Doctoral thesis on kava

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Doctoral thesis on kava

Doctoral Thesis On Kava

In this thesis the hepatotoxic properties of the two phytomedicines kava kava and cimicifuga racemosa have been investigated. This topic is a prevailing problem as herbal medicines are quite popular today and not much is known about their toxicological profile.

Especially hepatotoxicity is a wide-spread problem of medicines in general as most of them are metabolized in the liver and the liver therefore represents a near target.

Ingestion of kava has been associated with liver damage and therefore Kava has been withdrawn from the market in different countries.

For cimicifuga so far not much evidence for liver toxicity in humans exists. However, lately it was found that rats fed with high doses of cimicifuga developed microvesicular liver steatosis.

In order to examine the in vitro toxicity of the two herbs the hepatocarcinoma cell line HepG2, the hepatoblastoma cell line HUH6, and isolated rat liver mitochondria have been used.

Three kava extracts a methanolic and an acetonic root extract; a methanolic leaf extract and an ethanolic cimicifuga extract have Doctoral thesis on kava investigated.

In the first project chapter 6 toxicity of kava kava was investigated. It was found that all three kava extracts showed concentration-dependent toxicity in the general cytotoxicity tests. Experiments in mitochondria revealed that the kava extracts inhibited and uncoupled root extracts or only uncoupled leaf extract the respiratory chain and decreased the mitochondrial membrane potential; in addition, beta-oxidation was inhibited.

Furthermore, oxidized glutathione was slightly increased and ATP content of the cells was maintained. Apoptosis was found in HepG2 cells for all three kava extracts. These findings of mitochondria-related toxicity affected respiratory chain, decreased mitochondrial membrane potential and increased reactive oxygen species ROS production might result in the opening of the permeability transition pore and consequently in the rupture of the outer mitochondrial membrane, thus possibly leading to the release of cytochrome c and subsequently to apoptosis.

These events might contribute to kava associated hepatotoxicity, especially in predisposed patients having mitochondrial damages. In the second project chapter 7 two methods for cell death determination were applied and compared in the hepatoma cell lines HepG2 and HUH6 utilizing kava as death-inducing agent: These results on the one hand confirm that kava indeed induces cell death; on the other hand they show that the GFPmethod can also be employed in liver cell lines.

Therefore, the GFP-method is suited as an easy, reliable, cost-effective method to screen substances for their hepatotoxic potential.

In a further study chapter 8 the toxicity profile of the phytomedicine cimicifuga was assessed. High doses of cimicifuga extract caused microvesicular liver steatosis in rats.

Doctoral thesis on kava

Based on these findings in vitro experiments were performed to further investigate the hepatotoxic potential of this plant. General cytotoxicity tests revealed concentration-dependent toxicity.

In mitochondria, cimicifuga extract was able to inhibit b-oxidation, to uncouple the respiratory chain and to reduce the mitochondrial membrane potential. In addition, HepG2 cells underwent apoptosis when incubated with cimicifuga.

Doctoral thesis on kava

It might therefore be assumed that the impairment of the respiratory chain causes a decrease of the mitochondrial membrane potential, which in turn could lead to the opening of the permeability transition pore and to apoptosis. Whereas the inhibition of the b-oxidation may lead to accumulation of fatty acids and subsequently to liver steatosis.

Toxicity was discovered at concentrations higher than the ones expected in humans.

Doctoral thesis on kava

It is therefore conjectured, that hepatotoxicity in humans only occurs under certain conditions. Finally, it can be concluded that although kava kava and cimicifuga racemosa displayed clear toxicity in the in vitro situation it has to be further investigated whether these toxicities can be extrapolated to humans.

Factors which have to be taken into account when estimating the toxic potential for humans include the concentrations used in the in vitro tests in comparison to portal venous concentrations in humans after intake of the extracts, the metabolism in the body, predisposing factors for liver toxicity like age, gender and pre-existing liver diseases, and polymedication.

Nevertheless, these cytotoxicity tests represent a valuable tool for evaluating the toxic potential of herbal products before they reach the market. Of course, other experiments animal experiments, for example would have to be added, if an assessment corresponding to the rules of the regulatory offices is sought.

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Courses can be selected from those offered by Hanken or by other universities or by National Doctoral Programmes, such as KATAJA and KAVA (FDPE) in Finland or within the EIASM program abroad. The courses do not have to be directly focused on the specific theme of the thesis.

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