Artemether-Lumefantrine treatment combined with albendazole and ivermectin induced genotoxicity and hepatotoxicity through oxidative stress in Wistar rats

Peer review under responsibility of Mansoura University.

Figures & data

Table 1 Experimental animal grouping and drug dosage and administration.

Experimental groups	Drug doses (mg/kg bwt)	Exposure durations
A	Distilled water	0.5 ml for 3 consecutive days
B	20/120 AL	0.5 ml for 3 consecutive days
C	10/60 AL	0.5 ml for 3 consecutive days
D	40/240 AL	0.5 ml for 3 consecutive days
E	20/120 AL + 400 ABZ	0.5 ml AL + 0.5 ml ABZ for 3 days
F	10/60 AL + 200 ABZ	0.5 ml AL + 0.5 ml ABZ for 3 days
G	40/240 AL + 800 ABZ	0.5 ml AL + 0.5 ml ABZ for 3 days
H	20/120 AL + 3 IVR	0.5 ml AL + 0.5 ml IVR for 3 days
I	10/60 AL + 1.5 IVR	0.5 ml AL + 0.5 ml IVR for 3 days
J	40/240 AL + 6 IVR	0.5 ml AL + 0.5 ml IVR for 3 days
K	20/120 AL + 400 ABZ	0.5 ml AL for 3days + 0.5 ml ABZ only on the 3rd day
L	20/120 AL + 3 IVR	0.5 ml AL for 3days + 0.5 ml ABZ only on the 3rd day
M	40 CYP	0.5 ml for 3 consecutive days

AL = Artemether-Lumefantrine (Artemisinin-based combination therapy; Coartem^®); IVR = Ivermectin; ABZ = Albendazole; CYP = Cyclophosphamide (positive control). Drugs were constituted using distilled water (vehicular solvent; negative control).

Fig. 1 Abscess on the thigh region of the right leg of rat exposed to Artemether/Lumefantrine (20/120 mg/bwt) and Ivermetin (3 mg/kg), sign of clinical toxicity to drug exposure.

Fig. 2 a. Effects of drug exposure on serum ALT activity of rat. End point represents mean ± SE for 5 rats. Values are significantly different; ^ap < 0.05; ^bp < 0.01; ^cp < 0.001 and *p > 0.05 compared to negative control. b. Effects of drug exposure on serum AST activity of rat. End point represents mean ± SE for 5 rats. Values are significantly different; ^ap < 0.05; ^bp < 0.01; ^cp < 0.001 and *p > 0.05 compared to negative control. c. Effects of drug exposure on total bilirubin concentration of rat. End point represents mean ± SE for 5 rats. Values are significantly different; ^ap < 0.05; ^bp < 0.01; ^cp < 0.001 and *p > 0.05 compared to negative control.

Table 2 Effects of drug combinations on serum CAT, SOD and MDA in treated and control rats.

Treatment	SOD	(95% CI)	CAT	(95% CI)	MDA	(95% CI)
A	31.29 ± 2.43	(27.32–35.26)	1.98 ± 0.02	(1.36–2.60)	28.64 ± 1.16	(25.42–31.86)
B	29.73 ± 1.68*	(27.49–31.98)	1.86 ± 0.13*	(1.22–2.49)	32.72 ± 2.82*	(27.66–37.77)
C	29.72 ± 3.49*	(25.57–33.87)	1.94 ± 0.09*	(1.41–2.48)	29.62 ± 1.66*	(25.01–34.23)
D	23.63 ± 3.32^b	(19.96–27.30)	1.75 ± 0.17*	(1.28–2.22)	41.80 ± 0.96^c	(36.37–35.88)
E	22.86 ± 0.93^c	(18.18–23.53)	1.35 ± 0.21^a	(1.04–1.66)	38.60 ± 0.93^b	(33.24–43.95)
F	26.56 ± 0.95^a	(21.16–31.97)	1.66 ± 0.01^a	(1.36–1.95)	40.41 ± 1.49^c	(33.50–47.31)
G	20.65 ± 2.02^c	(17.81–23.48)	1.03 ± 0.08^b	(0.26–1.80)	49.68 ± 2.00^c	(44.14–55.22)
H	24.63 ± 1.45^a	(23.59–31.67)	1.54 ± 0.16^a	(1.09–1.99)	29.25 ± 3.08*	(26.26–32.24)
I	30.43 ± 0.68*	(25.76–35.10)	1.75 ± 0.26*	(1.30–2.20)	30.67 ± 1.13*	(24.75–36.59)
J	22.18 ± 1.61^b	(17.75–26.88)	1.36 ± 0.01^a	(1.04–1.68)	42.49 ± 0.38^c	(35.88–49.10)
K	26.57 ± 1.71^a	(21.77–31.28)	1.63 ± 0.20^a	(1.07–2.19)	32.68 ± 2.74^a	(27.86–33.24)
L	25.53 ± 1.16^a	(24.31–30.75)	1.84 ± 0.03*	(1.47–2.21)	31.79 ± 1.53*	(27.54–36.04)
M	25.48 ± 2.36^a	(21.70–29.25)	1.42 ± 0.13^a	(1.06–1.79)	35.72 ± 3.03^a	(30.09–41.34)

End point represents mean ± SE for 5 rats. Values are significantly different; ^ap < 0.05; ^bp < 0.01; ^cp < 0.001 and *p > 0.05 compared to negative control. SOD (superoxide dismutase; U/mg protein), CAT (catalase; μmol/mg protein), MDA (malondialdehyde; nmol/ml), (95% confidential interval).

Fig. 3 Frequency of micronucleated polychromatic erythrocytes in bone marrow cells of rats exposed to drugs and controls. End point represents mean ± SE for 5 rats. Values are significantly different; ^ap < 0.05; ^bp < 0.01; ^cp < 0.001 and *p > 0.05 compared to negative control.

Fig. 4 (a–b): Asterisks showed normal polychromatic erythrocytes while arrows showed micronucleated polychromatic erythrocytes from bone marrow cells of rats.

Fig. 5 Liver tissues of rats exposed to drug combinations (H&E, x 400). (a) Section of liver from rat in the negative control group showing relatively normal hexagonal or pentagonal lobule of hepatocytes. (b) Single cell hepatocellular necrosis and kupffer cell hyperplasia (c) Multiple foci vacuolar changes and congestion of the sinusoids. (d) Thinning of hepatic cord and single cell hepatocellar necrosis. (e) Vacuolar changes in the hepatocytes and mild congestion of the sinusoids. (f) Severe congestion of the sinusoids by inflammatory cells.