(Circulation. 1997;96:1386-1389.)
© 1997 American Heart Association, Inc.
Articles |
Amiodarone Inhibits Production of Tumor Necrosis Factor-
by Human Mononuclear Cells
A Possible Mechanism for its Effect in Heart Failure
From the Department of Cardiovascular Medicine, Kyoto University, Japan.
Correspondence to Akira Matsumori, MD, PhD, Department of Cardiovascular Medicine, Kyoto University, 54 Kawaracho Shogoin, Sakyo-ku, Kyoto 606, Japan. E-mail amat{at}kuhp.kyoto-u.ac.jp
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Abstract |
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 Top Abstract IntroductionMethodsResultsDiscussionReferences |
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Background Recent studies suggest that cytokines such as tumor necrosis factor (TNF)-
and
interleukins (ILs) are capable of modulating
cardiovascular function and that drugs used in the
treatment of heart failure have various modulatory effects on the
production of cytokines. This study was performed to
examine the effects of amiodarone (a drug shown to be
beneficial in some patients suffering from heart failure) versus other
antiarrhythmic agents on the production of cytokines in
vitro.
Methods and Results Human peripheral blood
mononuclear cells (PBMC) were obtained from healthy volunteers. PBMC
were cultured with 0.1, 1, and 10 µmol/L of amiodarone,
quinidine, disopyramide, and lidocaine in the presence of
lipopolysaccharide. After 24 hours' incubation, TNF-,
IL-1ß, and IL-6 were measured in the culture supernatants by an
enzyme-linked immunosorbent assay. TNF- production was
inhibited by amiodarone but stimulated by quinidine in a
concentration-dependent manner. Disopyramide and lidocaine
tended to increase TNF- production. IL-6 production
was decreased by amiodarone in all concentrations but was
increased significantly by disopyramide. Modulation of
IL-1ß production by amiodarone was biphasic and
significantly increased at a concentration of 10 µmol/L.
Conclusions These previously unrecognized immunomodulatory effects of amiodarone may contribute to its beneficial effects in heart failure patients.
Key Words: amiodarone • heart failure • immune system • interleukins
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Introduction |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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With the growing use of amiodarone for the treatment of arrhythmias in patients with CHF, a concomitant improvement was noted in measurements of left ventricular function1 2 and exercise tolerance.2 Although amiodarone did not reduce all-cause or sudden death mortality,3 the substantial reduction in the combined end point of cardiac death plus hospitalization for heart failure in patients with nonischemic disease is noteworthy.3 4 These apparent benefits are in marked contrast to the negative inotropic and hemodynamic responses to other antiarrhythmic agents in this patient population5 6 and, along with a low incidence of proarrhythmic events,7 have made amiodarone the antiarrhythmic agent of choice in the treatment of CHF patients.6 7 8
Recent studies from our laboratory have shown that drugs for heart failure modulate the production of cytokines in various ways9 10 11 and suggest that some of these immunomodulatory effects are relevant to the effects of drugs in heart failure patients. Accordingly, the present study was performed to investigate whether the positive hemodynamic effects of amiodarone might be attributable to a modulation of cytokine production.
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Methods |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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PBMC were obtained from healthy volunteers and collected by Ficoll-paque (Pharmacia) density centrifugation. The cells were washed three times with PBS, resuspended in RPMI 1640 medium (Gibco) supplemented with 10% heat-inactivated fetal calf serum (Gibco), 100 U/mL penicillin, 100 µg/mL streptomycin (Gibco), and 50 µmol/L 2-mercaptoethanol, and cultured at 37°C in a humidified 5% CO2 atmosphere. Amiodarone, quinidine, disopyramide and lidocaine (all from Sigma Chemical Co) were dissolved in 0.1% DMSO and diluted with the medium. This concentration of DMSO did not influence the production of cytokines by PBMC. PBMC (2x106 cells/mL in 24-well plates) were stimulated with 1 µg/mL of LPS, to which 0.1, 1, and 10 µmol/L of each drug was added. After 24 hours' incubation, the supernatants were harvested and stored at -80°C until performance of the cytokine assay. TNF-
, IL-1ß, and IL-6 levels in the
culture supernatants were determined by specific enzyme-linked
immunosorbent assay kits (Otsuka Pharmaceutical Co). Because
significant interindividual variability of cytokine
production by PBMC in response to LPS was observed,
cytokine production from two to five subjects was
measured (2 wells from each). None of the agents affected monocyte
viability, verified by staining with trypan blue.
Statistical Analyses
Because the amount of the cytokines produced was
different in each experiment, the effects of the drugs on
cytokine production was expressed as a percent response
compared with the control wells containing LPS. Statistical
analyses were performed by one-way ANOVA with multiple
comparisons by Fisher's protected least significant difference test.
All values are presented as the mean±SE. A value of
P<.05 was considered statistically significant.
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Results |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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TNF-
production was decreased by amiodarone in
a concentration-dependent manner. Compared with a stimulated control of
2.15±0.57 ng/mL, concentrations of 0.1, 1, and 10
µmol/L (n=10 each) decreased TNF- by 70±15%, 61±13%,
and 54±10%, respectively (P<.05 at 1
µmol/L, P<.01 at 10 µmol/L versus
control; Fig 1
). In contrast, quinidine
increased TNF- production in a concentration-dependent
manner. Compared with a stimulated control of 6.69±3.3 ng/mL,
quinidine concentrations of 0.1, 1, and 10 µmol/L (n=4
each) increased TNF- by 117±10%, 131±7%, and 149±13%,
respectively (P<.05 and P<.01 at 1 and 10
µmol/L, respectively; Fig 1). Compared with a stimulated
control of 0.86±0.07 ng/mL, disopyramide increased
the production of TNF- at all concentrations (162±30%,
151±20%, and 147±33%, respectively; n=4 each). However, these
differences did not reach statistical significance.
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P<.01 vs control.
IL-6 production was significantly inhibited by
amiodarone at all three concentrations (67±6%, 65±6%, and
66±3%; P<.0001) compared with a stimulated control value
of 2.1±0.4 ng/mL (Fig 2).
Conversely, in comparison with a stimulated control of 1.7±0.2
ng/mL, disopyramide increased IL-6
production significantly (120±5%, 113±3%, and 122±5%,
respectively; P<.05) whereas quinidine and lidocaine had no
significant effect (Fig 2).
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P<.0001 vs control.
IL-1ß in the presence of amiodarone was slightly decreased at
concentrations of 0.1 and 1 µmol/L (76±11% and 68±9%,
respectively, versus a stimulated control of 8.6±3.8 ng/mL) but
markedly increased at a concentration of 10 µmol/L
(228±72%; P<.05) (Fig 3).
IL-1ß production was not significantly changed by quinidine,
disopyramide, or lidocaine.
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Discussion |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Several studies have measured an improvement in left ventricular ejection fraction or indices of clinical heart failure with amiodarone therapy.1 2 12 This favorable effect is particularly apparent in patients suffering from nonischemic heart disease.4 Modulation of cardiovascular function by cytokines has been well documented.13 Elevated concentrations of TNF-
have
been reported in patients with chronic heart failure,14 15
and TNF- has been reported to depress myocardial
contractility.16 17 18 19 Furthermore, TNF-,
IL-1ß, and interferon-
have cytotoxic effects on cultured cardiac
myocytes.20 In addition to these humoral effects,
cytokines may activate cytotoxic T-cells, thereby
causing direct injury to the myocyte.21 IL-6 may also
exert a negative inotropic effect,16 and mice
overexpressing both IL-6 and IL-6 receptors have been reported to
develop cardiac hypertrophy.22 Other recent
observations suggest that growth abnormalities, which accompany
hypertrophy of the overloaded myocardium, may
play an important role in the deterioration of the condition of
patients with chronic heart failure.23
Recent studies from our laboratory14 have shown that
increased concentrations of circulating TNF- are found more often in
patients with nonischemic than those with ischemic
heart failure. In our animal models of nonischemic heart
failure induced by viral myocarditis,24 25 plasma TNF-
concentration was increased in the blood, and exogenously given
anti–TNF- antibody improved both myocardial injury and survival of
the animals.26 Furthermore, in the same model, the
expression of TNF- messenger RNA was increased in the acute stage
and persisted into the chronic stage,27 and
immunohistochemical analyses revealed that
endothelial cells and interstitial
macrophages were positive for TNF-. These observations
suggest that TNF- plays an important role in the pathogenesis of
nonischemic heart failure.
The reduced production of TNF- and IL-6 by
amiodarone measured in the present study was in sharp
contrast to the effects produced by three other antiarrhythmic agents
and may explain the hemodynamic effects of
amiodarone noted in clinical studies. Therapeutic plasma
concentrations of amiodarone range between 0.5 and 2
µg/mL, which is equivalent to 0.87 to 3.1
µmol/L, within the range of concentrations used in these
experiments.
The mechanisms by which amiodarone modulates cytokine production remain to be clarified. The involvement of ion channels in T-lymphocyte activation and proliferation is well established.28 29 The involvement of K+ channels in lymphocyte activation and cytotoxic function has been reported recently with the use of the scorpion-derived peptide margatoxin, a specific K+ channel blocker in lymphocytes.30 Vesnarinone, which has been reported to reduce K+ current,31 inhibits the production of cytokines.10 Because amiodarone blocks the inward rectifier K+ channel,32 it may inhibit cytokine release by modulating ion channels. This hypothesis, if verified, may help in the development of more targeted pharmacological treatments of CHF.
Additional studies are necessary to confirm our findings using lymphocytes from patients with CHF. Our observations do not establish a direct connection between the effects of amiodarone on cytokines and its hemodynamic effects. Additional studies will be needed to examine this link in vivo in animal models, as well as by measuring cytokines in patients suffering from CHF who are treated with amiodarone.
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Selected Abbreviations and Acronyms |
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Acknowledgments |
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We would like to thank Y. Ohmoto and H. Toriyama for their assistance in the assay of cytokines and T. Nakano and S. Sakai for preparing the manuscript.
Received April 16, 1997; revision received June 4, 1997; accepted June 10, 1997.
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References |
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