The cytotoxic effects of a traditional Zulu remedy,impila (Callilepis laureola)

Abstract

The traditional Zulu remedy impila (Callilepis laureola) can cause acute fatal hepatocellular necrosis, especially in children. We investigated the mechanism(s) of toxicity using HuH-7 hepatocytes. Impila tubers were extracted with boiling water and the aqueous extract was used at different concentrations to study the effects on the morphology of the cells. Flow cytometry and labelling with fluorescent antibodies to tubulin were also used. At high concentrations, necrosis occurred; however, at lower concentrations, the extracts gave rise to a variety of changes including hypercondensation of chromatin, multinucleate cells, nuclear fragmentation and apoptosis. In addition, we observed destruction of cytoplasmic tubulin. These findings give further insight into the mechanism of toxicity of herbal remedies containing atractyloside.

Keywords

apoptosis Callilepis laureola hepatotoxicity HuH-7 hepatocytes traditional medicine tubulin

References

Maberley J . Joint meeting of the British and South African Associations for the Advancement of Science. 1930.

Seedat Y , Hitchcock P . Acute renal failure from Callilepis laureola . S Afr Med J 1971; 45: 832–833 .

Wainwright J , Schonland M . Toxic hepatitis in black patients in Natal . S Afr Med J 1977; 51: 571–573 .

Bhoola K . A clinico-pathological and biochemical study of the toxicity of Callilepis laureola (Impila) in Medicine, University of Natal, Durban 1983.

Grobler A , Koen M , Brouwers F . Planttoksiene en lewersiektes by kinders in Pretoria en omgewing . Tijdschr Kindergeneeskd 1997; 65: 99–104 .

Brookes K , Candy H , Pegel K . Atractylosides in Callilepis laureola (Asteraceae) . S Afr J Chem 1983; 36: 65–68 .

Watson A , Coovadia H , Bhoola K . The clinical syndrome of Impila (Callilepis laureola) poisoning in children . S Afr Med J 1979; 55: 290–292 .

Neame P , Pillay V . Spontaneous hypoglycaemia, hepatic and renal necrosis following the intake of herbal medicines . S Afr Med J 1964; 38: 729–732 .

Obatomi D , Bach P . Atractyloside nephrotoxicity: in vitro studies with suspensions of rat renal fragments and precision-cut cortical slices . In Vitr Mol Toxicol 2000; 13: 25–36 .

10.

Popat A , Shear N , Malkiewicz I , et al. The toxicity of Callilepis laureola, a South African traditional herbal medicine . Clin Biochem 2001; 34: 229–236 .

11.

Allman D , Harris R , Green D . Site of action of atractyloside in mitochondria. II. Inhibition of oxidative phosphorylation . Arch Biochem Biophys 1967; 122: 766–782 .

12.

Steenkamp V , Stewart M , Zuckerman M . Detection of poisoning by Impila (Callilepis laureola) in a mother and child . Hum Exp Toxicol 1999; 18: 594–597 .

13.

Steenkamp V , Stewart MJ , van der Merwe S , Zuckerman M , Crowther NJ . The effect of Senecio latifolius a plant used as a South African traditional medicine, on a human hepatoma cell line . J Ethnopharmacol 2001; 78: 51–58 .

14.

Sherwood S , Schimke R . Cell cycle analysis of apoptosis using flow cytometry . Methods Cell Biol 1995; 46: 77–79 .

15.

Kiernan J . Histological and histochemical methods. London: Pergamon Press , 1990.

16.

Davidson AM , Halestrap AP . Liver mitochondrial pyrophosphate concentration is increased by ca2+ and regulates the intramitochondrial volume and adenine nucleotide content . Biochem J 1987; 246: 715–723 .

17.

Petit P , Goulbern M , Diolez P , Susin S , Zamzami N , Kroemer G . Disruption of the outer mitochondrial membrane as a result of large amplitude swelling: the impact of irreversible permeability transition . FEBS Lett 1998; 426: 111–116 .

18.

Muller M , Strand S , Hug H , et al. Drug induced apoptosis in hepatoma cells is mediated by the CD95(APO1/Fas) receptor ligand system and involves activation of wild type p53 . J Clin Invest 1997; 99: 403–413 .

19.

Hsu I , Tokiwa T , Bennett W , et al. p53 gene mutation and integrated hepatitis B viral DNA sequences in human liver cancer cell lines . Carcinogenesis 1993; 14: 987–992 .

20.

Obatomi D , Bach P . Inhibition of mitochondrial respiration and oxygen uptake in isolated rat renal tubular fragments by atractyloside . Toxicol Lett 1996; 89: 155–161 .

21.

Huang Y , Chou C . Bcl-2 blocks apoptotic signal of transforming growth factor-beta in human hepatoma cells . J Biomed Sci 1998; 5: 185–191 .

22.

Schteingart C , Pomilio A . Atractyloside, toxic compound from Wedelia glauca . J Nat Prod (United States) 1984; 47: 1046–1047 .

23.

Georgiou M , Siandidou L , Hatzis T , Papadatos J , Koutselinis A . Hepatotoxicity due to Atractylis gummifera L . Clin Toxicol 1988; 26: 487–493 .

24.

Roeder E , Bourauel T , Meier U , Wiedenfeld H . Diterpenoid glycosides from Iphiona aucheri . Phytochemistry 1994; 37: 353–355 .

25.

Obatomi D , Blackburn R , Bach P . Effects of the calcium channel blocker verapamil and sulphydryl reducing agent dithiothreitol on atractyloside toxicity in precision-cut rat renal cortical and liver slices . Food Chem Toxicol 2001; 39: 1013–1021 .