Objective:To observe the changes of coagulation and fibrinolysis function in human umbilical cord vein endothelial cells (HUVEC) induced by recombinant human tumor necrosis factor-α (rhT-NF-α) and the effect of Naot...Objective:To observe the changes of coagulation and fibrinolysis function in human umbilical cord vein endothelial cells (HUVEC) induced by recombinant human tumor necrosis factor-α (rhT-NF-α) and the effect of Naotaifang extract (脑泰方, NTE) on it. Methods: Cultured HUVEC is randomly divided into six groups:Control group, NTE control group (only 2 g/L NTE), rhTNF-α group (100ug/ L rhTNF-α), and low-dosage, middle-dosage and high-dosage NTE group (100ug/L rhTNF-α and 0. 67 g/L, 2 g/L, 6 g/L NTE). The coagulation activity of frozen-dissolved HUVEC, von Willebrand factor (vWF) content in the conditioned medium and tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor (PAD activity were to be detected after 24 hrs. Results: Compared with the control group, PAI activity were enhanced, vWF release markedly increased in conditioned medium of TNF-α group (P<0. 01) and the frozen-dissolved HUVEC markedly shortens the rabbit plasma prothrombin time, and the above changes could be significantly inhibited by the 3 dosages of NTE (P<0. 05, P<0. 01). Conclusion:NTE is effective in inhibiting the coagulation activity of the HUVEC non-stimulated or stimulated by rhTNF-a to enhance the vWF release, and to adjust fibrinolytic function, and mainly to inhibit the PAI activity.展开更多
Objective:Our previous research showed that Naotaifang(a compound traditional Chinese herbal medicine)extract(NTE)has clinically beneficial effects on neurological improvement of patients with acute cerebral ischemia....Objective:Our previous research showed that Naotaifang(a compound traditional Chinese herbal medicine)extract(NTE)has clinically beneficial effects on neurological improvement of patients with acute cerebral ischemia.In this study,we investigated whether NTE protected acute brain injury in rats and whether its effects on ferroptosis could be linked to the dysfunction of glutathione peroxidase 4(GPX4)and iron metabolism.Methods:We established an acute brain injury model of middle cerebral artery occlusion(MCAO)in rats,in which we could observe the accumulation of iron in neurons,as detected by Perl’s staining.Using assay kits,we measured expression levels of ferroptosis biomarkers,such as iron,glutathione(GSH),reactive oxygen species(ROS)and malonaldehyde(MDA);further the expression levels of transferrin receptor1(TFR1),divalent metal transporter 1(DMT1),solute carrier family 7 member 11(SLC7 A11)and GPX4 were determined using immunohistochemical analysis,real-time quantitative polymerase chain reaction and Western blot assays.Results:We found that treatment with NTE reduced the expression levels of TFR1 and DMT1,reduced ROS,MDA and iron accumulation and reduced neurobehavioral scores,relative to untreated MCAO rats.Treatment with NTE increased the expression levels of SLC7 A11,GPX4 and GSH,and the number of Nissl bodies in the MCAO rats.Conclusion:Taken together,our data suggest that acute cerebral ischemia induces neuronal ferroptosis and the effects of treating MCAO rats with NTE involved inhibition of ferroptosis through the TFR1/DMT1 and SCL7 A11/GPX4 pathways.展开更多
目的筛选脑泰方主要有效成分,预测其活性成分干预脑梗死的靶点及通路,对比脑梗死有效靶点及涉及的通路,探究脑泰方干预脑梗死的作用机制。方法采用检索中药系统药理学技术平台(traditional Chinese medicine systems pharmacology datab...目的筛选脑泰方主要有效成分,预测其活性成分干预脑梗死的靶点及通路,对比脑梗死有效靶点及涉及的通路,探究脑泰方干预脑梗死的作用机制。方法采用检索中药系统药理学技术平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)、台湾中医药资料数据库以及文献资料筛选出脑泰方的主要有效成分,利用其活性成分基于Pubchem数据库和Swiss Target Prediction数据库预测脑泰方有效成分的作用靶点,利用TTD数据库检索出脑梗死涉及的靶点,通过生物学信息注释数据库(DAVID)预测分析相应靶点涉及的通路;然后对比不同途径获得的靶点及通路。结果脑泰方中有40个活性成分,预测得到靶点254个,预测到相关信号通路46条;疾病涉及的靶点26个,相关信号通路6条;其中药物活性成分预测的通路与疾病涉及的通路重合的有5条。结论通过数据对比分析可知,脑泰方干预脑梗死可能是通过调节补体和凝血级联通路、钙离子信号通路、肌动蛋白细胞骨架通路等多途径发挥作用,为进一步阐明脑泰方干预脑梗死具体的作用机制提供了重要参考。展开更多
基金The item is supported by Natural Science Foundation of Hunan Province (No. 98JJY 2027)Young-middle Aged Science and Technological Fund of Hunan Province (NO.00JZYZ145)
文摘Objective:To observe the changes of coagulation and fibrinolysis function in human umbilical cord vein endothelial cells (HUVEC) induced by recombinant human tumor necrosis factor-α (rhT-NF-α) and the effect of Naotaifang extract (脑泰方, NTE) on it. Methods: Cultured HUVEC is randomly divided into six groups:Control group, NTE control group (only 2 g/L NTE), rhTNF-α group (100ug/ L rhTNF-α), and low-dosage, middle-dosage and high-dosage NTE group (100ug/L rhTNF-α and 0. 67 g/L, 2 g/L, 6 g/L NTE). The coagulation activity of frozen-dissolved HUVEC, von Willebrand factor (vWF) content in the conditioned medium and tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor (PAD activity were to be detected after 24 hrs. Results: Compared with the control group, PAI activity were enhanced, vWF release markedly increased in conditioned medium of TNF-α group (P<0. 01) and the frozen-dissolved HUVEC markedly shortens the rabbit plasma prothrombin time, and the above changes could be significantly inhibited by the 3 dosages of NTE (P<0. 05, P<0. 01). Conclusion:NTE is effective in inhibiting the coagulation activity of the HUVEC non-stimulated or stimulated by rhTNF-a to enhance the vWF release, and to adjust fibrinolytic function, and mainly to inhibit the PAI activity.
基金supported by the National Natural Science Foundation of China(No.81774033 and No.81773736)the Hunan Provincial Department of Education-funded Scientific Research Project(No.18C0379 and No.19A378)。
文摘Objective:Our previous research showed that Naotaifang(a compound traditional Chinese herbal medicine)extract(NTE)has clinically beneficial effects on neurological improvement of patients with acute cerebral ischemia.In this study,we investigated whether NTE protected acute brain injury in rats and whether its effects on ferroptosis could be linked to the dysfunction of glutathione peroxidase 4(GPX4)and iron metabolism.Methods:We established an acute brain injury model of middle cerebral artery occlusion(MCAO)in rats,in which we could observe the accumulation of iron in neurons,as detected by Perl’s staining.Using assay kits,we measured expression levels of ferroptosis biomarkers,such as iron,glutathione(GSH),reactive oxygen species(ROS)and malonaldehyde(MDA);further the expression levels of transferrin receptor1(TFR1),divalent metal transporter 1(DMT1),solute carrier family 7 member 11(SLC7 A11)and GPX4 were determined using immunohistochemical analysis,real-time quantitative polymerase chain reaction and Western blot assays.Results:We found that treatment with NTE reduced the expression levels of TFR1 and DMT1,reduced ROS,MDA and iron accumulation and reduced neurobehavioral scores,relative to untreated MCAO rats.Treatment with NTE increased the expression levels of SLC7 A11,GPX4 and GSH,and the number of Nissl bodies in the MCAO rats.Conclusion:Taken together,our data suggest that acute cerebral ischemia induces neuronal ferroptosis and the effects of treating MCAO rats with NTE involved inhibition of ferroptosis through the TFR1/DMT1 and SCL7 A11/GPX4 pathways.
文摘目的筛选脑泰方主要有效成分,预测其活性成分干预脑梗死的靶点及通路,对比脑梗死有效靶点及涉及的通路,探究脑泰方干预脑梗死的作用机制。方法采用检索中药系统药理学技术平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)、台湾中医药资料数据库以及文献资料筛选出脑泰方的主要有效成分,利用其活性成分基于Pubchem数据库和Swiss Target Prediction数据库预测脑泰方有效成分的作用靶点,利用TTD数据库检索出脑梗死涉及的靶点,通过生物学信息注释数据库(DAVID)预测分析相应靶点涉及的通路;然后对比不同途径获得的靶点及通路。结果脑泰方中有40个活性成分,预测得到靶点254个,预测到相关信号通路46条;疾病涉及的靶点26个,相关信号通路6条;其中药物活性成分预测的通路与疾病涉及的通路重合的有5条。结论通过数据对比分析可知,脑泰方干预脑梗死可能是通过调节补体和凝血级联通路、钙离子信号通路、肌动蛋白细胞骨架通路等多途径发挥作用,为进一步阐明脑泰方干预脑梗死具体的作用机制提供了重要参考。
