Objective： To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction （当归补血汤, EDBD） was able to protect cardiomyocytes from oxidative injury of rats induced by hydro...Objective： To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction （当归补血汤, EDBD） was able to protect cardiomyocytes from oxidative injury of rats induced by hydrogen peroxide （H2O2） and its potential mechanism. Methods： Myocardial cells from 1- to 2-day-old neonatal rats were cultured in Dulbecco＇s modified Eagle＇s medium low-glucose and Ham＇s F12 medium （1：1）, and the cellular injury was induced by H2O2. The ultra-filtration extract mixture from Angelica sinensis and Hedysarurn po/ybotrys was given in three doses of 3.75, 7.5, and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （Ml-r） assay. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase （LDH） and creatine kinase （CK） releases in the medium, superoxide dismutase （SOD） activities, and intracellular malondialdehyde （MDA） and myeloperoxidase （MPO） contents. The levels of caspase-3 and heat shock protein 70 （hsp70） mRNA expression in cardiomyocytes were measured by reverse transcription polymerase chain reaction. Results： The EDBD could protect the cardiomyocytes from H202 injury in a dose- dependent manner （3.75, 7.50, and 15.00 mg/mL）. The EDBD could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents, increase SOD activity, up-regulate hsp70 expression, and down-regulate caspase-3 expression. Conclusion： The EDBD has protection on cardiomyocytes injured by H202 through improving cell antioxidant ability, up-regulating hsp70 expression, and inhibiting caspase-3 activity.展开更多
Danggui Buxue Tang(DBT) is a famous Chinese medicinal decoction. Mechanism of DBT action is wide ranging and unclear. Exploring new ways of treatment with DBT is useful. Sprague–Dawley(SD) rats were randomly divided ...Danggui Buxue Tang(DBT) is a famous Chinese medicinal decoction. Mechanism of DBT action is wide ranging and unclear. Exploring new ways of treatment with DBT is useful. Sprague–Dawley(SD) rats were randomly divided into 3 groups including control(NC, Saline), the DBT(at a dose of 8.10 g?kg–1), and blood deficiency(BD)(Cyclophosphamide(APH)-and Cyclophosphamide(CTX)-induced anaemia). A metabolomics approach using Liquid Chromatography-Quadrupole-Time-of-Flight/Mass Spectrometry(LC/Q-TOFMS) was developed to perform the plasma metabolic profiling analysis and differential metabolites were screened according to the multivariate statistical analysis comparing the NC and BD groups, and the hub metabolites were outliers with high scores of the centrality indices. Anaemia disease-related protein target and compound of DBT databases were constructed. The TCMSP, Chem Mapper and STITCH databases were used to predict the protein targets of DBT. Using the Cytoscape 3.2.1 to establish a phytochemical component–target protein interaction network and establish a component, protein and hub metabolite protein–protein interaction(PPI) network and merging the three PPI networks basing on BisoGenet. The gene enrichment analysis was used to analyse the relationship between proteins based on the relevant genetic similarity by ClueGO. The results shown DBT effectively treated anaemia in vivo. 11 metabolic pathways are involved in the therapeutic effect of DBT in vivo; S-adenosyl-L-methionine, glycine, L-cysteine, arachidonic acid(AA) and phosphatidylcholine(PC) were screened as hub metabolites in APH-and CTX-induced anaemia. A total of 288 targets were identified as major candidates for anaemia progression. The gene-set enrichment analysis revealed that the targets are involved in iron ion binding, haemopoiesis, reactive oxygen species production, inflammation and apoptosis. The results also showed that these targets were associated with iron ion binding, haemopoiesis, ROS production, apoptosis, inflammation and related signall展开更多
基金Supported by the Key Projects of the National Science and Technology Pillar Program during the Eleventh Five-Year Plan Period(No.2007BAI37B01)Major Science and Technology Program of Gansu Province(No.092NKDA017)
文摘Objective： To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction （当归补血汤, EDBD） was able to protect cardiomyocytes from oxidative injury of rats induced by hydrogen peroxide （H2O2） and its potential mechanism. Methods： Myocardial cells from 1- to 2-day-old neonatal rats were cultured in Dulbecco＇s modified Eagle＇s medium low-glucose and Ham＇s F12 medium （1：1）, and the cellular injury was induced by H2O2. The ultra-filtration extract mixture from Angelica sinensis and Hedysarurn po/ybotrys was given in three doses of 3.75, 7.5, and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （Ml-r） assay. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase （LDH） and creatine kinase （CK） releases in the medium, superoxide dismutase （SOD） activities, and intracellular malondialdehyde （MDA） and myeloperoxidase （MPO） contents. The levels of caspase-3 and heat shock protein 70 （hsp70） mRNA expression in cardiomyocytes were measured by reverse transcription polymerase chain reaction. Results： The EDBD could protect the cardiomyocytes from H202 injury in a dose- dependent manner （3.75, 7.50, and 15.00 mg/mL）. The EDBD could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents, increase SOD activity, up-regulate hsp70 expression, and down-regulate caspase-3 expression. Conclusion： The EDBD has protection on cardiomyocytes injured by H202 through improving cell antioxidant ability, up-regulating hsp70 expression, and inhibiting caspase-3 activity.
基金supported by the National Natural Science Foundation of China(Nos.31560709 and 31472234)the College of Veterinary Medicine of Gansu Agricultural University(No.JYCX-KX005)
文摘Danggui Buxue Tang(DBT) is a famous Chinese medicinal decoction. Mechanism of DBT action is wide ranging and unclear. Exploring new ways of treatment with DBT is useful. Sprague–Dawley(SD) rats were randomly divided into 3 groups including control(NC, Saline), the DBT(at a dose of 8.10 g?kg–1), and blood deficiency(BD)(Cyclophosphamide(APH)-and Cyclophosphamide(CTX)-induced anaemia). A metabolomics approach using Liquid Chromatography-Quadrupole-Time-of-Flight/Mass Spectrometry(LC/Q-TOFMS) was developed to perform the plasma metabolic profiling analysis and differential metabolites were screened according to the multivariate statistical analysis comparing the NC and BD groups, and the hub metabolites were outliers with high scores of the centrality indices. Anaemia disease-related protein target and compound of DBT databases were constructed. The TCMSP, Chem Mapper and STITCH databases were used to predict the protein targets of DBT. Using the Cytoscape 3.2.1 to establish a phytochemical component–target protein interaction network and establish a component, protein and hub metabolite protein–protein interaction(PPI) network and merging the three PPI networks basing on BisoGenet. The gene enrichment analysis was used to analyse the relationship between proteins based on the relevant genetic similarity by ClueGO. The results shown DBT effectively treated anaemia in vivo. 11 metabolic pathways are involved in the therapeutic effect of DBT in vivo; S-adenosyl-L-methionine, glycine, L-cysteine, arachidonic acid(AA) and phosphatidylcholine(PC) were screened as hub metabolites in APH-and CTX-induced anaemia. A total of 288 targets were identified as major candidates for anaemia progression. The gene-set enrichment analysis revealed that the targets are involved in iron ion binding, haemopoiesis, reactive oxygen species production, inflammation and apoptosis. The results also showed that these targets were associated with iron ion binding, haemopoiesis, ROS production, apoptosis, inflammation and related signall
