Two cardenolide glycosides, corotoxigenin 3-O-[β-D-glucopyranosyl-(1→4)-6-deoxy-β-D-glucopyranoside] (1) and coroglaucigenin 3-O-[β-D-glucopyranosyl-(1→4)-6-deoxy-β-D-glucopyranoside] (2), were isolated from the seed fairs of Asclepias curassavica. The structures of 1-2 were determined based on the combination of the analysis of their MS, NMR spectroscopic data and acid hydrolysis. The inhibitory effects of compounds 1 and 2 on human colorectal carcinoma cells (HCT116), non-small cell lung carcinoma cells (A549) and hepatic cancer cells (SMMC-7721) were evaluated. The results showed that both compounds 1 and 2 significantly inhibited the viability, proliferation, and migration of A549, HCT116 and SMMC-7721 cells, suggesting that compounds 1 and 2 can be applied in the treatment of lung, colon and liver cancers in clinical practice. This study may not only provide a scientific basis for clarifying the active ingredients in A. curassavica, but also help to understand its antitumor activity, which can promote the application of A. curassavica in clinical treatment of various cancers.
Antineoplastic Agents/pharmacology* ; Asclepias/chemistry* ; Cardenolides/pharmacology* ; Glycosides/pharmacology* ; Humans ; Seeds

AIMTo study the effect of antidigoxin antiserum on oxygen stress induced by myocardial ischemia/reperfusion (MI/R) injury in rats.

METHODSSprauge Dawley rats were submitted to ligate left anterior descending coronary artery 30 min followed by 45 min reperfusion. Experiment animals were randomly divided into seven groups including sham group, MI/R group, normal salina group, verapamil group and three antidigoxin antiserum groups from low to high dose. The left ventricular myocardial tissue sample of ischemia were processed and measured the level of endoxin and malondialdehyde (MDA), the activities of Na+, K(+) -ATPase and superoxin dismutase (SOD). The myocardia morphology was observed.

RESULTSThe levels of endoxin and MDA increased and the activities of Na+, K(+) -ATPase and MDA were inhibited significantly in MI/R and saline groups. Including verapamil group in comparison to MI/R and saline groups, MDA level decreased and SOD activities partly reserved, meanwhile, only in three antidigoxin antiserum groups, the myocardial endoxin level was remarkably decreased, Na+, K(+) -ATPase activities were drastically increased. The myocardial histological morphology was significantly improved.

CONCLUSIONAntidigoxin antiserum, an endoxin mutual clone antibody, had the effect of attenuating the damage of oxygen free radicals induced by MI/R via to antagonizing the inhibition effect of endoxin on myocardial membrane Na+, K(+) -ATPase activities.

Animals ; Cardenolides ; antagonists & inhibitors ; Digoxin ; pharmacology ; Immune Sera ; pharmacology ; Malondialdehyde ; analysis ; Myocardial Reperfusion ; Myocardial Reperfusion Injury ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Saponins ; antagonists & inhibitors ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism

To study the anticancer activity of griffithin from Streptocaulon griffithii Hook. f. and its effect on apoptosis of cancer cells in vitro, the inhibitory effect of griffithin on cell proliferation was studied by MTT assay, the cell apoptosis was observed by AO/EB double decoration assay and flow cytometry. Griffithin exhibited high anticancer activity on four human cancer cell lines, with IC50 ranged from 0.17 - 0.43 microg x mL(-1). Griffithin also induced apoptosis of PC-3 cells. Griffithin had anticancer activity and induced apoptosis of cancer cells.
Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Apocynaceae ; chemistry ; Apoptosis ; drug effects ; Cardenolides ; administration & dosage ; chemistry ; isolation & purification ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Flow Cytometry ; HL-60 Cells ; Humans ; Inhibitory Concentration 50 ; Microscopy, Fluorescence ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

BACKGROUNDEndogenous hydrogen sulfide is a new neuromodulator which takes part in the regulation of central nervous system physiology and diseases. Whether endogenous hydrogen sulfide in the central nervous system regulates cardiovascular activity is not known. In the present study, we observed the hemodynamic changes of hydrogen sulfide or its precursor by intracerebroventricular injection, and investigate the possible roles of endogenous digitalis like factors and sympathetic activity in the regulation.

METHODSNinety-four Sprague-Dawley rats underwent a right cerebroventricular puncture, then the hydrogen sulfide saturation buffer or its precursor injected by intrcerebroventricular catheter. A heperin-filled catheter was inserted into the right femoral artery or into the left ventricle, and changes of blood pressure or cardiac function recorded by a Powerlab/4S instrument. Phentolamine or metoprolol were pre-injected to observe the possible role in autonomic nerve activity. After rats were sacrificed, plasma was collected and endogenous digitalis-like factors were measured with a commercial radioimmunoassay kit. The aortic, cardiac sarcolemmal vesicles were isolated and the activity of Na(+)-K(+)-ATPase was measured as ouabain-sensitive ATP hydrolysis under maximal velocity conditions by measuring the release of inorganic phosphate from ATP. Unpaired Student's t test for two groups or analysis of variances (ANOVA) for multiple groups were used to compare the differences of the changes.

RESULTSIntracerebroventricular injection of hydrogen sulfide induced a transient hypotension, then dramatic hypertenive effects in a dose-dependent manner. Bolus injection of L-cysteine or beta- mercaptopyruvate also increased mean arterial pressure (P < 0.01), whereas hydroxylamine-a cystathionine beta synthase inhibitor decreased the arterial pressure (P < 0.01). Hydrogen sulfide and L-cysteine increased mean arterial pressure, left ventricular develop pressure and left-ventricle maximal rate of systolic and diastolic pressure; these functions were decreased by hydroxylamine (P < 0.01). Glibenclamide (a K(ATP) channel blocker) blocked the transient hypotensive effect, phentolamine (an alpha-adrenergic receptor blocker) blocked the hypertensive effect, and metoprolol (a selective beta 1 receptor blocker) blocked the positive inoptropic effect of central nervous system hydrogen sulfide. The endogenous digitalis-like factors in plasma were elevated (P < 0.01) after treatment with L-cysteine, association with decreasing Na(+)-K(+)-ATPase activity in cardiac or aortic sarcolemmal vesicles (P < 0.01). Hydroxylamine injection reduced the endogenous digitalis-like factors level in plasma association with increasing Na(+)-K(+)-ATPase activity in cardiac and aortic sarcolemmal vesicles.

CONCLUSIONCentral nervous system endogenous hydrogen sulfide upregulated mean arterial pressure and cardiac systolic function by activation of sympathetic nerves or release of endogenous digitalis-like factors.

Animals ; Blotting, Western ; Cardenolides ; metabolism ; Central Nervous System ; drug effects ; metabolism ; Cystathionine beta-Synthase ; metabolism ; Cysteine ; analogs & derivatives ; pharmacology ; Hemodynamics ; drug effects ; Hydrogen Sulfide ; metabolism ; pharmacology ; Male ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley ; Saponins ; metabolism ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Sulfurtransferases ; metabolism

Although Nerium indicum poisoning is a globally rare occurrence, Nerium oleander poisoning is known to occur frequently in the Mediterranean regions. To our knowledge, this is the first reported case of accidental Nerium indicum poisoning in Korea. Its poisoning symptoms and signs are similar to that of digitalis poisoning, because of the presence of cardiac glycosides in Nerium indicum. A 16-year-old boy was admitted to the emergency department four hours prior to the accidental ingestion of Nerium indicum petals. The patient complained of nausea, vomiting, and dizziness. His initial vital signs were stable; laboratory blood test results were within normal levels, except for the blood digoxin level (1.5 ng/dL). An electrocardiogram (ECG) analysis showed normal sinus rhythm, progressive PR prolongation and second-degree Morbiz type I AV block. Conservative treatments including activated charcoal administration were conducted, because toxic symptoms and signs were not severe. The patient was admitted to the intensive care unit for close observation. His ECG was converted to normal rhythm after 1 day and the toxic symptoms and signs were completely resolved after 4 days.
Adolescent ; Atrioventricular Block ; Cardiac Glycosides ; Charcoal ; Digitalis ; Digoxin ; Dizziness ; Eating ; Electrocardiography ; Emergencies ; Hematologic Tests ; Humans ; Intensive Care Units ; Korea ; Mediterranean Region ; Nausea ; Nerium ; Vital Signs ; Vomiting

BACKGROUND: Purification of opioid receptor is mandatory to improve opiate analgesic medication. Recently, it was reported that sodium ion increased the number of opioid binding sites for opioid antagonist. The importance of sodium ions lead us to design appropriate affinity chromatography and binding assay for the successful purification of mu-opioid receptor to homogeneity. METHODS: Opioid receptor was solubilized from rat brain membranes with a mixture of the detergents, CHAPS and digitonin, in the presence of protease inhibitors and 1M NaCl. The solubilized material was passed through an opioid antagonist(10cd) affinity column and a wheat germ agglutinin(WGA) column, set up in series, to obtain a partially purified receptor preparation. The partially purified receptor was further purified by repeating the affinity and lectin chromatography with smaller size column. RESULTS: Binding of opioid antagonist [H]diprenorphine to the partially purified or purified receptors was dependent upon the presence of sodium ions. The purified receptor showed diffuse band with a medium molecular mass of 62KD upon electrophoresis. The average specific binding activity of the purified receptor was 18.8+/-2.3 pmol/mcg protein. CONCLUSIONS: Opioid agonists and antagonists either do not bind or bind with low affinity to G protein-dissociated free opioid receptors in the absence of sodium ions. However, the free opioid receptors have a high affinity for antagonists but not agonists in the presence of sodium ions.
Animals ; Binding Sites ; Brain ; Chromatography ; Chromatography, Affinity ; Detergents ; Digitonin ; Electrophoresis ; Ions ; Membranes ; Protease Inhibitors ; Rats ; Receptors, Opioid* ; Sodium* ; Triticum

OBJECTIVE: To compare the clinical efficacy between Wei's triple nine needling combined with esculin and digitalis glycosides eye drops and esculin and digitalis glycosides eye drops alone for presbyopia complicated with visual fatigue of liver depression and spleen deficiency. METHODS: Forty-six cases (92 eyes) with presbyopia complicated with visual fatigue of liver depression and spleen deficiency were randomly divided into an observation group (23 cases) and a control group (23 cases, 2 cases dropped off). The cases in the observation group were treated with Wei's triple nine needling and esculin and digitalis glycosides eye drops. The acupoints included Shangming (Extra), Chengqi (ST 1), Cuanzhu (BL 2) to Jingming (BL 1), Sizhukong (TE 23) to Taiyang (EX-HN 5), etc; the needling was given once every other day, three times a week, and the eye drops were given one drop each time, three times a day. The cases in the control group were only treated with the eye drops. Both groups were treated for 7 days as one course of treatment, and 2 courses of treatment were given. The visual fatigue core symptoms score, adjustment amplitude, adjustment lag and best average corrected visual acuity were observed in the two groups before treatment, 1 week and 2 weeks into treatment, respectively. RESULTS: Compared before treatment, the visual fatigue core symptoms scores in the two groups were decreased after 1-week and 2-week treatment (P<0.05); in the observation group, the adjustment amplitude was increased after 2-week treatment (P<0.05), while in the control group, the adjustment amplitude was increased after 1-week and 2-week treatment (P<0.05); in the observation group, the adjustment lag was decreased after 1-week and 2-week treatment (P<0.05). After 2-week treatment, the visual fatigue core symptoms score in the observation group was lower than that in the control group, and the adjustment amplitude was higher than that in the control group (P<0.05). There were no significant differences in adjustment lag and best average corrected visual acuity between the two groups after 1-week and 2-week treatment (P>0.05). CONCLUSION Wei's triple nine needling combined with esculin and digitalis glycosides eye drops could improve the visual fatigue and eye regulation ability in patients with presbyopia complicated with visual fatigue of liver depression and spleen deficiency, and the effect is better than esculin and digitalis glycosides eye drops alone.
Acupuncture Points ; Acupuncture Therapy ; Asthenopia ; Depression ; Digitalis Glycosides ; Esculin ; Humans ; Liver ; Ophthalmic Solutions ; Presbyopia ; Spleen ; Treatment Outcome

Influences of intracerebroventricular(icv) ouabain on cardiovascular system and on pressor response to raised intracranial pressure(ICP) were investigated in urethane-anesthetized rabbits. Intravenous(iv) 40 microgram/kg ouabain did not affect blood pressure of rabbit. Icv ouabain(5, 10, 20 microgram/kg) and digoxin(6.25, 12.5 microgram/kg) produced dose-dependent pressor responses and marked decrease of heart rate. After bilateral vagotomy, the bradycardiac effect of ouabain disappeared, while the pressor effect was significantly potentiated. The ouabain-induced pressor effect was not affected by iv and icy phentolamine or propranolol, and significantly inhibited by icy infusion of lidocaine(0.1 mg/kg/min) and diazepam(0.025 mg/kg/min). The pressor response to raised ICP was not affected by iv 40 microgram/kg ouabain but disappeared by icy 10 microgram/kg ouabain and 6.25 microgram/kg digoxin. These data suggests that ouabain causes pressor effect through direct interaction with a site(s) in central nervous system and bradycardiac effect by direct activation of vagal center, and ouabain blocks central sympathetic stimulation by increase of ICP in rabbits.
Blood Pressure ; Cardiovascular System* ; Central Nervous System ; Digitalis* ; Digoxin ; Heart Rate ; Intracranial Pressure ; Ouabain ; Phentolamine ; Propranolol ; Rabbits* ; Vagotomy

Mammalian acetyl-CoA carboxylase (ACC) is present in two isoforms, alpha and beta, both of which catalyze formation of malonyl-CoA by fixing CO2 into acetyl-CoA. ACC-alpha is highly expressed in lipogenic tissues whereas ACC-beta is a predominant form in heart and skeletal muscle tissues. Even though the tissue-specific expression pattern of two ACC isoforms suggests that each form may have a distinct function, existence of two isoforms catalyzing the identical reaction in a same cell has been a puzzling question. As a first step to answer this question and to identify the possible role of ACC isoforms in myogenic differentiation, we have investigated in the present study whether the expression and the subcellular distribution of ACC isoforms in H9c2 cardiac myocyte change so that malonyl-CoA produced by each form may modulate fatty acid oxidation. We have observed that the expression levels of both ACC forms were correlated to the extent of myogenic differentiation and that they were present not only in cytoplasm but also in other subcellular compartment. Among the various tested compounds, short-term treatment of H9c2 myotubes with insulin or okadaic acid rapidly increased the cytosolic content of both ACC isoforms up to 2 folds without affecting the total cellular ACC content. Taken together, these observations suggest that both ACC isoforms may play a pivotal role in muscle differentiation and that they may translocate between cytoplasm and other subcellular compartment to achieve its specific goal under the various physiological conditions.
Acetyl-CoA Carboxylase/metabolism* ; Acetyl-CoA Carboxylase/drug effects ; Animal ; Cell Differentiation/drug effects ; Cell Line ; Cell Membrane Permeability ; Chromones/pharmacology ; Cytosol/enzymology* ; Cytosol/drug effects ; Digitonin/pharmacology ; Immunoblotting ; Insulin/pharmacology* ; Isoenzymes ; Morpholines/pharmacology ; Myocardium/cytology ; Okadaic Acid/pharmacology* ; Phosphorylation ; Rats

BACKGROUNDOuabain and digoxin are important cardiac glycoside and related to many cardiovascular diseases. The purpose of this study was to investigate the changes of sodium pump α-subunit expression in rats and compare the effects of ouabain (OUA) and digoxin (DIG) on the development of hypertension.

METHODSIn situ hybridization was performed. Specific sequence oligonucleotide probe tailing with a Dig-dUTP hybrid to target nucleic acids of the sodium pump α-subunit. According to counting positive particles sodium pump subunit expression was analyzed with statistical methods.

RESULTSOn day 16 of drug administration, the blood pressure of rats increased significantly in the OUA group. In the DIG group, the blood pressure revealed no significant difference when compared to the control group. In addition, the effects of OUA and DIG on sodium pump α-subunit RNA expression in tissues differed.

CONCLUSIONSOUA and DIG can not only change the configuration of the sodium pump to depress their activity, but also influence their gene expression which is important in the mechanism of hypertension. This may be a key point in the pathogenesis of hypertension in the manner in which OUA differs from DIG and changes the sodium pump gene expression in the arteries and kidneys of rats.

Animals ; Blood Pressure ; drug effects ; Digoxin ; therapeutic use ; Hypertension ; drug therapy ; In Situ Hybridization ; Male ; Ouabain ; therapeutic use ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; metabolism