OBJECTIVE: To explore the synergic mechanism of ginsenoside Rg₁ (Rg₁) and aconitine (AC) by acting on normal neonatal rat cardiomyocytes (NRCMs) and pentobarbital sodium (PS)-induced damaged NRCMs. METHODS: The toxic, non-toxic, and effective doses of AC and the most suitable compatibility concentration of Rg₁ for both normal and damaged NRCMs exposed for 1 h were filtered out by 3- (4,5)-dimethylthiahiazo (-z-y1)-3,5-diphenytetrazoliumromide, respectively. Then, normal NRCMs or impaired NRCMs were treated with chosen concentrations of AC alone or in combination with Rg₁ for 1 h, and the cellular activity, cellular ultrastructure, apoptosis, leakage of acid phosphatase (ACP) and lactate dehydrogenase (LDH), intracellular sodium ions [Na⁺], potassium ions [K⁺] and calcium ions [Ca²⁺] levels, and Nav1.5, Kv4.2, and RyR₂ genes expressions in each group were examined. RESULTS: For normal NRCMs, 3000 µ mol/L AC significantly inhibited cell viability (P<0.01), promoted cell apoptosis, and damaged cell structures (P<0.05), while other doses of AC lower than 3000 µ mol/L and the combinations of AC and Rg₁ had little toxicity on NRCMs. Compared with AC acting on NRCMs alone, the co-treatment of 3000 and 10 µ mol/L AC with 1 µ mol/L Rg₁ significantly decreased the level of intracellular Ca²⁺ (P<0.01 or P<0.05), and the co-treatment of 3000 µ mol/L AC with 1 µ mol/L Rg₁ significantly decreased the level of intracellular Ca²⁺ via regulating Nav1.5, RyR₂ expression (P<0.01). For damaged NRCMs, 1500 µ mol/L AC aggravated cell damage (P<0.01), and 0.1 and 0.001 µ mol/L AC showed moderate protective effect. Compared with AC used alone, the co-treatment of Rg₁ with AC reduced the cell damage, 0.1 µ mol/L AC with 1 µ mol/L Rg₁ significantly inhibited the level of intracellular Na⁺ (P<0.05), 1500 µ mol/L AC with 1 µ mol/L Rg₁ significantly inhibited the level of intracellular K⁺ (P<0.01) via regulating Nav1.5, Kv4.2, RyR₂ expressions in impaired NRCMs. CONCLUSION Rg₁ inhibited the cardiotoxicity and enhanced the cardiotonic effect of AC via regulating the ion channels pathway of [Na⁺], [K⁺], and [Ca²⁺].
Aconitine/pharmacology* ; Animals ; Apoptosis ; Cardiotonic Agents/pharmacology* ; Cardiotoxicity/drug therapy* ; Cell Survival ; Ginsenosides/pharmacology* ; Rats

Quantitative evaluation of analgesic efficacy improves understanding of the antinociceptive mechanisms of new analgesics and provides important guidance for their development. Lappaconitine (LA), a potent analgesic drug extracted from the root of natural Aconitum species, has been clinically used for years because of its effective analgesic and non-addictive properties. However, being limited to ethological experiments, previous studies have mainly investigated the analgesic effect of LA at the behavioral level, and the associated antinociceptive mechanisms are still unclear. In this study, electrocorticogram (ECoG) technology was used to investigate the analgesic effects of two homologous derivatives of LA, Lappaconitine hydrobromide (LAH) and Lappaconitine trifluoroacetate (LAF), on Sprague-Dawley rats subjected to nociceptive laser stimuli, and to further explore their antinociceptive mechanisms. We found that both LAH and LAF were effective in reducing pain, as manifested in the remarkable reduction of nocifensive behaviors and laser-evoked potentials (LEPs) amplitudes (N2 and P2 waves, and gamma-band oscillations), and significantly prolonged latencies of the LEP-N2/P2. These changes in LEPs reflect the similar antinociceptive mechanism of LAF and LAH, i.e., inhibition of the fast signaling pathways. In addition, there were no changes in the auditory-evoked potential (AEP-N1 component) before and after LAF or LAH treatment, suggesting that neither drug had a central anesthetic effect. Importantly, compared with LAH, LAF was superior in its effects on the magnitudes of gamma-band oscillations and the resting-state spectra, which may be associated with their differences in the octanol/water partition coefficient, degree of dissociation, toxicity, and glycine receptor regulation. Altogether, jointly applying nociceptive laser stimuli and ECoG recordings in rats, we provide solid neural evidence for the analgesic efficacy and antinociceptive mechanisms of derivatives of LA.
Aconitine/pharmacology* ; Analgesics/pharmacology* ; Animals ; Pharmaceutical Preparations ; Rats ; Rats, Sprague-Dawley

Chinese patent medicine containing aconitine is the key in clinical rational drug use. These drugs contain Chuanwu, Caowu or Fuzi, and Aconitum brachypodum with functions of expelling wind-dampness or tonifying Yang, all of which shall be used by strictly following the indications and dosage. However, there are many kinds of such drugs. Not only the unfamiliar knowledge of some Chinese and Western physicians about the characteristics of them, but also the combination of multiple drugs from different clinical departments, would increase the risk of aconitine poisoning. Based on the previous research, this paper proposed three core elements "syndrome differentiation-dosage differentiation-toxicity differentiation" from the prescription review and pharmacy consulting work, and objective and standardized evaluation was used to build a risk assessment scale containing 3 categories, 9 items and 36 indicators with Hulisan Jiaonang and Qufeng Zhitong Jiaonang as the example. This scale was used to evaluate the risk of a therapeutic regimen before and after the implementation. According to the verification of the existing adverse reaction cases, the risk assessment scale can be used to indicate the risk of drug treatment program and identify the risk level of drug treatment status. This paper tried to provide a methodological paradigm for scientific and objective evaluation on the safety of Chinese patent medicines, and help to identify the key links and risk prevention in the rational use by Chinese medicine physicians and pharmacists.
Aconitine ; adverse effects ; pharmacology ; Aconitum ; chemistry ; Drugs, Chinese Herbal ; adverse effects ; pharmacology ; Humans ; Nonprescription Drugs ; adverse effects ; pharmacology ; Risk Assessment

In order to investigate the effects of aconitine on [Ca2+] oscillation patterns in cultured myocytes of neonatal rats, fluorescent Ca2+ indicator Fluo-4 NW and laser scanning confocal microscope (LSCM) were used to detect the real-time changes of [Ca2+] oscillation patterns in the cultured myocytes before and after aconitine (1.0 micromol/L) incubation or antiarrhythmic peptide (AAP) and aconitine co-incubation. The results showed under control conditions, [Ca2+] oscillations were irregular but relatively stable, occasionally accompanied by small calcium sparks. After incubation of the cultures with aconitine, high frequency [Ca2+] oscillations emerged in both nuclear and cytoplasmic regions, whereas typical calcium sparks disappeared and the average [Ca2+] in the cytoplasm of the cardiomyocyte did not change significantly. In AAP-treated cultures, intracellular [Ca2+] oscillation also changed, with periodic frequency, increased amplitudes and prolonged duration of calcium sparks. These patterns were not altered significantly by subsequent aconitine incubation. The basal value of [Ca2+] in nuclear region was higher than that in the cytoplasmic region. In the presence or absence of drugs, the [Ca2+] oscillated synchronously in both the nuclear and cytoplasmic regions of the same cardiomyocyte. It was concluded that although oscillating strenuously at high frequency, the average [Ca2+] in the cytoplasm of cardiomyocyte did not change significantly after aconitine incubation, compared to the controls. The observations indicate that aconitine induces the changes in [Ca2+] oscillation frequency other than the Ca2+ overload.
Aconitine/*pharmacology ; Animals, Newborn ; Calcium Signaling/*drug effects ; Cells, Cultured ; Myocytes, Cardiac/cytology ; Myocytes, Cardiac/*metabolism ; Rats, Sprague-Dawley

Ectopic spontaneous activity originated from the injured dorsal root ganglion (DRG) neurons in rats was recorded through single dorsal root fiber. The firing patterns induced by veratridine and aconitine, inhibitors of inactivation gate of sodium channel operating on different binding sites, were compared. In the same neuron, veratridine (1.5 approximately 5.0 micromol/L) caused slow wave oscillations of interspike intervals (ISIs), while aconitine (10 approximately 200 micromol/L) caused tonic firing. Moreover, even if the background firing patterns were various and the reagent concentrations used were different, veratridine and aconitine still induced slow wave oscillations and tonic firing patterns, respectively. The results suggest that veratridine and aconitine induce different firing patterns in injured DRG neurons, which may relate to their inhibitory effects on different binding sites of the sodium channel.
Aconitine ; pharmacology ; Animals ; Electrophysiological Phenomena ; physiology ; Female ; Ganglia, Spinal ; injuries ; physiopathology ; Male ; Neurons ; pathology ; physiology ; Rats ; Rats, Sprague-Dawley ; Sodium Channel Agonists ; Sodium Channels ; physiology ; Veratridine ; pharmacology

Aconitine ; analogs & derivatives ; pharmacology ; Analgesics, Non-Narcotic ; pharmacology ; Animals ; Female ; Intestine, Small ; physiology ; Male ; Mice ; Muscle Contraction ; drug effects ; Muscle, Smooth ; physiology ; Rabbits

AIMTo clarify mechanisms that the antiarrhythmic effects of matrine and berbamine are weaker than those of amiodarone and RP58866.

METHODSExperimental arrhythmic models were induced by aconitine, coronary artery ligation and electric stimulation in rats and rabbits. Whole-cell patch-clamp techniques were used to record IK1, IKr, IKs and Ito.

RESULTSMatrine and berbamine significantly increased the dose of aconitine for induction of ventricular premature and ventricular tachycardia in rats, decreased the number of arrhythmias induced by coronary artery ligation in rats and increased ventricular fibrillation threshold (VFT) induced by electric stimulation in rabbits, but the anti-arrhythmic potency of matrine and berbamine was lower than that of amiodarone and RP58866. The inhibitory actions of matrine and berbamine on IK1, IKr, IKs, Ito were lower than those of amiodarone and RP58866. The IC50 of matrine for IK1, IKr, IKs, Ito were (46 +/- 3), (32.9 +/- 1.2), (37 +/- 8) and (7.6 +/- 0.5) mol x L(-1), respectively. The IC50 of amiodarone for IK1, IKr, IKs, Ito were (21 +/- 5) , (3.7 +/- 0.7), (5.9 +/- 0.9) and (5.9 +/- 0.6) mol x L(-1), respectively.

CONCLUSIONThe inhibitory actions of matrine and berbamine on IK1, IKr, IKs, Ito were lower than those of amiodarone and RP58866, which might be the reason that the antiarrhythmic effects of matrine and berbamine were weaker than those of amiodarone and RP58866.

Aconitine ; Alkaloids ; pharmacology ; Amiodarone ; pharmacology ; Animals ; Anti-Arrhythmia Agents ; pharmacology ; Arrhythmias, Cardiac ; chemically induced ; physiopathology ; prevention & control ; Benzylisoquinolines ; pharmacology ; Chromans ; pharmacology ; Dogs ; Female ; Guinea Pigs ; Male ; Piperidines ; pharmacology ; Potassium Channels ; drug effects ; Quinolizines ; Rabbits ; Rats

OBJECTIVETo study the correlation between content changes of Diester diterpenoid alkaloids (DDAs) content and safety of the processed Fuzi.

METHODSequential and Bliss methods were used to evaluate the safety of 7 kinds of Fuzi processed with different processing time. The relationship between ED50, TD50, TI and content changes of DDAs of those processed Fuzi was studied, the correlation between the content changes and effect of different processed Fuzi was analyzed, and the toxicity of those processed Fuzi with multiple linear regression was tested.

RESULTFuzi with good efficiency and safety contains proper hypaconitine (HA) and mesaconitine (MA). Aconitine (AC) interfered efficacy of Fuzi (negative correlation), HA showed positive correlation with toxicity and efficacy of Fuzi.

CONCLUSIONHA and MA kept in determinate proportion are very important for the safety and effectivity of processed Fuzi.

Aconitine ; analogs & derivatives ; metabolism ; Alkaloids ; chemistry ; Animals ; Diterpenes ; chemistry ; metabolism ; Drug-Related Side Effects and Adverse Reactions ; Drugs, Chinese Herbal ; adverse effects ; metabolism ; pharmacology ; Magnoliopsida ; chemistry ; Male ; Rats ; Time Factors

OBJECTIVETo investigate the ability of the pericardium meridian (PM) to mitigate or enhance the cardiotoxic effects of aconitine injected at specific acupoint and non-acupoint sites in rabbits.

METHODSThis study consisted of 3 experiments that were designed to test the effects of injection of 30 μg/kg of aconitine at acupoints on the PM (Test 1), at non-acupoint sites on the PM (Test 2), and at acupoints on other meridians and non-meridian sites (Test 3). In Test 1, 24 rabbits were randomly assigned to receive injections at Quze (PC3), Tianquan (PC2), or intramuscularly. In Test 2, 24 rabbits were randomly assigned to receive injections of aconitine at non-acupoint I, non-acupoint II, or intramuscularly. In Test 3, 48 rabbits were randomly assigned to receive injections at Neiguan (PC6), Sanyinjiao (SP6), Yangjiao (GB35), a non-meridian and non-acupoint site (NMNA), intravenously, and intramuscularly. Electrocardiographs of the rabbits were performed before, during and after injection to determine the incidence of arrhythmia, latency of ventricular rhythm, and recovery rate after aconitine injection. The recovery time index and extent of arrhythmia scores were calculated.

RESULTSIn all groups the incidence of arrhythmia was 100%, and the latency of ventricular rhythm was less than 30 min. In Tests 1 and 2, the recovery rates of the Quze and non-acupoint II groups were significantly higher than those of the muscular group (P < 0.05). In Test 3, the recovery time index and extent of arrhythmia scores of the Neiguan group were low. There were no significant differences between the other acupoint groups, or the NMNA group, when compared with the group receiving aconitine intramuscularly.

CONCLUSIONSAcupoints or non-acupoints along the PM could reduce the severity of the arrhythmia induced by aconitine in healthy rabbits. Meridians play an important role in protecting body functions.

Aconitine ; adverse effects ; pharmacology ; Acupuncture Points ; Acupuncture Therapy ; methods ; Analysis of Variance ; Animals ; Arrhythmias, Cardiac ; chemically induced ; diagnosis ; Disease Models, Animal ; Electrocardiography ; Male ; Meridians ; Pericardium ; drug effects ; Rabbits ; Random Allocation

This paper was aimed to investigate the inhibitory effect of aconitine(AC) on angiotensin Ⅱ(Ang Ⅱ)-induced H9 c2 cell hypertrophy and explore its mechanism of action. The model of hypertrophy was induced by Ang Ⅱ(1×10-6 mol·L-1),and cardiomyocytes were incubated with different concentrations of AC. Western blot was used to quantify the protein expression levels of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),β-myosin heavy chain(β-MHC),and α-smooth muscle actin(α-SMA). Real-time quantitative PCR(qRT-PCR) was used to quantify the mRNA expression levels of cardiac hypertrophic markers ANP,BNP and β-MHC. In addition,the fluorescence intensity of the F-actin marker,an important component of myofibrils,was detected by using laser confocal microscope. AC could significantly reverse the increase of total protein content in H9 c2 cells induced by Ang Ⅱ; qRT-PCR results showed that AC could significantly inhibit the ANP,BNP and β-MHC mRNA up-regulation induced by AngⅡ. Western blot results showed that AC could significantly inhibit the ANP,BNP and β-MHC protein up-regulation induced by AngⅡ. In addition,F-actin expression induced by Ang Ⅱ could be inhibited by AC,and multiple indicators of cardiomyocyte hypertrophy induced by Ang Ⅱ could be down-regulated,indicating that AC may inhibit cardiac hypertrophy by inhibiting the expression of hypertrophic factors,providing new clues for exploring the cardiovascular protection of AC.
Aconitine ; pharmacology ; Actins ; metabolism ; Angiotensin II ; Atrial Natriuretic Factor ; metabolism ; Cardiac Myosins ; metabolism ; Cardiomegaly ; Cells, Cultured ; Humans ; Hypertrophy ; Myocytes, Cardiac ; drug effects ; Myosin Heavy Chains ; metabolism ; Natriuretic Peptide, Brain ; metabolism