Objective:To establish a novel cardiocentesis method for withdrawing venous blood from the right atrium,and to improve an acute blood stasis rat model using an ice bath and epinephrine hydrochloride(Epi)while consider...Objective:To establish a novel cardiocentesis method for withdrawing venous blood from the right atrium,and to improve an acute blood stasis rat model using an ice bath and epinephrine hydrochloride(Epi)while considering the 3Rs(reduction,refinement,and replacement)of humane animal experimentation.Methods:An acute blood stasis model was established in male Sprague-Dawley rats by subcutaneous injection(s.c.)Epi(1.2mg/kg)administration at 0h,followed by a 5-min exposure to an ice-bath at 2h and s.c.Epi administration at 4h.Control rats received physiological saline.Rats were fasted overnight and treated with Angelicae Sinensis Lateralis Radix(ASLR)and Pheretima the following day.Venous blood was collected using our novel cardiocentesis method and used to test whole blood viscosity(WBV),prothrombin time(PT),activated partial thromboplastin time(APTT),and fibrinogen(FIB)8ntent.Results:The rats survived the novel cardiocentesis technique;WBV value returned to normal while hematological parameters such as hemoglobin level and red blood cell count were restored to>94%of the corresponding values in normal rats following a 14-day recovery.Epi(1.2 mg/kg,s.c.)combined with a 5-min exposure to the ice bath replicated the acute blood stasis rat model and was associated with the highest WBV value.In rats showing acute blood stasis,ASLR treatment[4g/(kg-d)for 8 days]decreased WBV by 9.98%,11.09%,9.34%,9.00%,7.66%,and 7.03%(P<0.05),while Pheretima treatment[2.6g/(kg?d),for 8 days]decreased WBV by 25.49%,25.94%,16.28%,17.76%,11.07%,and 7.89%(P<0.01)at shear rates of 1,3,10,30,100,and 180 s'1,respectively.Furthermore,Pheretima treatment increased APTT significantly(P<0.01).Conclusions:We presented a stable,reproducible,and improved acute blood stasis rat model,which could be applied to screen drugs for promoting blood circulation and eliminating blood stasis.展开更多
Objective: To compare the pharmacokinetic differences of the main components of Naoxintong capsule(NXTC) in normal and acute blood stasis rats. Materials and Methods: Rats were subcutaneously injected with adrenaline ...Objective: To compare the pharmacokinetic differences of the main components of Naoxintong capsule(NXTC) in normal and acute blood stasis rats. Materials and Methods: Rats were subcutaneously injected with adrenaline hydrochloride twice;during the two subcutaneous injections, the rats were placed in ice water for 4 min to reproduce the model rat of acute blood stasis. The normal and acute blood stasis rats were administrated a 5.04 g/kg dose of NXTC suspension. Then, blood samples were collected from the posterior retinal venous plexus at different time points. Plasma concentrations of four major bio-active components including caffeic acid, ferulic acid, formononetin, and tanshinone IIA in NXTC were measured using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry. Phoenix Win Nonlin v6.2 software was used to calculate the pharmacokinetic parameters. Results: Compared with the normal rats, the acute blood stasis rats showed a significant decrease in C_(max) of ferulic acid and formononetin, AUC_(all) of caffeic acid and ferulic acid, and AUC_(INF_obs) of ferulic acid. Conversely, an increase in the Vz_F_obs and MRT_(last) of ferulic acid and caffeic acid was observed. These findings demonstrate that the absorption of the four NXTC components was weakened in the acute blood stasis rats and that the elimination time was prolonged. Conclusions: The significant difference in some parameters of the four NXTC components between the normal and acute blood stasis rats might be caused by an increase in blood viscosity and the subsequent slowing down of blood flow in the acute blood stasis rats. The pharmacokinetic study conducted in pathological state can provide important information and scientific basis for further rational clinical application of NXTC.展开更多
The traditional Chinese medicine concepts of "Xinxueyuzuzheng (heart blood stasis obstruction pattern)" and "Qiyinliangxuzheng (qi and yin deficiency pattern)" for myocardial ischemia rat models we...The traditional Chinese medicine concepts of "Xinxueyuzuzheng (heart blood stasis obstruction pattern)" and "Qiyinliangxuzheng (qi and yin deficiency pattern)" for myocardial ischemia rat models were constructed in the present study. Endogenous metabolites in rat plasma were analyzed using the GC/TOF-MS-based metabonomic method. Significant metabolic differences were observed between the control and two model groups, and the three groups were distinguished clearly by pattern recognition. Compared with those of the control, the levels of hydroxyproline, threonic acid, glutamine and citric acid were strikingly up or down-regulated in model rats. The metabolites contributing most to the classification between the two "pattern" rats were identified, such as valine, serine, threonine, ornithine, hydroxyproline, lysine, 2-hydroxybutanoic acid, 3-hydroxybutanoic acid, galactofuranose and inositol. These compounds were indicated as the potential biomarkers. The results suggested that the two "patterns" are involved in dysfunction in oxidative stress, energy metabolism and amino acid metabolism. These findings also provided the substantial foundation for exploring the scientific connotation of these two "Zhengxing (pattern types)" of myocardial ischemia, and "Bianzheng (pattern identification)".展开更多
目的:对大鼠急性心肌梗死血瘀证心肌组织代谢组学进行生物信息学分析。方法:在前期实验得出的代谢谱基础上,采用KEGG数据库进行信号通路分析,用HMDB数据库对代谢产物分子注释、相关的酶或转运蛋白及其相关性质进行分析,met PA网络软件...目的:对大鼠急性心肌梗死血瘀证心肌组织代谢组学进行生物信息学分析。方法:在前期实验得出的代谢谱基础上,采用KEGG数据库进行信号通路分析,用HMDB数据库对代谢产物分子注释、相关的酶或转运蛋白及其相关性质进行分析,met PA网络软件对代谢产物路径进行可视化作图。结果:用Met PA分析的生物代谢通路显示,11个代谢产物参与了24条代谢路径。其中柠檬酸循环(Citrate cycle)、丙酮酸代谢(Pyruvate metabolism)、氨酰-tRNA合成(Aminoacyl-tRNA biosynthesis)、半乳糖代谢(Galactose metabolism)、泛醌等萜醌生物合成(Ubiquinone and other terpenoidquinone biosynthesis)、苯丙氨酸,酪氨酸和色氨酸生物合成(Phenylalanine,tyrosine and tryptophan biosynthesis)、D-谷氨酰胺和D-谷氨酸代谢(D-Glutamine and D-glutamate metabolism)、生物素代谢(Biotin metabolism)、酪氨酸代谢(Tyrosine metabolism)、精氨酸和脯氨酸代谢(Arginine and proline metabolism)、氰基氨基酸代谢(Cyanoamino acid metabolism)通路影响值有统计学意义(P<0.05)。结论:大鼠急性心肌梗死血瘀证心肌组织代谢组学显示其病理过程涉及到糖蛋白质最终共同通路的三羧酸循环、氨基酸合成、转运等方面。展开更多
基金Supported by the National Scie nee and Tech no logy Major Project for Major New Drugs Innovation and Development of China(No.2009ZX09502-023-4)National Technology Support Program(No.2006BAI09B06)。
文摘Objective:To establish a novel cardiocentesis method for withdrawing venous blood from the right atrium,and to improve an acute blood stasis rat model using an ice bath and epinephrine hydrochloride(Epi)while considering the 3Rs(reduction,refinement,and replacement)of humane animal experimentation.Methods:An acute blood stasis model was established in male Sprague-Dawley rats by subcutaneous injection(s.c.)Epi(1.2mg/kg)administration at 0h,followed by a 5-min exposure to an ice-bath at 2h and s.c.Epi administration at 4h.Control rats received physiological saline.Rats were fasted overnight and treated with Angelicae Sinensis Lateralis Radix(ASLR)and Pheretima the following day.Venous blood was collected using our novel cardiocentesis method and used to test whole blood viscosity(WBV),prothrombin time(PT),activated partial thromboplastin time(APTT),and fibrinogen(FIB)8ntent.Results:The rats survived the novel cardiocentesis technique;WBV value returned to normal while hematological parameters such as hemoglobin level and red blood cell count were restored to>94%of the corresponding values in normal rats following a 14-day recovery.Epi(1.2 mg/kg,s.c.)combined with a 5-min exposure to the ice bath replicated the acute blood stasis rat model and was associated with the highest WBV value.In rats showing acute blood stasis,ASLR treatment[4g/(kg-d)for 8 days]decreased WBV by 9.98%,11.09%,9.34%,9.00%,7.66%,and 7.03%(P<0.05),while Pheretima treatment[2.6g/(kg?d),for 8 days]decreased WBV by 25.49%,25.94%,16.28%,17.76%,11.07%,and 7.89%(P<0.01)at shear rates of 1,3,10,30,100,and 180 s'1,respectively.Furthermore,Pheretima treatment increased APTT significantly(P<0.01).Conclusions:We presented a stable,reproducible,and improved acute blood stasis rat model,which could be applied to screen drugs for promoting blood circulation and eliminating blood stasis.
基金supported by the National Science and Technology Major Project of China “Key New Drug Creation and Manufacturing Program” 2015ZX09501004-001-007National Natural Science Foundation of China 82004082Top talent training project of TCM in Henan Province。
文摘Objective: To compare the pharmacokinetic differences of the main components of Naoxintong capsule(NXTC) in normal and acute blood stasis rats. Materials and Methods: Rats were subcutaneously injected with adrenaline hydrochloride twice;during the two subcutaneous injections, the rats were placed in ice water for 4 min to reproduce the model rat of acute blood stasis. The normal and acute blood stasis rats were administrated a 5.04 g/kg dose of NXTC suspension. Then, blood samples were collected from the posterior retinal venous plexus at different time points. Plasma concentrations of four major bio-active components including caffeic acid, ferulic acid, formononetin, and tanshinone IIA in NXTC were measured using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry. Phoenix Win Nonlin v6.2 software was used to calculate the pharmacokinetic parameters. Results: Compared with the normal rats, the acute blood stasis rats showed a significant decrease in C_(max) of ferulic acid and formononetin, AUC_(all) of caffeic acid and ferulic acid, and AUC_(INF_obs) of ferulic acid. Conversely, an increase in the Vz_F_obs and MRT_(last) of ferulic acid and caffeic acid was observed. These findings demonstrate that the absorption of the four NXTC components was weakened in the acute blood stasis rats and that the elimination time was prolonged. Conclusions: The significant difference in some parameters of the four NXTC components between the normal and acute blood stasis rats might be caused by an increase in blood viscosity and the subsequent slowing down of blood flow in the acute blood stasis rats. The pharmacokinetic study conducted in pathological state can provide important information and scientific basis for further rational clinical application of NXTC.
基金Supported by the National Natural Science Foundation of China (Grant Nos 30630076 and 30572228)the National 11th 5 Year Technology Support Program (Grant No 2006BAI08B04-05)
文摘The traditional Chinese medicine concepts of "Xinxueyuzuzheng (heart blood stasis obstruction pattern)" and "Qiyinliangxuzheng (qi and yin deficiency pattern)" for myocardial ischemia rat models were constructed in the present study. Endogenous metabolites in rat plasma were analyzed using the GC/TOF-MS-based metabonomic method. Significant metabolic differences were observed between the control and two model groups, and the three groups were distinguished clearly by pattern recognition. Compared with those of the control, the levels of hydroxyproline, threonic acid, glutamine and citric acid were strikingly up or down-regulated in model rats. The metabolites contributing most to the classification between the two "pattern" rats were identified, such as valine, serine, threonine, ornithine, hydroxyproline, lysine, 2-hydroxybutanoic acid, 3-hydroxybutanoic acid, galactofuranose and inositol. These compounds were indicated as the potential biomarkers. The results suggested that the two "patterns" are involved in dysfunction in oxidative stress, energy metabolism and amino acid metabolism. These findings also provided the substantial foundation for exploring the scientific connotation of these two "Zhengxing (pattern types)" of myocardial ischemia, and "Bianzheng (pattern identification)".
文摘目的:对大鼠急性心肌梗死血瘀证心肌组织代谢组学进行生物信息学分析。方法:在前期实验得出的代谢谱基础上,采用KEGG数据库进行信号通路分析,用HMDB数据库对代谢产物分子注释、相关的酶或转运蛋白及其相关性质进行分析,met PA网络软件对代谢产物路径进行可视化作图。结果:用Met PA分析的生物代谢通路显示,11个代谢产物参与了24条代谢路径。其中柠檬酸循环(Citrate cycle)、丙酮酸代谢(Pyruvate metabolism)、氨酰-tRNA合成(Aminoacyl-tRNA biosynthesis)、半乳糖代谢(Galactose metabolism)、泛醌等萜醌生物合成(Ubiquinone and other terpenoidquinone biosynthesis)、苯丙氨酸,酪氨酸和色氨酸生物合成(Phenylalanine,tyrosine and tryptophan biosynthesis)、D-谷氨酰胺和D-谷氨酸代谢(D-Glutamine and D-glutamate metabolism)、生物素代谢(Biotin metabolism)、酪氨酸代谢(Tyrosine metabolism)、精氨酸和脯氨酸代谢(Arginine and proline metabolism)、氰基氨基酸代谢(Cyanoamino acid metabolism)通路影响值有统计学意义(P<0.05)。结论:大鼠急性心肌梗死血瘀证心肌组织代谢组学显示其病理过程涉及到糖蛋白质最终共同通路的三羧酸循环、氨基酸合成、转运等方面。
