OBJECTIVE:To observe regulatory effect of Naoluoxintong formula(脑络欣通方,NLXT)and its split prescriptions on vascular regeneration of rats suffering from cerebral ischemia-reperfusion(IR)syndrome of Qi deficiency wi...OBJECTIVE:To observe regulatory effect of Naoluoxintong formula(脑络欣通方,NLXT)and its split prescriptions on vascular regeneration of rats suffering from cerebral ischemia-reperfusion(IR)syndrome of Qi deficiency with blood stasis(QDBS).METHODS:NLXT is the representative prescription of Yiqi Huoxue Tongluo decoction(益气活血通络方),and NLXT is divided into Yiqi herbs(益气药)and Huoxue Tongluo herbs(活血通络药)according to their efficacies.One hundred and eight specific-pathogen-free,clean-grade,Sprague-Dawley male rats were selected to prepare the classical rat model with QDBS due to middle artery ischemia-reperfusion using the multi-factor compound simulation approach.The animals were classified into sham operation(S),model(M),Nimodping(NMDP),NLXT,YQ and HXTL groups,each having 18 rats.Cerebral ischemia was reperfused after 2 h,and 24 h later,they were administered traditional Chinese medicine treatment for 14 d twice a day.Angiogenesis changes after NLXT administration to middle cerebral artery occlusion-reperfusion(MCAO/R)rats with QDBS were analyzed using the neurological deficit score and hematoxylin-eosin staining.Cerebral infarct area by 2,3,5-Triphenyltetrazolium chloride was detected,and the ultrastructure of the blood vessel in the ischemic frontoparietal cortex was observed by transmission electron microscopy.Angiopoietin 1(Ang1),angiopoietin 2(Ang2),vascular endothelial growth factor A(VEGFA),vascular endothelial growth factor receptor 2(VEGFR2),platelet endothelial cell adhesion molecule-1(CD31),angiopoietin receptor 2(Tie2),and P38 mitogen-activated protein kinase(MAPK)protein levels in the frontal and parietal cortex were quantified by immunofluorescence,reverse transcription-polymerase chain reaction,and Western blotting assays.RESULTS:Relative to the S group,VEGFA and VEGFR2 levels in the frontal and parietal cortex of group M were increased,and Ang1,Ang2,Tie2,CD31,and p38 MAPK levels remarkably increased(P<0.05);cerebral infarct area was significant and pathological morphology and ultrastructure damage was obvious.Relative to the group M,VEGFA,VEGFR2,CD31,Ang1,Ang2,and Tie2 expression of group NLXT and NMDP remarkably elevated(P<0.05)and infarct focus,pathological morphology and ultrastructure were significantly improved;VEGFA and VEGFR2 levels in the groups YQ and HXTL increased,and Ang1,Ang2,CD31,and Tie2 levels remarkably increased(P<0.05);p38 MAPK levels in the three treatment groups decreased(P<0.05).Relative to the group NLXT,the expression levels of p38 MAPK in group YQ and group HXTL were significantly increased,and the expression levels of other indicators were significantly decreased(P<0.05).CONCLUSION:NLXT can promote the angiogenesis of the rat model of MCAO/R with QDBS by activating VEGFA and inhibiting P38 MAPK,and the effect is better than that of split prescription groups.展开更多
基金Second Round Construction Project of state administration of Traditional Chinese Medicine Academic School Inheritance Studio[Letter of Human Teaching(2019)No.62 from State Administration of Traditional Chinese Medicine],the 13th Batch of"Innovation Team for Prevention and Treatment of Encephalopathy by Combination of Acupuncture and Medicine"Project of 115 Industry Innovation Team in Anhui Province[Anhui Talent Office(2020)No.4]Anhui Provincial Natural Science Foundation of China,studied the effects of Yiqi Huoxue Tongluo Method on Vascular Regeneration in Rats with Cerebral Ischemia and its Mechanism Based on Hypoxia-Inducible Factor 1Α/Vascular Endothelial Growth Factor Signaling Pathway(KJ2022A0547)。
文摘OBJECTIVE:To observe regulatory effect of Naoluoxintong formula(脑络欣通方,NLXT)and its split prescriptions on vascular regeneration of rats suffering from cerebral ischemia-reperfusion(IR)syndrome of Qi deficiency with blood stasis(QDBS).METHODS:NLXT is the representative prescription of Yiqi Huoxue Tongluo decoction(益气活血通络方),and NLXT is divided into Yiqi herbs(益气药)and Huoxue Tongluo herbs(活血通络药)according to their efficacies.One hundred and eight specific-pathogen-free,clean-grade,Sprague-Dawley male rats were selected to prepare the classical rat model with QDBS due to middle artery ischemia-reperfusion using the multi-factor compound simulation approach.The animals were classified into sham operation(S),model(M),Nimodping(NMDP),NLXT,YQ and HXTL groups,each having 18 rats.Cerebral ischemia was reperfused after 2 h,and 24 h later,they were administered traditional Chinese medicine treatment for 14 d twice a day.Angiogenesis changes after NLXT administration to middle cerebral artery occlusion-reperfusion(MCAO/R)rats with QDBS were analyzed using the neurological deficit score and hematoxylin-eosin staining.Cerebral infarct area by 2,3,5-Triphenyltetrazolium chloride was detected,and the ultrastructure of the blood vessel in the ischemic frontoparietal cortex was observed by transmission electron microscopy.Angiopoietin 1(Ang1),angiopoietin 2(Ang2),vascular endothelial growth factor A(VEGFA),vascular endothelial growth factor receptor 2(VEGFR2),platelet endothelial cell adhesion molecule-1(CD31),angiopoietin receptor 2(Tie2),and P38 mitogen-activated protein kinase(MAPK)protein levels in the frontal and parietal cortex were quantified by immunofluorescence,reverse transcription-polymerase chain reaction,and Western blotting assays.RESULTS:Relative to the S group,VEGFA and VEGFR2 levels in the frontal and parietal cortex of group M were increased,and Ang1,Ang2,Tie2,CD31,and p38 MAPK levels remarkably increased(P<0.05);cerebral infarct area was significant and pathological morphology and ultrastructure damage was obvious.Relative to the group M,VEGFA,VEGFR2,CD31,Ang1,Ang2,and Tie2 expression of group NLXT and NMDP remarkably elevated(P<0.05)and infarct focus,pathological morphology and ultrastructure were significantly improved;VEGFA and VEGFR2 levels in the groups YQ and HXTL increased,and Ang1,Ang2,CD31,and Tie2 levels remarkably increased(P<0.05);p38 MAPK levels in the three treatment groups decreased(P<0.05).Relative to the group NLXT,the expression levels of p38 MAPK in group YQ and group HXTL were significantly increased,and the expression levels of other indicators were significantly decreased(P<0.05).CONCLUSION:NLXT can promote the angiogenesis of the rat model of MCAO/R with QDBS by activating VEGFA and inhibiting P38 MAPK,and the effect is better than that of split prescription groups.
文摘目的采用网络药理学方法探究益气活血方脑络欣通治疗缺血性脑卒中(ischemic stroke,IS)的作用机制,并通过分子对接技术进行初步验证。方法通过TCMSP、ETCM、化学专业数据库及文献筛选脑络欣通活性成分及作用靶点,通过OMIM、DisGeNET、GeneCards以及Drugbank数据库挖掘IS相关靶点;将交集靶点导入STRING平台和Cytoscape 3.8.2软件构建PPI网络并通过拓扑分析获得核心靶点,同时构建“中药—活性成分—核心靶点—疾病”可视化网络图;运用R软件进行GO功能及KEGG通路富集分析。最后借助Auto Dock Tools软件以及PyMOL软件进行分子对接和可视化。结果脑络欣通的潜在活性成分78个,治疗IS的潜在作用靶点200个,核心靶点包括STAT3、JUN、MAPK1、TP53等;作用较突出的活性成分包括槲皮素、β-谷甾醇、山柰酚、木犀草素等。进一步的生物功能分析获得150条信号通路,包括PI3K-AKT信号通路、HIF-1信号通路、FoxO信号通路等关键通路。分子对接结果表明关键活性成分与核心靶点之间存在分子结合位点,并有较强的结合活性。结论益气活血方脑络欣通可能是通过多成分、多靶点协同作用于氧化应激、炎症反应、细胞凋亡、自噬等生理病理环节,从而发挥治疗IS的作用。
