Comparative assessment of hepatoprotective properties of Artesunate and flavonoids from Artemisia annua on acetaminophen and carbon tetrachloride-induced cytotoxicity in primary mice hepatocytes

Marie Ange Djeungoue Petga; Arnaud Fondjo Kouam; Rosine Désirée Chougouo Kengne; Boris Rosnay Galani Tietcheu; Josué Simo Louokdom; Claude Bérenger Ngantchouko Ngalemo; Pascal Dieudonné Chuisseu Djamen; Paul Fewou Moundipa

doi:10.1016/j.metop.2023.100241

Comparative assessment of hepatoprotective properties of Artesunate and flavonoids from Artemisia annua on acetaminophen and carbon tetrachloride-induced cytotoxicity in primary mice hepatocytes

Metabol Open. 2023 Apr 6:18:100241. doi: 10.1016/j.metop.2023.100241. eCollection 2023 Jun.

Authors

Marie Ange Djeungoue Petga¹, Arnaud Fondjo Kouam^{2

3}, Rosine Désirée Chougouo Kengne¹, Boris Rosnay Galani Tietcheu^{3

4}, Josué Simo Louokdom¹, Claude Bérenger Ngantchouko Ngalemo¹, Pascal Dieudonné Chuisseu Djamen^{1

3}, Paul Fewou Moundipa³

Affiliations

¹ Higher Institute of Health Sciences, Université des Montagnes, P.O. Box 208, Bangangté, Cameroon.
² Medical Research and Applied Biochemistry Laboratory, Department of Biomedical Sciences, Faculty of Health Sciences, University of Buea, PO Box 63, Buea, Cameroon.
³ Laboratory of Pharmacology and Toxicology, Department of Biochemistry, Faculty of Science, University of Yaoundé 1, P.O. Box 812, Yaounde 1, Cameroon.
⁴ Laboratory of Applied Biochemistry, Department of Biological Sciences, Faculty of Science, University of Ngaoundere, PO Box 454, Ngaoundere, Cameroon.

Abstract

Background: Artesunate (ART) is a semi-synthetized molecule from Artemisinin, an active compound isolated from the medicinal plant Artemisia annua, widely used for the treatment of malaria. Previous studies reported that ART may exert a dual effect on the liver. Accordingly, this study investigated the potential protective action of ART against Acetaminophen (APAP) and Carbon tetrachloride (CCl₄)-induced hepatotoxicity in primary mice hepatocytes, in comparison to that of flavonoid extracted from A. annua (FAA). In addition, the antioxidant properties of FAA were also assessed.

Methods: The antioxidant activities of FAA and Ascorbic acid (ASC) (0.01-100 μg/mL) were assessed through inhibition of lipid peroxidation, reduction of ferric and phosphomolydenum, and hydroxyl and DPPH radicals scavenging assays. The hepatoprotective effects of FAA and ART (0.1-100 μg/mL) were evaluated against APAP (11 mM) or CCl4 (4 mM) induced oxidative damage in primary mouse hepatocytes. Biochemical parameters associated with hepatotoxicity assessed include cell viability, cell membrane integrity, cellular glutathione, and antioxidant enzyme activities.

Results: The obtained finding revealed FAA displayed a remarkable antioxidant activities as evidenced by the low IC₅₀/EC₅₀ values (3.85-19.32 μg/mL), comparable to that of ASC (3.26-18.04 μg/mL). When tested at 10 μg/mL, both FAA and ART significantly (p˂0.05) preserved cell viability, inhibited alanine aminotransferase leakage and lipid membrane peroxidation, and restored superoxide dismutase and catalase activities and glutathione content induced by APAP or CCl₄ in a similar way as Silymarin. However, ART showed a significant (p˂0.05) cytotoxic effect on hepatocytes at 100 and 1000 μg/mL and did not confer obvious protection at 100 μg/mL.

Conclusion: Overall, our data demonstrated that ART harms mice hepatocytes at high concentration while conferring relative protection against APAP and CCl₄-hepatotoxicity at low concentration. In contrast, FAA effectively protects liver cells without cytotoxicity effect, event at 100 μg/mL. Accordingly, ART should be given to the patient only under a medical prescription.

Keywords: Acetaminophen; Antioxidant activities; Artesunate; Carbon tetrachloride; Flavonoid from Artemesia annua; Hepatotoxicity.