[ALKALOIDS FROM NUX VOMICA LEAVES: STRYCHNOS NUX VOMICA L (LOGANIACEAE)].
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Alkaloids↗
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Processing of nux vomica. VII. Antinociceptive effects of crude alkaloids from the processed and unprocessed seeds of Strychnos nux-vomica in mice.
We examined the antinociceptive effects of the crude alkaloid fractions (CAF) of nux vomica (the dried seeds of Strychnos nux-vomica L.) and the influences of various processing methods upon their antinociception in three analgesic tests in mice. In the tail-pressure test, the CAF (0.01--1 micrograms/kg, i.p.) of nux vomica that was unprocessed or treated with sand-, licorice-, oil- or vinegar and sand-processing showed clear antinociception. The CAF (1 microgram/kg, i.p.) of vinegar-processed nux vomica showed antinociception, without effects at lower doses of 0.01 and 0.1 microgram/kg and those treated with urine- or urine and sand-processing were without effects at doses of 0.01--1 microgram/kg. Morphine (2 mg/kg, s.c.) showed short-lasting antinociception, without effects at a dose of 1 microgram/kg. In the hot-plate test, the CAF (100 microgram/kg, i.p.) of nux vomica having undergone sand-processing produced a significant antinociception, without effects at lower doses of 0.01 and 1 microgram/kg. The CAF (0.01--100 microgram/kg, i.p.) of nux vomica that was unprocessed or treated with oil- or vinegar and sand-processing and morphine (1 and 100 micrograms/kg, s.c.) were without effects. In the acetic acid-induced writhing test, the CAF (1 microgram/kg, i.p.) of nux vomica that was treated with sand-processing significantly inhibited the writhing behavior, while those of nux vomica that was unprocessed or treated with oil- or vinegar and sand-processing and morphine were without effects at a dose of 1 microgram/kg. The present results demonstrate the antinociceptive effects of the CAF of nux vomica and suggest that sand-processing is good for the analgesic potency of nux vomica. It is also suggested that the CAF of nux vomica has distinct antinociceptive potency, even after treatment with licorice-, oil-, vinegar and sand-processing.
Acetic Acid↗
Processing of nux vomica. II. Changes in alkaloid composition of the seeds of Strychnos nux-vomica on traditional drug-processing.
In the course of our study on the drug-processing of the seeds of Strychnos nux-vomica L. (Loganiaceae), the alkaloid composition of the heat-treated seeds of S. nux-vomica was compared to that of the untreated seeds. On heat treatment, the contents of the major alkaloids such as strychnine and brucine declined significantly with increases in the amounts of isostrychnine, isobrucine, strychnine N-oxide and brucine N-oxide. The cleavage of an ether linkage and the occurrence of N-oxidation were demonstrated by heat treatment of authentic strychnine and brucine.
Alkaloids↗
Altered solution structure of alcoholic medium of potentized Nux vomica underlies its antialcoholic effect.
Nux vomica 30c, 200c and 1000c were administered orally to three batches of albino mice for three days. Six hours after the last dose on the third day the mice were injected i.p. with ethanol 4g/kg body wt. They lost their righting reflex and lay motionless apparently sleeping due to alcohol. Mice treated with three potencies of Nux vomica regained their righting reflex more quickly than the corresponding untreated controls. Each of the three batches of mice was tested twice for ethanol sedation, once with a potency of Nux vomica and another time with a placebo control. The time interval between drug treatment and control was 10 days. NMR spectra of Nux 30, Nux 200, Nux 1000, alcohol 30, alcohol 30 (unagitated) and 90% alcohol showed significant difference from each other with respect to the spin-lattice relaxation time (T1) of the deuterium nuclei. This gives a measurable physical basis of the effective high potencies of Nux vomica.
Administration, Oral↗
Nux vomica 30 prepared with and without succession shows antialcoholic effect on toads and distinctive molecular association.
Adult toads, Bufo melanostictus, were administered Nux vomica (Nux v) 30 prepared with and without succussion on the tongue. The drug was mixed with sterile distilled water at the rate 0.05ml/ml water and given orally 0.05ml/individual. The control consisted of blank ethanol solution. Seeds of Strychnos nuxvomica were ground and extracted with 90% ethanol in the laboratory. Nux v 30 was prepared by successive dilution and succussion in 30 steps, Nux v 30 u was prepared by successive dilution only. Four hours after treatment, toads were given 25% ethanol i.p. at 8g/kg body weight. The duration of ethanol induced sleep time was recorded for each toad. Both Nux v 30 and Nux v 30 u significantly reduced ethanol induced sleep time in toads as compared to their respective controls. Electronic, infra red and nuclear magnetic resonance spectra of Nux v 30, Nux v 30 u and their diluent medium (90% ethanol) show marked differences from each other. These dilutions and ethanol 30 and ethanol 30 u show marked differences from each other with respect to spin-lattice relaxation time (T1) and chemical shift. The difference has been attributed to the variation in intra and inter-molecular association of ethanol and water.
Administration, Oral↗
THE MORPHOLOGY AND HISTOLOGY OF THE SEEDS OF STRYCHNOS NUX-VOMICA, LINN., AND ITS ADULTERANTS, STRYCHNOS NUX-BLANDA, HILL, AND STRYCHNOS POTATORUM, LINN.
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Histology↗
Iridoid glucosides from Strychnos nux-vomica.
From seeds of Strychnos nux-vomica three iridoids, 6'-O-acetylloganic acid, 4'-O-acetylloganic acid and 3'-O-acetylloganic acid were isolated together with two known iridoid glucosides, loganic acid and 7-O-acetylloganic acid. The structures of the compounds were established by ESI-MS and by 1D and 2D NMR spectroscopic methods.
Iridoids↗
Strychnos nux-vomica extract and its ultra-high dilution reduce voluntary ethanol intake in rats.
OBJECTIVES: To see whether Strychnos nux-vomica extract (mother tincture [MT]), its potency Nux 30c, and its principal alkaloid, strychnine, could reduce voluntary ethanol intake in rats. To analyze the solution structure of Nux MT, Nux 30c, 90% ethanol, and ethanol 30c by means of electronic (ES) and nuclear nuclear magnetic resonance (NMR) spectra. DESIGN: Potentially alcoholic rats were first given 20% ethanol and then kept on a two-choice bottle, one with 20% ethanol and another with tap water. These rats were given the following oral treatments for 15 days: group 1, control; group 2, strychnine at 0.36 mg/kg per day; group 3, ethanolic extract of S. nux-vomica seeds (Nux MT) at 3.6 mg/kg per day; and group 4, Nux 30c at 0.05 mL/d per rat. Nux 30c was prepared by successive dilution of Nux MT and 90% ethanol (1:100) and sonication at 20 kHz for 30 seconds in 30 steps. RESULTS: Both Nux MT and Nux 30c significantly reduced ethanol intake and increased water intake in rats. ES of two dilutions of Nux MT and Nux 30c showed intersections at more than one point suggesting existence of molecular complexes. ES of Nux MT in CCl4 showed a red shift when 90% ethanol was added indicating molecular complexation and charge transfer interaction between ethanol and Nux compounds. NMR spectra of Nux MT, 90% ethanol, ethanol 30c, and Nux 30c indicated a change in solution structure of the medium (90% ethanol) of Nux 30c. CONCLUSION: Nux MT and Nux 30c could reduce ethanol intake in rats. The altered solution structure of Nux 30c is thought to mimic Nux MT and produce ethanol aversion in rats.
Alcohol Deterrents↗
Reiter's disease treated with Nux vomica.
We report the case of a 35 year-old man suffering from Reiter's disease. He did not respond to Lycopodium or Nux vomica in medium dilutions, but did respond to Nux vomica in very high potency.
Adult↗
Analysis of strychnine from detoxified Strychnos nux-vomica [corrected] seeds using liquid chromatography-electrospray mass spectrometry.
The content of strychnine from Strychnos nux-vomica [corrected] seeds was analyzed and compared to processed seeds by the HPLC-ESI/MS method. Using this technique, levels as low as 1 ng of strychnine were detected. In contrast to conventional UV detectors, this method also made it possible to discriminate brucine. This study resulted in finding the content of strychnine in detoxified seeds to be one tenth of unprocessed Strychnos nux-vomica [corrected] seeds.
Plant Extracts↗
The apoptotic effect of brucine from the seed of Strychnos nux-vomica on human hepatoma cells is mediated via Bcl-2 and Ca2+ involved mitochondrial pathway.
In an attempt to dissect the mechanism of Strychnos nux-vomica, a commonly used Chinese folk medicine in the therapy of liver cancer, the cytotoxic effects of four alkaloids in Strychnos nux-vomica, brucine, brucine N-oxide, strychnine, and isostrychnine, on human hepatoma cells (HepG2) were screened by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrasolium bromide (MTT) assay. Brucine, among the four alkaloids, exhibited the strongest toxic effect, the mechanism of which was found to cause HepG2 cell apoptosis, since brucine caused HepG2 cell shrinkage, the formation of apoptotic bodies, DNA fragmentation, cell cycle arrest, as well as phosphatidylserine externalization, all of which are typical characteristics of apoptotic programmed cell death. Brucine-induced HepG2 cell apoptosis was caspase dependent, with caspase-3 activated by caspase-9. Brucine also caused the proteolytic processing of caspase-9. In addition, brucine caused depolarization of the mitochondrial membrane of HepG2 cells, the inhibition of which by cyclosporine A completely abrogated the activation of casapses and release of cytochrome c in brucine-treated HepG2 cells. These findings suggested a pivotal role of mitochondrial membrane depolarization in HepG2 cell apoptosis elicited by brucine. Furthermore, brucine induced a rapid and sustained elevation of intracellular [Ca2+], which compromised the mitochondrial membrane potential and triggered the process of HepG2 cell apoptosis. Finally, Bcl-2 was found to predominately control the whole event of cell apoptosis induced by brucine. The elevation of [Ca2+]i caused by brucine was also suppressed by overexpression of Bcl-2 protein in HepG2 cells. From the facts given above, Ca2+ and Bcl-2 mediated mitochondrial pathway were found to be involved in brucine-induced HepG2 cell apoptosis.
Apoptosis↗
Separation and determination of strychnine and brucine in Strychnos nux-vomica L. and its preparation by nonaqueous capillary electrophoresis.
An easy, rapid method for simultaneous determination of strychnine and brucine in Strychnos nux-vomica L. and its preparation was developed by nonaqueous capillary electrophoresis (NACE) without pretreatment for the first time. Optimum separation was achieved with a fused-silica capillary column (50 cmx75 microm i.d.) and a running buffer containing 30 mM ammonium acetate, 1.0% acetic acid and 15% acetonitrile (ACN) in methanol medium. The applied voltage was 30.0 kV. The analytes were detected by UV at 214 nm. The effects of concentration of ammonium acetate, acetic acid and organic modifier on electrophoretic behavior of the analytes were studied. The established method with sophoridine as internal standard was linear in the range of 5-1000 mg/mL for both strychnine and brucine. The extracts of Strychnos nux-vomica and its preparation could be directly injected for determination with recoveries ranging from 94.5 to 104%.
Acetates↗
First derivative spectrophotometric determination of strychnine and brucine in nux vomica liquid extract.
A simple, rapid, and accurate method is presented for determining strychnine and brucine in nux vomica liquid extract. The 2 alkaloids are directly extracted from the alkaline liquid extract with 0.1N H2SO4, and the liberated alkaloids are determined separately using first derivative spectrophotometry. The minimum strychnine and brucine concentrations detectable by means of this method are 3.0 micrograms/mL. The method has been successfully applied to the analysis of 3 batches of nux vomica liquid extract.
Plant Extracts↗
Reduction of alcohol induced sleep time in albino mice by potentized Nux vomica prepared with 90% ethanol.
UNLABELLED: Male adult albino mice were administered potentized Nux vomica 30 c (Nux v). The drug was mixed with sterile distilled water at 0.05 ml/2 ml water and given at 0.05 ml/individual. Control consisted of blank ethanol solution. Ethanolic extract from the seeds of Strychnos nuxvomica L was mixed with 90% ethanol 1:100 and sonicated for 30 s at 20 KHz. This was further diluted and sonicated in 30 steps to produce Nux v 30 c. Six hours after treatment, mice were given 25% ethanol i.p. at 4 g/kg body wt. The duration of sleep time starting from the loss of righting reflex until its restoration was recorded for each mouse. The duration of sleep time with ethanol was recorded in four sessions for the same group of mice with an interval of 10 d between sessions. TREATMENTS: session 1 with control solution, 2 with Nux v (oral), 3 with control solution and 4 with Nux v (i.p.). Nux v (oral) produced the shortest sleep time as compared to other treatments which did not differ from each other significantly with respect to sleep time. In another experiment Nux v 30 c was prepared with distilled water and pure absolute ethanol by the above process of successive dilution and sonication. These two preparations together with Nux v 30 c, prepared with 90% ethanol, were tested on mice for their effect on alcohol-induced sleep time. Only Nux v 30 c prepared with 90% ethanol was effective in reducing the sleep time in mice. It is concluded that the solution structure of ethanol/water mixture carries the specificity of the Nux v at ultra high dilution. It is further concluded that the effect is mediated through oral receptors.
Animals↗
Cytotoxicities of alkaloids from processed and unprocessed seeds of Strychnos nux-vomica.
AIM: To examine the cytotoxicities of 6 crude Strychnos alkaloid fractions from the seeds of Strychnos nux-vomica unprocessed or processed with various traditional processing methods and 13 pure Strychnos alkaloids from the fractions. METHODS: Using cell culture, their inhibitory effects on Vero cell growth-inhibition assay, and host cell DNA synthesis by [3H]thymidine ([3H]TdR) uptake assay. RESULTS: The IC50 of processed seeds were 155% and 212% of unprocessed ones in cell growth-inhibition assay and in [3H]TdR uptake assay, respectively. The IC50 of 13 compounds were 0.45-0.80 mmol.L-1 and 0.50-12 mmol.L-1, respectively. The processing method with sand bath exhibited a wide safety margin compared with other traditional processing methods or no processing. The isomers of Strychnos alkaloids and their N-oxides showed much lower cytotoxicities among these alkaloids. Isobrucine N-oxide showed the lowest cytotoxicity. The contents of isomers and N-oxides of Strychnos alkaloids were the highest in the sand processing. CONCLUSION: Processing of nux vomica plays a critical role in its toxicity.
Alkaloids↗
The anti-tumor effects of alkaloids from the seeds of Strychnos nux-vomica on HepG2 cells and its possible mechanism.
To screen the anti-tumor effects of the four alkaloids: brucine, strychnine, brucine N-oxide and isostrychnine from the seed of Strychnos nux-vomica, MTT assay was used to examine the growth inhibitory effects of these alkaloids on human hepatoma cell line (HepG2). Brucine, strychnine and isostrychnine revealed significant inhibitory effects against HepG2 cell proliferation, whereas brucine N-oxide didn't have such an effect. In addition, brucine caused HepG2 cell shrinkage, membrane blebbing, apoptotic body formation, all of which are typical characteristics of apoptotic programmed cell death. The results of flow cytometric analysis demonstrated that brucine caused dose-dependent apoptosis of HepG2 cells through cell cycle arrest at G0/G1 phase, thus preventing cells entering S or G2/M phase. Immunoblot results revealed that brucine significantly decreased the protein expression level of cyclooxygenase-2, whereas increased the expression caspase-3 as well as the caspase-3-like protease activity in HepG2 cells, suggesting the involvement of cyclooxygenase-2 and caspase-3 in the pro-apoptotic effects exerted by brucine. Therefore, this paper indicate that the major alkaloids present in the seed of Strychnos nux-vomica are effective against HepG2 cells proliferation, among which brucine proceed HepG2 cells death via apoptosis, probably through the participation of caspase-3 and cyclooxygenase-2.
Alkaloids↗
Potentized Mercuric chloride and Nux vomica facilitate water permeability in erythrocytes of a fresh-water catfish Clarius batrachus under acute ethanol intoxication.
OBJECTIVES: The primary biomolecular target of a homeopathic potency is unknown. If it is a plasma membrane protein such as water-channel protein, the drug would alter water permeation in cells. Therefore, the objective is to see if potentized homeopathic drugs like Mercuric chloride 30c and Nux vomica 30c could alter permeation of water through the erythrocytes of a fresh water fish under acute ethanol intoxication. LOCATION: The work was carried out in the Zoology Laboratory of Visva Bharati University, Santiniketan, West Bengal, India. SUBJECT: Live freshwater catfish. DESIGN: Erythrocytes collected from fish with and without ethanol intoxication were incubated in distilled water at 30 degrees C for 30 minutes with Ethanol 30c (control), Merc cor 30c (test 1), and Nux vomica 30c (test 2). Merc cor 30c and Nux vom 30c were prepared by successive dilution of the respective mother tinctures with 90% ethanol (1:100) followed by sonication at 20 kHz for 30 seconds in 30 steps. Ethanol 30c was prepared in the same way from 90% ethanol diluted with 90% ethanol. In another experiment, fish were pretreated with Ethanol 30c and Nux vom 30c followed by ethanol injection at 2 g/kg of body weight. Then their erythrocytes were tested in vitro with the same potencies. After centrifugation of blood samples, fluid part was removed, erythrocyte pellets dried in a BioChemical Oxygen Demand (BOD; Atlas Surgical, New Delhi, India) incubator at 90 degrees C for 12 hours and intracellular water content measured. RESULTS: Red blood cells (RBCs) from ethanol-injected fish permeated more water than those from normal fish. Water permeation was enhanced with Merc cor 30c and Nux vom 30c. RBCs from fish pretreated with Nux vom 30c imbibed more water in in vitro treatments than those from fish pretreated with Ethanol 30c. CONCLUSION: Because water channel proteins or aquaporins are mainly responsible for water transport through the plasma membrane of RBCs, it is thought that potentized drugs interact with these proteins, thereby facilitating water influx in the cells.
Alcoholic Intoxication↗