| Kinase Assay: |
| Biosci Biotechnol Biochem. 2004 Jan;68(1):126-31. | | Anisodamine causes the changes of structure and function in the transmembrane domain of the Ca(2+)-ATPase from sarcoplasmic reticulum.[Pubmed: 14745174] |
CONCLUSIONS:
The effects of Anisodamine on the Ca(2+)-ATPsae of sarcoplasmic reticulum (SR) were investigated by using differential scanning calorimetry to measure the ability of Anisodamine to denature the transmembrane domain and the cytoplasmic domain. Anisodamine significantly altered the thermotropic phase behaviors of the transmembrane domain of purified Ca(2+)-ATPase. Specifically, the melting temperature of the transmembrane domain moved toward lower temperatures with the concentrations of Anisodamine increasing and the thermotropic phase peak was abolished at 10 mM, indicating that the stabilized structure of the transmembrane domain in the presence of Ca2+ could be destabilized by Anisodamine. Decreases of the intrinsic fluorescence and increases of the extrinsic fluorescence of ANS, a fluorescent probe, showed the exposure of tryptophan and hydrophobic region, respectively, suggesting again that Anisodamine caused a less compact conformation in the transmembrane domain. A marked inhibition of the Ca2+ uptake activity of SR Ca(2+)-ATPase was observed when the addition of Anisodamine. The drug did not affect the cytoplasmic domain of the enzyme and only slightly decreased the ATPase activity of the enzyme at concentrations up to 10 mM. This was likely due to the destabilized protein transmembrane domain.
CONCLUSIONS:
To sum up, our results revealed that Anisodamine interacted specifically with the transmembrane domain of SR Ca(2+)-ATPase and inhibited the Ca2+ uptake activity of the enzyme. |
|
| Animal Research: |
| Crit Care Med. 2009 Feb;37(2):634-41. | | Antishock effect of anisodamine involves a novel pathway for activating alpha7 nicotinic acetylcholine receptor.[Pubmed: 19114896 ] |
METHODS AND RESULTS:
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.
CONCLUSIONS:
These findings demonstrate that the antishock effect of Anisodamine is intimately linked to alpha7nAChR-dependent anti-inflammatory pathway. | | Eur J Pharmacol. 2002 Mar 29;439(1-3):21-5. | | Anisodamine inhibits cardiac contraction and intracellular Ca(2+) transients in isolated adult rat ventricular myocytes.[Pubmed: 11937088] | Increased cardiac workload often leads to serious complications during cardiac surgery such as pericardiopulmonary bypass. Various agents have been applied to lower peripheral resistance and cardiac workload, one of which, Anisodamine, is widely used in Asia. However, the direct action of Anisodamine on cardiac contractile property is essentially unknown.
METHODS AND RESULTS:
This study was designed to examine the influence of Anisodamine on ventricular contractile function at the single cardiac myocyte level. Ventricular myocytes from adult rat hearts were stimulated to contract at 0.5 Hz, and mechanical and intracellular Ca(2+) properties were evaluated using an IonOptix Myocam system. Contractile properties analyzed included peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR(90)), maximal velocity of shortening/relengthening (+/-dL/dt), intracellular Ca(2+) fluorescence intensity change (DeltaFFI) and decay (tau). Anisodamine exhibited a concentration-dependent (10(-12)-10(-6) M) inhibition in PS and DeltaFFI, with maximal inhibitions of 44.7% and 47.2%, respectively. Anisodamine inhibited +/-dL/dt, lowered resting FFI but elicited no effect on TPS/TR(90) and tau. Pretreatment with the nitric oxide synthase (NOS) inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME, 100 microM) abolished the inhibitory effect of Anisodamine in cell shortening. In addition, Anisodamine prevented cholinoceptor agonist carbachol-induced positive cardiac contractile response.
CONCLUSIONS:
This study demonstrated a direct cardiac depressive action of Anisodamine at the myocyte level, which may be related to, at least in part, NO production and cholinoceptor antagonism. | | Am J Chin Med. 2011;39(5):853-66. | | Inhibition of endoplasm reticulum stress by anisodamine protects against myocardial injury after cardiac arrest and resuscitation in rats.[Pubmed: 21905277 ] | Anisodamine is a multi-functional bio-alkaloid with vascular activity. Our previous studies have revealed that Anisodamine protects the heart from ischemia/reperfusion (I/R) injury induced by cardiac arrest (CA) and resuscitation. This study aimed to explore whether the protective effect of Anisodamine is mediated by inhibition of the endoplasmic reticulum stress (ERS) response, which has been demonstrated to implicate in various I/R injuries.
METHODS AND RESULTS:
After 5 min of CA induced by electric stimulation, Wistar rats were randomly selected to receive cardiopulmonary resuscitation (CPR, including chest compression and epinephrine infusion) with or without Anisodamine injection (n = 50/group). Hearts were harvested 24 h after the return of spontaneous circulation (ROSC). Sham-operated animals served as non-ischemic controls (n = 10). The survival rate, cardiomyocyte apoptosis, and the protein expression of ERS markers were detected. Thirty-three of the 50 rats in the Ani + CA/R group were successfully resuscitated, whereas only 18 of the 50 rats in the CA/R group gained ROSC. Survival to 24 h was significantly improved in the Anisodamine treatment group (Ani + CA/R, n = 22/50) compared to the group with standard CPR (CA/R, n = 8/50). Anisodamine markedly decreased the number of apoptotic cardiomyocytes, the protein expression of GRP78, CHOP, and the active form of Caspase3 compared to the CA/R group.
CONCLUSIONS:
Our data suggest that Anisodamine protects against cellular damage in rat hearts after CA and resuscitation, at least in part, by inhibiting myocardial ERS. |
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