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Abstract
A non-steroidal anti-inflammatory drug (NSAID) has many adverse effects including cardiovascular (CV) risk. Diclofenac among the nonselective NSAIDs has the highest CV risk such as congestive heart failure, which resulted commonly from the impaired cardiac pumping due to a disrupted excitation-contraction (E-C) coupling. We investigated the effects of diclofenac on the L-type calcium channels which are essential to the E-C coupling at the level of single ventricular myocytes isolated from neonatal rat heart, using the whole-cell voltage-clamp technique. Only diclofenac of three NSAIDs, including naproxen and ibuprofen, significantly reduced inward whole cell currents. At concentrations higher than 3 μM, diclofenac inhibited reversibly the Na+ current and did irreversibly the L-type Ca2+ channels-mediated inward current (IC50=12.89±0.43 μM) in a dose-dependent manner. However, nifedipine, a well-known L-type channel blocker, effectively inhibited the L-type Ca2+ currents but not the Na+ current. Our finding may explain that diclofenac causes the CV risk by the inhibition of L-type Ca2+ channel, leading to the impairment of E-C coupling in cardiac myocytes.
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Fig. 1.
Inhibition of the Na+- and the Ca2+-sensitive inward current by three NSAIDs. (A) Representative currents before and after application of the drugs denoted above the corresponding trace. The drugs were applied at a concentration of 10 μM each. The amplitudes of the initial transient component and the slowly decayed components were measured at positions marked by closed () and open circles (❍), respectively. (B) Summary of the normalized data for the effect of drugs on two components. Relative inhibition (%) for the Na+-sensitive initial transient (black bar) and the nifedipine-sensitive slowly decayed components (open bar) are shown with number of observations. Data were normalized to currents measured before application of each drug.
Fig. 2.
Representative traces of whole-cell currents elicited by step depolarizations in single cardiac myocytes. (A) Inhibition of the inward current induced by diclofenac. Changes in whole-cell currents evoked at –40 and 0 mV from a holding potential of –100 mV in bath solution containing 140 mM Na+ before (left) and after adding diclofenac (middle), and following washout (right), respectively. Dotted lines in A and B indicate the zero current level. (B) Currents induced by depolarization as indicated above the traces, in Na+-free bath solution before and after the addition of diclofenac. (C) Current-voltage relationship measured from the peak current of the traces in panel B. Diclofenac of 100 μM was applied. Outward components were not detected due to the presence of Ba2+, instead of Ca2+ in the bath.
Fig. 3.
Inhibition of the Na+ and the Ba2+ components by diclofenac. (A) Representative currents inhibited by drugs denoted above the right trace. With the application of diclofenac (100 μM), nifedipine (1 μM), or nickel (300 μM), reduced currents were shown on the right. The amplitudes of the initial transient component and the slowly decayed components were measured at the positions marked by the closed circle () and triangle (▴), respectively. (B) Summary of the normalized data for the effect of drugs on the two components. Relative inhibitions (%) of the Na+-sensitive initial transient and the nifedipine-sensitive components are shown in upper and lower panel with number of observations, respectively. Data were normalized to currents measured before application of each drug. Scale bars are equal to 1 nA and 50 ms.
Fig. 4.
Dose-dependent inhibition of the L-type current by diclofenac. (A) Dose-response relationship of the inhibitory effect of diclofenac on peak L-type (IBa) currents in cardiomyocytes, with the numbers of cells. The molar concentration of diclofenac is given. (B) Representative traces of L-type currents reduced by diclofenac. Step depolarizations were applied from HP of –50 mV to +60 mV in 10 mV increments.
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