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

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

Liver injury in malaria infected patients in Douala-Cameroon and its association with poor medical practice

Abstract Background Malaria is an endemic mosquito-borne disease in sub-Saharan regions, including Cameroon. Due to the obligatory hepatic stage of its pathogenic agents, malaria can induce liver damage if not properly treated. Hence, we assessed the impact of malaria infection on liver transaminases among febrile patients consulting at the Deido District Hospital, Douala-Cameroon, in regard to their attitude towards the practice of preventive measures, treatment, and management of malaria. Methods Over 10 weeks, 150 febrile patients and 28 healthy individuals serving as the control group were enrolled and their blood samples screened for Plasmodium species by Giemsa Staining and liver injury evaluated by measuring the serum level of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities. The socio-demographic characteristics of participants and their attitude towards the practice of preventive measures, treatment, and management of malaria were collected using a structured- questionnaire. Results Among tested febrile patients, 113 (75%) were malaria-positive. Females were more affected (65.5%) than males; the most affected age group were adults between 30-60 years (55.8%). A significant association (p˂0.05; relative risk [RR] = 1.424 or p˂0.05; RR = 1.947) was found between malaria infection and non-use of mosquito nets or insecticides, respectively. The serum level of ALT and AST activities in malaria-positive were significantly (p<0.05) increased, compared to healthy or malaria-negative individuals. Furthermore, transaminase activity was significantly (p<0.05) elevated in non-practitioners of preventive measures; and in patients who engaged in auto-medication or traditional medication, compared to those who sought treatment from health centers. Conclusion Our findings demonstrated that non-practice of preventive measures, improper treatment and management of malaria infection can lead to an abnormal increase in serum level of transaminases which may reflect liver injury

In silico and in vitro screening of licensed antimalarial drugs for repurposing as inhibitors of hepatitis E virus

International audienceHepatitis E virus (HEV) infection is emerging in Cameroon and represents one of the most common causes of acute hepatitis and jaundice. Moreover, earlier reports showed evidence of falciparum malaria/HEVcoexistence. Although the Sofosbuvir/Ribavirin combination was recently proposed in the treatment of HEV-infected patients, no specific antiviral drug has been approved so far, thereby urging the search for new therapies. Fortunately, drug repurposing offers a good alternative to this end. In this study, we report the in silico and in vitro activities of 8 licensed antimalarial drugs and two anti-hepatitis C virus agents used as references (Sofosbuvir, and Ribavirin), for repurposing as antiviral inhibitors against HEV. Compounds were docked against five HEV-specific targets including the Zinc-binding non-structural protein (6NU9), RNA-dependent RNA polymerase (RdRp), cryoEM structure of HEV VLP, genotype 1 (6LAT), capsid protein ORF-2, genotype 3 (2ZTN), and the E2s domain of genotype 1 (3GGQ) using the iGEMDOCK software and their pharmacokinetic profiles and toxicities were predicted using ADMETlab2.0. Their in vitro effects were also assessed on a gt 3 p6Gluc replicon system using the luciferase reporter assay. The docking results showed that Sofosbuvir had the best binding affinities with 6NU9 (− 98.22 kcal/mol), RdRp (− 113.86 kcal/mol), 2ZTN (− 106.96 kcal/mol), while Ribavirin better collided with 6LAT (− 99.33 kcal/mol). Interestingly, Lumefantrine showed the best affinity with 3GGQ (-106.05 kcal/mol). N-desethylamodiaquine and Amodiaquine presented higher binding scores with 6NU9 (− 93.5 and − 89.9 kcal/mol respectively vs − 80.83 kcal/mol), while Lumefantrine had the greatest energies with RdRp (− 102 vs − 84.58), and Pyrimethamine and N-desethylamodiaquine had stronger affinities with 2ZTN compared to Ribavirin (− 105.17 and − 102.65 kcal/mol vs − 96.04 kcal/mol). The biological screening demonstrated a significant (P < 0.001) antiviral effect on replication with 1 µM N-desethylamodiaquine, the major metabolite of Amodiaquine. However, Lumefantrine showed no effect at the tested concentrations (1, 5, and 10 µM). The biocomputational analysis of the pharmacokinetic profile of both drugs revealed a low permeability of Lumefantrine and a specific inactivation by CYP3A2 which might partly contribute to the short half-time of this drug. In conclusion, Amodiaquine and Lumefantrine may be good antimalarial drug candidates for repurposing against HEV. Further in vitro and in vivo experiments are necessary to validate these predictions