Inhibition of mouse liver respiration by Chelidonium majus isoquinoline alkaloids

doi:10.1016/j.toxlet.2003.09.007

Toxicology Letters

Volume 146, Issue 1, 15 December 2003, Pages 37-47

https://doi.org/10.1016/j.toxlet.2003.09.007 Get rights and content

Abstract

The alkaloids from Chelidonium majus L. which had a significant inhibitory effect in mitochondrial respiration were those which contain a positive charge due to a quaternary nitrogen atom, i.e., chelerythrine, sanguinarine, berberine and coptisine, both with malate+glutamate or with succinate as substrates. When malate+glutamate was used as substrate, chelerythrine and berberine, which contain methoxy groups, were particularly more active, since they had a strong effect even at low concentrations. In submitochondrial particles, berberine and coptisine had a marked inhibitory effect on NADH dehydrogenase activity but practically no effect on succinate dehydrogenase activity, whereas chelerythrine and sanguinarine inhibited more strongly succinate dehydrogenase than NADH dehydrogenase, which is in agreement with the results found for mitochondrial respiration. Protopine and allocryptopine, which did not inhibit mitochondrial respiration, strongly inhibited NADH dehydrogenase in submitochondrial particles, but had no effect on succinate dehydrogenase activity.

Introduction

Chelidonium majus L. is a plant which grows in the wild in Southern and Central Europe, part of Asia, North America and in the Azores archipelago (Kadan et al., 1990, Pavao and Pinto, 1995, Colombo and Bosisio, 1996). Its use as a medicinal plant is very ancient (Paris and Moyse, 1967, Duke, 1985, Xeme Pharmacopee Francaise, 1989, Bezanger-Beauquesne et al., 1990). The medicinal properties mentioned above can be ascribed to the more than 27 alkaloids present in the root and aerial part of the plant, which belong to three main groups (Fig. 1): (i) benzo[c]phenanthridines, with two subgroups, (i.a) quaternary, like chelerythrine and sanguinarine or (i.b) tertiary, like chelidonine, (ii) protopine and derived thereof, such as protopine and allocryptopine, and (iii) protoberberines such as berberine and coptisine (Lavenir and Paris, 1965, Taborska et al., 1994, Pavao and Pinto, 1995, Tome and Colombo, 1995, Colombo and Bosisio, 1996). The interest of this plant for medicinal purposes implies the need to know as much as possible about the effects on metabolic processes of the alkaloids it contains. Several alkaloids with the same or related structures have been found to interfere with respiration, either at the level of the electron transport chain (Schewe and Müller, 1976) or as uncouplers (Vallejos and Rizzotto, 1972). Since mitochondrial respiration is the core of metabolic energy, and therefore a process with major importance, in the present work we investigated the effect of some of these alkaloids (Fig. 1) in respiration-linked processes. We selected alkaloids from each of the main groups found in the plants collected in S. Miguel Island, Azores (Pavão and Pinto, 1995). The effects of phenanthrene were also monitored, to allow for effects due only to the aromatic structure of the molecules.

The aim of the present work is (a) to ascertain whether the effects detected follow a similar pattern within each group; (b) if any effect which occurs on oxygen uptake can be explained by events at the level of NADH dehydrogenase (NADH:ubiquinone oxidoreductase, EC 1.6.99.3) or succinate dehydrogenase (succinate:ubiquinone oxidoreductase, EC 1.3.99.1). These two complexes were chosen by their crucial role in the respiratory chain and by evidence from other authors that these systems might be affected by compounds of this type (Schewe and Müller, 1976; McNaught et al., 1995, McNaught et al., 1996).

Section snippets

Animals

The animals used were male albino mice, with approximately 12 weeks of age and an average weight of 20–25 g. The animals were fed ad libitum with a commercial chow and tap water.

Alkaloids

Chelidonine, berberine chloride and sanguinarine chloride were purchased from Sigma. The other alkaloids were a kind gift from Prof. Slavik (Masaryk University, Brno, Czech Republic). The alkaloids and phenanthrene were used in methanolic solutions. The effect of methanol was tested for all types of experiment, in the

Oxygen uptake by intact mitochondria

The effects of the several groups of alkaloids on oxygen uptake in mitochondria showed different patterns. Chelerythrine and sanguinarine, both containing a quaternary nitrogen atom with a methyl group, strongly inhibited succinate-dependent respiration and, to a lesser extent, malate–glutamate respiration, while chelidonine, an uncharged phenanthridine derivative, had virtually no effect (Fig. 2). Protopine and allocryptopine, both uncharged and with a CO group, also had no apparent effect (

Discussion

The alkaloids with a charge due to a quaternary nitrogen atom presented a high inhibitory activity on oxygen uptake (Fig. 2, Fig. 4). Some authors have already observed that alkaloids containing a quaternary nitrogen atom are the ones with the highest biological activity (Ulrichova et al., 1984, Dostal and Potacek, 1990; McNaught et al., 1995, McNaught et al., 1996).

The alkaloids which contain a methyl group linked to the quaternary nitrogen atom seemed to have a more significant effect on

Acknowledgements

We would like to thank Professor Slavik (Purkine University, Brno, Czech Republic) for the kind gift of alkaloids from Chelidonium majus.

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Inhibition of mouse liver respiration by Chelidonium majus isoquinoline alkaloids

Abstract

Introduction

Section snippets

Animals

Alkaloids

Oxygen uptake by intact mitochondria

Discussion

Acknowledgements

Anal. Biochem.

FEBS Lett.

Pharmacol. Res.

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochem. Pharmacol.

Biochem. Pharmacol.

Tibtech.

J. Biol. Chem.

Arch. Biochem. Biophys.

Toxicol. Lett.

FEBS Lett.