Interaction of agonists of a different subtype of the nAChR and carvacrol with GABA in Ascaris suum somatic muscle contractions

Abstract

Resistance of parasitic nematodes to anthelmintic drugs is a growing problem in human and veterinary medicine. The molecular mechanisms by which nematodes become resistant are different, but certainly one of the possible processes involves changing the drug binding site on the specific receptor. The significance of changes in individual subtypes of nicotinic acetylcholine receptors (nAChRs) for the development of resistance has not been clarified in detail. This study investigates the interaction of antinematodal drugs, agonist of different types of nAChRs and carvacrol with gamma aminobutyric acid (GABA) on the contractions of parasitic nematode A. suum. In our study, GABA (3 μM) produced significant increase of contractile EC₅₀ value for pyrantel, and nonsignificant for bephenium and morantel, from 8.44 to 28.11 nM, 0.62 to 0.96 µM, and 3.72 to 5.69 nM, respectively. On the other hand, the maximal contractile effect (R _max) did not change in the presence of GABA. However, when A. summ muscle flaps were incubated with GABA 3 μM and carvacrol 100 μM, the EC₅₀ value of pyrantel, bephenium, and morantel was increased significantly to 44.62 nM, 1.40 μM, and nonsignificantly to 7.94 nM, respectively. Furthermore, R _max decreased by 70, 60, and 65%. Presented results indicate that the combined use of GABA receptor agonists and nicotinic receptor antagonists can effectively inhibit the neuromuscular system of nematodes, even when one of the nicotinic receptor subtypes is dysfunctional, due to the potential development of resistance.

Keywords: Carvacrol; GABA; Interaction; Resistance; nAChR.

Figure 1:

A Original recording of isometric contractions of A. suum muscle flap induced by increasing concentrations of pyrantel and the effect of GABA (3 μM) and Carvacrol (100 μM) on those contractions; B the concentration-response plot for pyrantel control (n = 6, blue), in the presence of GABA 3 μM (n = 6, green), in the presence of GABA and Carvacrol 100 μM (n = 6, red) and after washing (n = 6, dashed black) (mean ± S.E.).

Figure 2:

A Original recording of isometric contractions of A. suum muscle flap induced by increasing concentrations of bephenium and the effect of GABA (3 μM) and Carvacrol (100 μM) on those contractions; B the concentration-response plot for bephenium control (n = 6, blue), in the presence of GABA 3 μM (n = 6, green), in the presence of GABA and Carvacrol 100 μM (n = 6, red) and after washing (n = 6, dashed black) (mean ± S.E.).

Figure 3:

A Original recording of isometric contractions of A. suum muscle flap induced by increasing concentrations of morantel and the effect of GABA (3 μM) and carvacrol (100 μM) on those contractions; B the concentration-response plot for morantel control (n = 6, blue), in the presence of GABA 3 μM (n = 6, green), in the presence of GABA and carvacrol 100 μM (n = 6, red) and after washing (n = 6, dashed black) (mean ± S.E.).