of interleukin(IL)-6,tumor necrosis factor-α(TNF-α)and IL-β in rat hippocampus were detected by Enzyme-linked immunosorbent assay(ELISA).Lactate dehydrogenase(LDH),catalase(CAT),malondialdehyde(MDA)and reactive oxygen species(ROS)in rat hippocampus were measured by commercial kits.Western blot was applied to detect the protein expressions of caspase-8,RIPK1,RIPK3 and MLKL in rat hippocampus.Results Compared with sham group,apoptosis rate of nerve cells,water content of brain tissue,percentage of cerebral infarction area,levels of IL-6,IL-β,and TNF-α,LDH release rate,contents of MDA,and ROS,and protein levels of RIPK1,RIPK3,and p-MLKL/MLKL in the model group were significantly higher,while the neurological score,CAT activity and protein level of caspase-8 were significantly lower(P<0.05).Compared with the model group,the neurological score,CAT activity and protein level of caspase-8 in the low-dose,medium-dose and high-dose SAG groups and the positive control group were significantly higher,while the percentage of cerebral infarction area,water content of brain tissue,apoptosis rate of nerve cells,levels of IL-6,IL-β,and TNF-α,LDH release rate,contents of MDA and ROS,and the protein levels of RIPK1,RIPK3,and p-MLKL/MLKL were significantly lower,and the damage of hippocampus was significantly reduced(P<0.05).Conclusion SAG can inhibit oxidative stress and inflammatory reaction,reduce the apoptosis of nerve cells,and play a neuroprotective role on IS rats by inhibiting the RIPK1/RIPK3/MLKL axis.