aconitine and anisodamine

Vagus nerve stimulation inhibits proinflammatory cytokine production by signaling through the alpha7 nicotinic acetylcholine receptor (alpha7nAChR). Anisodamine, a muscarinic acetylcholine receptor antagonist, has been used clinically in China for treatment of various shocks, but the mechanism was poorly understood. Here, we tested the hypothesis whether anisodamine attained its antishock effect through activation of alpha7nAChR.. : Randomized and controlled in vitro and in vivo study.. Research laboratory and animal facility rooms.. Sprague-Dawley rats, Kunming mice, alpha7nAChR-deficient mice, and RAW264.7 cells.. Sprague-Dawley rats were injected with lipopolysaccharide (LPS) (15 mg/kg, intravenous) to induce septic shock. Methyllycaconitine, a selective alpha7nAChR antagonist, was administered (10 mg/kg, intraperitoneal) 10 minutes before anisodamine (10 mg/kg, intravenous). Mean arterial pressure was monitored and cytokines were analyzed 2 hours after the onset of LPS. In vagotomized mice and alpha7nAChR-deficient mice, the antishock effect of anisodamine was appraised, respectively. RAW264.7 cells were stained by fluorescein isothiocyanate- labeled-alpha-bungarotoxin and the fluorescence intensity was observed. Mice peritoneal macrophages were pretreated and stimulated with LPS, and tumor necrosis factor (TNF)-alpha in the supernatant was measured by enzyme-linked immunosorbent assay.. Methyllycaconitine significantly antagonized the beneficial effect of anisodamine on mean arterial pressure and TNF-alpha, interleukin-1beta expression in response to LPS. The antishock effects of anisodamine were markedly attenuated in vagotomized mice and alpha7nAChR-deficient mice. In vitro, anisodamine significantly augmented the effect of acetylcholine on fluorescence intensity stained with fluorescein isothiocyanate-labeled-alpha-bungarotoxin and TNF-alpha production stimulated with LPS.. These findings demonstrate that the antishock effect of anisodamine is intimately linked to alpha7nAChR-dependent anti-inflammatory pathway.

Topics: Aconitine; alpha7 Nicotinic Acetylcholine Receptor; Animals; Base Sequence; Cell Line; China; DNA Primers; Interleukin-1beta; Macrophages, Peritoneal; Mice; Nicotinic Agonists; Nicotinic Antagonists; Polymerase Chain Reaction; Rats; Rats, Sprague-Dawley; Receptors, Nicotinic; Shock, Septic; Solanaceous Alkaloids; Tumor Necrosis Factor-alpha; Vagotomy

We employed CE to identify mixtures of the toxic alkaloids lappaconitine, bullatine A, atropine sulfate, atropine methobromide, scopolamine hydrobromide, anisodamine hydrobromide, brucine, strychnine, quinine sulfate, and chloroquine in human blood and urine, using procaine hydrochloride as an internal standard. The separation employed a fused-silica capillary of 75 microm id x 60 cm length (effective length: 50.2 cm) and a buffer containing 100 mM phosphate and 5% ACN (pH 4.0). The sample was injected in a pressure mode and the separation was performed at a voltage of 16 kV and a temperature of 25 degrees C. The compounds were detected by UV absorbance at wavelengths of 195 and 235 nm. All the ten alkaloids were separated within 16 min. The method was validated with regard to precision (RSD), accuracy, sensitivity, linear range, LOD, and LOQ. In blood and urine samples, the detection limits were 5-40 ng/mL and linear calibration curves were obtained over the range of 0.02-10 microg/mL. The precision of intra- and interday measurements was less than 15%. Electrophoretic peaks could be identified either by the relative migration time or by their UV spectrum.

Topics: Aconitine; Alkaloids; Atropine; Atropine Derivatives; Electrophoresis, Capillary; Scopolamine; Solanaceous Alkaloids; Strychnine

Anisodamine (Ani) is an alkaloid first isolated in China from the root of Anisodus tanguticus. Ani 10, 15 mg.kg-1 i.v. markedly shortened the duration of arrhythmias induced by aconitine (10 micrograms.kg-1 i.v.) or by BaCl2 (2 mg.kg-1 i.v.) in anesthetized rats. Ani significantly effected on arrhythmias induced by early coronary artery ligation in rats by reducing total numbers of ectopic beats and shortening the duration of ventricular tachycardia and ventricular fibrillation 30 min after coronary ligation. The incidence of ventricular fibrillation of mice induced by chloroform were reduced from 100% to 20% and 10% by i.v. Ani 1 and 10 mg.kg-1 respectively. Ani 0.05, 0.25 mumol.L-1 prolonged the duration of neurologic refractory period of isolated guinea pig left atria and 10 mg.kg-1 i.v. had no effect on tachycardia induced by i.v. isoproterenol (0.01 mg.kg-1) but blocked the stimulation of nervi vagus. After i.v. Ani 15 mg.kg-1 P-P, P-R and QT-c intervals on the ECG were prolonged. Mean arterial pressure and diastolic pressure were reduced but systolic pressure was not effected.

Topics: Aconitine; Animals; Anti-Arrhythmia Agents; Arrhythmias, Cardiac; Autonomic Nervous System; Barium; Barium Compounds; Chlorides; Female; Guinea Pigs; In Vitro Techniques; Male; Mice; Rats; Refractory Period, Electrophysiological; Solanaceous Alkaloids