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...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);enzymelinked 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,andβ-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.展开更多
OBJECTIVE:To explore the protective mechanisms of the Traditional Chinese Medicine Bushenhuoxue(BSHX)in a rat model of vascular dementia(VD).METHODS:A rat model of VD was developed using bilateral common carotid arter...OBJECTIVE:To explore the protective mechanisms of the Traditional Chinese Medicine Bushenhuoxue(BSHX)in a rat model of vascular dementia(VD).METHODS:A rat model of VD was developed using bilateral common carotid artery occlusion(BCCAO).Rats were administered BSHX(10.14 or 5.07 g/kg),nimodipine(11.06 mg/kg;positive control),or saline(control)by gavage daily for 30 d post-surgery.Learning and memory abilities were assessed using the Morris water maze.Morphological changes in the hippocampus were observed using light microscopy(hematoxylin and eosin staining)and transmission electron microscopy(TEM).The m RNA and protein expression levels of brain-derived neurotrophic factor(BDNF),tyrosine receptor kinase B(Trk B),phosphatidyl inositol 3-kinase(PI3 K),serine/threonine kinase(AKT),and c AMP response element binding protein(CREB)were measured by real-time polymerase chain reaction(RT-PCR)and Western blot,respectively.RESULTS:Compared with the sham group,rats with BCCAO exhibited impaired learning and memory abilities(Morris water maze)and showed abnormalities in neuronal morphology(light microscopy)and ultrastructure(TEM)in the hippocampus.They also had decreased m RNA and protein expressions of BDNF,Trk B,PI3 K,AKT,and CREB in hippocampal tissue(all P<0.05).In rats with BCCAO,administration of BSHX attenuated deficits in learning and memory,improved the morphology and ultrastructure of hippocampal neurons,and enhanced m RNA and protein expression levels of BDNF,Trk B,PI3 K,AKT,and CREB(all P<0.05).CONCLUSION:BSHX may protect hippocampal neurons and improve learning and memory abilities,at least in part via the activation of BDNF/Trk B/PI3 K/AKT/CREB signaling.展开更多
基金Hebei Province Natural Science:Based on The Xuan Fu Theory to Explore Sanhua Tang on Ischemic Stroke Blood-brain Barrier Protection Mechanism Research(No.H2022423327)Science and Technology Program of Hebei Provincial Administration of Traditional Chinese Medicine:based on The Changes in Gut Microbiota,Explore Sanhua Tang From The Perspective of"Opening the Xuan Fu"Protective Phlegm Heat Relieving Excess Syndrome of Ischemic Stroke Mechanism of Action of the Blood-brain Barrier(No.2022090)+3 种基金Hebei Key Laboratory of Chinese Medicine Research on Cardio-cerebrovascular Disease in 2021 Project:Based on the Gut-brain Axis,Explore Sanhua Tang's Treatment of Ischemic Stroke Syndrome(Phlegm Heat Relieving Empirical Model)(No.2021201)the Innovation Funding Program for Doctoral Students in Hebei Province:the Mechanism Research of Sanhua Tang Regulating Gut Microbiota,Improving the Blood-brain Barrier Damage of Ischemic Stroke(No.CXZZBS2022095)National College Students'Innovation and Entrepreneurship Training Program:Based on the Xuan Fu Theory to Explore Sanhua Tang on Ischemic stroke Blood-brain Barrier Protection Mechanism Research(No.202114432005)Special Project on the Cultivation of Scientific and Technological Innovation Ability of College and Middle School Students in Hebei Province:Mechanism Research of Bushen Huoxue Fang on Synaptic Plasticity in the Hippocampal Nerve After Vascular Dementia through Phosphatidylinositide 3-Kinase/Protein Kinase b/Mammalian Target of Rapamycin Signaling Pathways(22E50142D)。
文摘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);enzymelinked 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,andβ-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.
基金Supported by Hebei Province Natural Science Fund(Protection Mechanism Research of Bushenhuoxue on Hippocampal Nerve in Rats with Vascular Dementia Based on BDNF/Trk B Signaling Pathway,No.H2015423057)Hebei Provincial Department of Education Science and Technology Research Key Funding Project(Study on the Mechanism of Regulating the Autophagy and Apoptosis of Hippocampal Neurons in VD Rats by Bushenhuoxue Prescription,No.ZD2018009)Research Capacity Enhancement Project of Hebei University of Chinese Medicine(Improvement of Hippocampal Synaptic Remodeling in Rats with Vascular Dementia by Regulating BDNF/Trk B Signaling Pathway Based on the"renal essence"theory,No.2019-11).
文摘OBJECTIVE:To explore the protective mechanisms of the Traditional Chinese Medicine Bushenhuoxue(BSHX)in a rat model of vascular dementia(VD).METHODS:A rat model of VD was developed using bilateral common carotid artery occlusion(BCCAO).Rats were administered BSHX(10.14 or 5.07 g/kg),nimodipine(11.06 mg/kg;positive control),or saline(control)by gavage daily for 30 d post-surgery.Learning and memory abilities were assessed using the Morris water maze.Morphological changes in the hippocampus were observed using light microscopy(hematoxylin and eosin staining)and transmission electron microscopy(TEM).The m RNA and protein expression levels of brain-derived neurotrophic factor(BDNF),tyrosine receptor kinase B(Trk B),phosphatidyl inositol 3-kinase(PI3 K),serine/threonine kinase(AKT),and c AMP response element binding protein(CREB)were measured by real-time polymerase chain reaction(RT-PCR)and Western blot,respectively.RESULTS:Compared with the sham group,rats with BCCAO exhibited impaired learning and memory abilities(Morris water maze)and showed abnormalities in neuronal morphology(light microscopy)and ultrastructure(TEM)in the hippocampus.They also had decreased m RNA and protein expressions of BDNF,Trk B,PI3 K,AKT,and CREB in hippocampal tissue(all P<0.05).In rats with BCCAO,administration of BSHX attenuated deficits in learning and memory,improved the morphology and ultrastructure of hippocampal neurons,and enhanced m RNA and protein expression levels of BDNF,Trk B,PI3 K,AKT,and CREB(all P<0.05).CONCLUSION:BSHX may protect hippocampal neurons and improve learning and memory abilities,at least in part via the activation of BDNF/Trk B/PI3 K/AKT/CREB signaling.
