RESUMEN
OBJECTIVE: To observe the effect of Kanli Granule (KG) on myocardial mechanics in pressure overload induced diastolic heart failure (DHF) rats. METHODS: Totally 60 male Wistar rats were divided into the sham-operation group, the model group, the KG group, and the Valsartan group according to random digit table, 15 in each group. The pressure overload induced DHF model was established in all groups except the sham-operation group using abdominal aortic constriction surgery. Totally 7 rats died after modeling (with the mortality of 10. 67%) , and the rest 53 finished the following test. Rats in the KG group were administered with KG extract (calculated as 6. 75 g crude drug/kg) by gastrogavage. Rats in the Valsartan group were administered with Valsartan (7.2 µg/g) by gastrogavage. Equal volume of double distilled water was administered to rats in the model group and the sham-operation group by gastrogavage. All rats were intervened for 32 weeks. The response of isolated heart papillary muscle tonus to isoprenaline (ISO) and adenylate cyclase (Forskolin) was respectively observed. The enhancement phenomenon after resting development force (DF) of isolated heart papillary muscle tonus, and changes of DF in different Ca²âº concentrations were observed. RESULTS: (1) In the ISO response test: Compared with the sham-operation group, the amplifications of DF, ±df/dt, -df/dt were obviously elevated in the model group (P < 0.05). Compared with the model group, the amplifications of DF and ±df/dt were obviously lowered in the KG group (P < 0.01), and the amplification of ±df/dt was also reduced in the Valsartan group (P < 0.01). (2) In the Forskolin response test: Compared with the sham-operation group, the amplifications of DF and ±df/dt obviously increased in the model group (P < 0.05). Compared with the model group, the amplifications of DF and ±df/dt were obviously reduced in the KG group (P < 0.01), and the amplification of DF was also reduced in the Valsartan group (P < 0.05). (3) In post-resting DF enhancement test: Compared with the sham-operation group, the amplification of DF showed gradually decreasing tendency along with prolonged resting time in the model group, and they were obviously lowered at all time points (P < 0.05). Compared with the model group, the amplification of DF was gradually increasing along with prolonged resting time in the KG group. The amplification of DF at post-resting 240 s was obviously larger in the KG group than in the model group (P < 0.05). The amplification of post-resting DF still showed gradually decreasing tendency along with prolonged resting time in the Valsartan group, with increased amplifications of DF at post-resting 60 s and 120 s (P < 0. 05) (4) The amplifications of DF in different Ca²âº concentrations: Compared with the sham-operation group, the amplifications of DF were significantly elevated in different Ca²âº concentrations (1.75, 3.5, 7.0 mmol/L ) (P < 0.05, P < 0.01). Compared with the model group, there was no statistical difference in amplification of DF in different Ca²âº concentrations in the KG group (P > 0.05). The amplifications of DF in different Ca²âº concentrations were significantly reduced in the Valsartan group (P < 0.05). CONCLUSIONS: The ISO response and the Forskolin response were enhanced in isolated heart papillary muscle tonus of pressure overload induced DHF rats; enhanced post-resting DF was reduced; DF in different supra-physiologic levels of Ca²âº was still enhanced. KG could significantly improve excessive enhancement of pressure overload induced DHF rats in ISO response and Forskolin response, and improve enhancement of post-resting myocardium.
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Medicamentos Herbarios Chinos/farmacología , Insuficiencia Cardíaca Diastólica/tratamiento farmacológico , Corazón/efectos de los fármacos , Animales , Colforsina/farmacología , Corazón/fisiopatología , Isoproterenol/farmacología , Masculino , Distribución Aleatoria , Ratas , Ratas WistarRESUMEN
Emerging evidence has suggested that inhibitory glycine receptors (GlyRs) are an important molecular target in the treatment of numerous neurological disorders. Rhizoma curcumae is a medicinal plant with positive neurological effects. In this study, we showed that curcumol, a major bioactive component of R. curcumae, reversibly and concentration-dependently inhibited the glycine-activated current (IGly) in cultured rat hippocampal neurons. The inhibitory effect was neither voltage- nor agonist concentration-dependent. Moreover, curcumol selectively inhibited homomeric α2-containing, but not α1- or α3-containing, GlyRs. The addition of ß subunit conferred the curcumol sensitivity of α3-containing, but not α1-containing, GlyRs. Site-directed mutagenesis analysis revealed that a threonine at position 59 of the α2 subunit is critical for the susceptibility of GlyRs to curcumol-mediated inhibition. Furthermore, paralleling a decline of α2 subunit expression during spinal cord development, the degree of IGly inhibition by curcumol decreased with prolonged culture of rat spinal dorsal horn neurons. Taken together, our results suggest that the GlyRs are novel molecular targets of curcumol, which may underlie its pharmaceutical effects in the central nervous system.
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Receptores de Glicina/antagonistas & inhibidores , Sesquiterpenos/farmacología , Secuencia de Aminoácidos , Animales , Células Cultivadas , Sistema Nervioso Central/efectos de los fármacos , Curcuma/química , Interpretación Estadística de Datos , Fenómenos Electrofisiológicos , Femenino , Hipocampo/citología , Hipocampo/efectos de los fármacos , Medicina Tradicional China , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Técnicas de Placa-Clamp , Embarazo , Ratas , Ratas Sprague-Dawley , Receptores de GABA-A/efectos de los fármacos , Receptores de Glicina/química , Receptores de Glicina/efectos de los fármacos , Receptores de Glicina/genética , Proteínas Recombinantes , Treonina/químicaRESUMEN
OBJECTIVE: To study the relationship between the inhibitory effects of Tongxie Yaofang, a compound traditional Chinese herbal medicine, on the contraction of the colonic smooth muscle isolated from rats and calcium mobilization. METHODS: By measuring the tension of the isolated colonic smooth muscle strips, the inhibitory effects of Tongxie Yaofang on the contraction induced by acetylcholine (ACh), KCl and exhausting Ca(2+) of internal calcium store were assessed respectively. RESULTS: Tongxie Yaofang could concentration-dependently inhibit the contraction of isolated rat colonic smooth muscle strips induced by KCl and exhausting the Ca(2+) of internal calcium store. Tongxie Yaofang could also inhibit the tension of the second contractile phase induced by ACh (P<0.01, vs control), but had no influence on the first contractile phase. CONCLUSION: Tongxie Yaofang can inhibit the contraction of isolated rat colonic smooth muscle strips mainly by preventing the influx of extracellular Ca(2+), which may be associated with blocking voltage-dependent channel, store-operated channel and receptor-operated channel, but not by preventing the release of internal Ca(2+) from calcium store.
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Calcio/metabolismo , Colon/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Contracción Muscular/efectos de los fármacos , Animales , Colon/fisiología , Masculino , Músculo Liso , Ratas , Transducción de Señal/efectos de los fármacosRESUMEN
OBJECTIVE: To identify the influence of extracts and active components of Rhizoma Coptidis on gastric smooth muscle contractility of guinea pigs, and to explore the potential pharmacological mechanism of Rhizoma Coptidis in "invigorating the stomach" and "impairing the stomach". METHODS: Observing the effects of the water extract and the alkaloids from Rhizoma Coptidis (at doses ranging from 0.3 to 1,000 microg/L) and other active components such as berberine, palmatine and jatrorrhizine (at doses ranging from 0.3 to 1,000 micromol/L) on the spontaneous and electrical field stimulation (EFS)-induced contractions of antral circular smooth muscle strips from guinea pig stomach via a force transducer in vitro. RESULTS: The water extract or the alkaloids from Rhizoma Coptidis could improve the spontaneous contraction at the low doses, but inhibit the spontaneous contraction at the high doses. Berberine, palmatine and jatrorrhizine also showed the similar effects. Moreover, the water extract and the alkaloids of Rhizoma Coptidis, as well as berberine, palmatine and jatrorrhizine could increase the EFS-induced contraction. Among the three monomers, jatrorrhizine exhibited the most potent effect on EFS-induced contraction. CONCLUSION: The effects of Rhizoma Coptidis in "invigorating the stomach" or "impairing the stomach" may be related to its effect on gastric smooth muscle contractility. Berberine, palmatine and jatrorrhizine are all effective components of Rhizoma Coptidis affecting the contraction of gastric smooth muscle, among which jatrorrhizine is the most potent agent in promoting the contraction while berberine is the most potent one for inhibiting the contraction.
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Medicamentos Herbarios Chinos/farmacología , Músculo Liso/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Berberina/análogos & derivados , Alcaloides de Berberina , Cobayas , Antro PilóricoRESUMEN
OBJECTIVE: To investigate the effects of Astragalus membranaccus (As) on cardiac function and SERCA2a gene expression in left ventricular tissues of rats with chronic heart failure. METHODS: Heart failure was induced by clipping the abdominal aorta 60 male SD rats were divided into four groups: sham-operated (Sham), aortic stenosis (Model), Model + As (20 g/kg) and Model + Captopril (0.05 g/kg). The drugs were administered orally from the 13th week after surgery. Rats were examined after 12 weeks' treatment with drugs. The parameters of hemodynamics including LVSP, LVEDP, and +/- LVdp/dt(max) were measured. SERCA2a mRNA and protein expressions in left ventricular tissues were determined by half-quantitative RT-PCR and Western blot normalized to abundance of GAPDH mRNA and portein, respectively. RESULTS: LVSP and LVEDP were obviously enhanced (P < 0.01 or P < 0.001) in model rats in vivo. Both Captopril and As prevented the increase of LVSP (P < 0.05 or P < 0.01) and LVEDP (P < 0.05 or P < 0.01). RT-PCR and Western blot results demonstrated that SERCA2a gene expression was downregulated (P < 0.05) significantly in model group compared with sham group. As upregulated SERCA2a gene expression (P < 0.05), whereas Captopril had no effect on that. CONCLUSION: As can ameliorate abnormity of cardiac function, especially diastoilc function in rats with pressure overload-induced heart failure, and that may be partly related to its up-regulation of SERCA2a gene expressions in left ventricular tissues.
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Astragalus propinquus , Medicamentos Herbarios Chinos/farmacología , Insuficiencia Cardíaca/tratamiento farmacológico , Corazón/efectos de los fármacos , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/metabolismo , Animales , Astragalus propinquus/química , Captopril/farmacología , Enfermedad Crónica , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/química , Regulación de la Expresión Génica/efectos de los fármacos , Corazón/fisiopatología , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/fisiopatología , Ventrículos Cardíacos/efectos de los fármacos , Ventrículos Cardíacos/metabolismo , Masculino , ARN Mensajero/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/genéticaRESUMEN
OBJECTIVE: To investigate the effect of astragalus (As) on calcium accumulation and SERCA2a gene expression in left ventricular tissues in rats with pressure overload-induced cardiac hypertrophy. METHOD: cardiac hypertrophy was induced by clipping the abdominal aorta in rats. Male SD rats were allocated to six groups: sham-operrated (Sham), aortic stenosis (Model), model +As-L (5 g x kg(-1) x d(-1)), model+As-M (10 g x kg(-1) x d(-1)), model+As-H (20 g x kg(-1) x d(-1)) and model + captopril (0.05 mg x kg(-1) x d(-1), a positive control). The drugs were administered orally from the 13 th week after surgery. Rats were examined after 12 week treatment with drugs. The cardiac hypertrophy was evaluated by left ventricular mass index (LVMI, left ventricular weight/ body weight). The calcium content in left ventricular tissue was measured by atomic absorption spectrometry. SERCA2a mRNA and protein expressions in left ventricular tissues were determined by half-quantitative RT-PCR and Western blot normalized to abundance of GAPDH mRNA and protein, respectively. RESULT: The increase of LVMI was dose-dependently lessened by As (P < 0.01, P < 0.001). The effect of As-H was similar to that of Captopril. As markedly attenuated calcium accumulation in myocardial tissure (P < 0.01). RT-PCR and Western blot results demonstrated that SERCA2a gene expressions were downregulated (P < 0.05) significantly in model group compared with sham group. As-H upregulated SERCA2a gene expressions (P < 0.05), whereas Captopril had no effect on that. CONCLUSION: The inhibition of As on left ventricular hypertrophy induced by pressure overload in rats may partly contribute to its attenuation of calcium accumulation and up-regulation of SERCA2a gene expressions in left ventricular tissues.
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Planta del Astrágalo/química , Calcio/metabolismo , Medicamentos Herbarios Chinos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Hipertrofia Ventricular Izquierda/metabolismo , Miocardio/metabolismo , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/metabolismo , Animales , Western Blotting , Medicamentos Herbarios Chinos/química , Corazón/efectos de los fármacos , Hipertrofia Ventricular Izquierda/fisiopatología , Masculino , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/genéticaRESUMEN
OBJECTIVE: : To profile urinary metabolite variations from 1, 2-dimethylhydrazine (DMH)-induced precancerous colon rats, Jinfu Kang treated rats and healthy controls. METHOD: We used ethyl chloroformate derivatization and gas chromatography-mass spectrometry (GC-MS) based metabonomic method to analyze rat urines. RESULT: The time-dependent variations of metabolite profile showed a progressive deviation of the metabolism in the model group from the initial pattern over time and a systemic recovery of the metabolism in the treatment group, which is consistent with the histological results. The in-depth analysis indicated that the disorder of tricarboxylic acid cycle (TCA), tryptophan metabolism, polyamine metabolism and gut flora structure were associated with DMH intervention. CONCLUSION: Metabolic study revealed that Jinfu Kang can effectively reverse metabolic departures in DMH-induced precancerous colon rat, which is consistent with pathological results.
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Neoplasias del Colon/patología , Pólipos del Colon/tratamiento farmacológico , Pólipos del Colon/orina , Medicamentos Herbarios Chinos/farmacología , Animales , Neoplasias del Colon/inducido químicamente , Pólipos del Colon/inducido químicamente , Dimetilhidrazinas/farmacología , Cromatografía de Gases y Espectrometría de Masas , Masculino , Ratas , Ratas WistarRESUMEN
Fructus Evodiae is a widely used herbal medicine with anti-inflammatory and analgetic activities in China. The present study was designed to investigate the effect of Fructus Evodiae water extract (FE) on ethanol-induced gastric lesions in rats. Three hours before ethanol challenge, animals were intraperitoneally treated with FE (424.8 mg/kg, 141.6 mg/kg, and 47.6 mg/kg). Subsequently, we employed ex-vivo chamber technique to examine the effect of FE on gastric transmucosal potential difference (PD) changes. NO(x) (nitrate and nitrite) in gastric perfusate and gastric lesion index of whole glandular stomach were determined by intubation. The results showed that FE dose-dependently accelerated the recovery of PD reduction by ethanol, and increased NO(x) production in gastric perfusate. FE also inhibited gastric lesion formation in a dose-dependent manner. These results suggested that FE prevented ethanol-induced gastric mucosal lesions by strengthening the mucosal barrier integrity and increasing gastric mucosal nitric oxide (NO) synthesis.
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Medicamentos Herbarios Chinos , Evodia , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patología , Óxido Nítrico/biosíntesis , Animales , Depresores del Sistema Nervioso Central/efectos adversos , Etanol/efectos adversos , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
"Baishouwu" is an appellative name of dried root tubers from three Asclepiadaceae plants: Cynanchum auriculatum Royle ex Wight, Cynanchum bungei Decne and Cynoctonum wilfordii Maxim. In order to establish the pharmacological basis for the ethnomedicinal use of Baishouwu in gastric disorders, this study examined the effects of ethanol extracts and fractions from root tubers of Cynanchum auriculatum, Cynanchum bungei and Cynoctonum wilfordii on ethanol-, indomethacin-induced gastric lesions and histamine-induced gastric acid secretion in rats. Plant materials were collected from various areas of China. Oral administration of ethanol extract and chloroform fraction of Cynoctonum wilfordii collected from Changbai Cordillera at doses of 150 and 68 mg/kg, respectively, significantly inhibited the development of ethanol- and indomethacin-induced gastric lesions and also caused significant decrease of gastric acid secretion after histamine-induced gastric lesion. Oral administrations of ethanol extract and chloroform fraction of Cynanchum auriculatum collected from Binhai at the doses of 300 and 69 mg/kg, respectively, significantly inhibited ethanol- and indomethacin-induced gastric lesions. This study demonstrates the gastroprotective property of Baishouwu for the first time.
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Antiulcerosos/farmacología , Apocynaceae/química , Cynanchum , Medicamentos Herbarios Chinos/farmacología , Mucosa Gástrica/efectos de los fármacos , Úlcera Gástrica/prevención & control , Animales , Antiulcerosos/uso terapéutico , China , Medicamentos Herbarios Chinos/uso terapéutico , Etanol , Etnobotánica , Ácido Gástrico/metabolismo , Determinación de la Acidez Gástrica , Mucosa Gástrica/metabolismo , Histamina , Indometacina , Masculino , Tubérculos de la Planta , Ratas , Ratas Sprague-Dawley , Úlcera Gástrica/inducido químicamenteRESUMEN
OBJECTIVE: To study the proportion and mechanism of relieving asthma of drug partnership comprising herbal Ephedrae & Pheretima. METHOD: To study relaxant effect on 10 micromol x L(-1) carbachol (CCh) and 10 micromol x L(-1) histamine (His) precontracted isolated tracheal rings and lowering effect on short-circuit current (Isc) increase induced by 10 micromol x L(-1) CCh with 3 proportions of 1:1, 1:3, 1:9 extract. RESULT: 1:3 proportions dose-dependently relaxed CCh-precontracted isolated tracheal rings, IC50 of 1:1, 1:3 is 7.5, 15 mg x mL(-1) respectively, 1:9 could not produce 50% inhibition effect on CCh-evoked contraction; 3 proportions also dose-dependently relaxed His-precontracted isolated tracheal rings, IC50 of 1:9, 1:3 and 1:1 is 0.19, 0.61, 1.8 mg x mL(-1) respectively. On the other hand,the orders potency of the decrease effect on CCh-evoked short circuit current increase is 1:3 > 1:1 > 1:9. The difference is not significant (P < 0.05). CONCLUSION: Herbal Ephedrae & Pheretima had tracheal muscle relaxant and epithelium ion secretion inhibition effect, its mechanism of relieving asthma involved anti-CCh and anti-His effect 1:3 was the most appropriate dosage ratio in the anti-asthmatic drug partnership.
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Antiasmáticos/farmacología , Medicamentos Herbarios Chinos/farmacología , Ephedra sinica , Materia Medica/farmacología , Relajación Muscular/efectos de los fármacos , Animales , Antiasmáticos/administración & dosificación , Asma/fisiopatología , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/aislamiento & purificación , Ephedra sinica/química , Cobayas , Antagonistas de los Receptores Histamínicos/farmacología , Técnicas In Vitro , Masculino , Materia Medica/administración & dosificación , Materia Medica/aislamiento & purificación , Músculo Liso/efectos de los fármacos , Oligoquetos/química , Plantas Medicinales/química , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To investigate the protective effects of Evodia rutaecarpa water extract on ethanol-induced acute gastric lesions in rats. METHOD: In this study, animals were intraperitoneally pretreated with vehicle (normal saline) or E. rutaecarpa at 424.8, 141.6, 47.2 mg x kg(-1). Three hours later, gastric lesions were induced by topical application of 50% ethanol for 10 min. The rat gastric transmucosal potential difference (PD) was recorded continuously by ex-vivo chamber technique. NO(x) (nitrate and nitrite) level in gastric perfusate and the length index of gastric lesions were determined in a gastric intubatton model. RESULT: Compared with control, Evodia rutaecarpa water extract accelerated PD recovery and reduced gastric morphologic lesions in a dose-dependent manner (P < 0.05) , and also dose-dependently increased NO(x) level in gastric perfusate. Especially, at 424.8 mg x kg(-1), E. rutaecarpa promoted synthesizing of nitric oxide significantly (P < 0. 01). CONCLUSION: E. rutaecarpa water extract showed effectively protective actions on ethanol-induced acute gastric lesions in rats, and the gastroprotective mechanisms maybe due to the strengthening action on gastric mucosal lining and the promotion of nitric oxide synthesis in local gastric mucosa.
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Medicamentos Herbarios Chinos/farmacología , Evodia , Sustancias Protectoras/farmacología , Úlcera Gástrica/patología , Animales , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/aislamiento & purificación , Etanol , Evodia/química , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patología , Masculino , Óxido Nítrico/biosíntesis , Plantas Medicinales/química , Sustancias Protectoras/administración & dosificación , Sustancias Protectoras/aislamiento & purificación , Ratas , Ratas Sprague-Dawley , Úlcera Gástrica/inducido químicamente , Úlcera Gástrica/metabolismoRESUMEN
The purpose of the present study was to examine the effects of calycosin, an isoflavonoid isolated from Astragali Radix, on the impairment of barrier function induced by hypoxia in cultured human umbilical vein endothelial cells. Hypoxia induced an increase in endothelial cell monolayer permeability, indicating endothelial cell barrier impairment. Endothelial barrier dysfunction induced by hypoxia was accompanied by decreases in cytosolic ATP concentration and cAMP level, the development of actin stress fibers and intercellular gap formation, suggesting that the decreases in cytosolic ATP and cAMP levels and rearrangements of F-actin could be associated with an increase in permeability of endothelial monolayers. Application of calycosin inhibited the hypoxia-induced increase in endothelial permeability in a dose-dependent fashion, which is compatible with inhibition of lactate dehydrogenase release, decrease of the fall in ATP and cAMP contents, and improvement of F-actin rearrangements. These findings indicate that calycosin protected endothelial cells from hypoxia-induced barrier impairment by increasing intracellular energetic sources and promoting regeneration of the cAMP level, as well as improving cytoskeleton remodeling.
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Hipoxia de la Célula , Células Endoteliales/efectos de los fármacos , Fluoresceína-5-Isotiocianato/análogos & derivados , Isoflavonas/farmacología , Citoesqueleto de Actina/metabolismo , Adenosina Trifosfato/metabolismo , Permeabilidad de la Membrana Celular/efectos de los fármacos , Células Cultivadas , AMP Cíclico/metabolismo , Citosol/metabolismo , Dextranos/química , Dextranos/metabolismo , Medicamentos Herbarios Chinos/farmacología , Células Endoteliales/metabolismo , Células Endoteliales/fisiología , Fluoresceína-5-Isotiocianato/química , Fluoresceína-5-Isotiocianato/metabolismo , Humanos , L-Lactato Deshidrogenasa/metabolismo , Peso Molecular , Venas Umbilicales/citología , Venas Umbilicales/efectos de los fármacos , Venas Umbilicales/fisiopatologíaRESUMEN
OBJECTIVE: TO establish a RP-HPLC method for the determination of calycosin-7-O-beta-D-glucopyranoside in Radix Astragali, and to analyse the calycosin-7-O-beta-D-glucopyranoside content of ten samples of Radix Astragali, collected from different regions. METHOD: A Polaris C18(250 mm x 4.6 mm, 5 microns) column was used and a mixture of methanol-water (30:70) was used as the mobile phase at a flow rate of 1.0 mL.min-1. The column temperature was 25 degrees C and the UV detection wavelength was 254 nm. RESULT: The calibration curve was in good linearity over the range of 0.0106-2.12 micrograms with the regression equation Y = 3035. 97 X - 14.85(r = 0.9999). The average recovery was 95.8% (n = 5, RSD = 1.3%). CONCLUSION: The method is simple, quick, sensitive and reproducible. In all of the samples, the calycosin-7-O-beta-D-glucopyranoside contents differ markedly.