Abstract
OBJECTIVE:To assess the effect and mechanism of Sanhua Tang(三化汤,SHT)in treating ischemic stroke(IS)through the"Kaitong Xuanfu"theory by using network pharmacology and animal experiments.METHODS:The active ingredients and targets of SHT and IS were screened by public databases such as Traditional Chinese Medicine systems pharmacology,GeneCards,and online mendelian inheritance in man.Visual network topographies were constructed using R,Cytoscape 3.6.0,AutoDockTools,a user-sponsored molecular visualization system on an open-source foundation,and other software to analyze the correlation between targets and active ingredients.The middle cerebral artery occlusion(MCAO)model was established by operation.Animals were divided into the Sham group,MCAO group(M group),aloe-emodin(AE)group(MCAO rats treated with aloe-emodin),SHT at low dosage(SL group)(MCAO rats treated with SL),SHT at medium dosage(SM group),and SHT at high dosage(SH group).2,3,5-triphenyl tetrazolium chloride staining was used to detect the volume of cerebral infarction;Nissl staining was used to observe the morphology of neuronal cells;transmission electron microscopy was used to observe the integrity of the blood-brain barrier(BBB);enzyme-linked immunosorbent assay was used to detect the content of interleukin-6(IL-6),IL-10,tumor necrosis factorα(TNF-α)in serum.Western blot was used to detect the expression of vascular endothelial growth factor A(VEGFA)protein in the cerebral ischemic penumbra.RESULTS:Using network pharmacology and molecular docking validation,four active ingredients(lignan,naringenin,aloe-rhodopsin,and β-sitosterol),seven target proteins(protein kinase b 1,IL-6,TNF,VEGFA,TP53,jun proto-oncogene,and cysteinyl aspartate specific proteinase 3),and inflammatory signaling pathways were identified.Animal experiments showed that the SH and AE groups had fewer neurological deficits,reduced brain infarct volumes,decreased serum inflammatory factor levels,increased expression of VEGFA protein,and less structural damage to neurons and BBB.CONCLUSION:The present study found that the therapeutic mechanism of SHT against IS may be related to the inhibition of BBB inflammatory damage,which is also the mechanism of"Kaitong Xuanfu."The high-dose group of SHT was relatively effective in regulating inflammatory factors,improving BBB permeability,and protecting neuronal cells from damage.
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