Coenzyme Q10 nullified khat-induced hepatotoxicity, nephrotoxicity and inflammation in a mouse model

Abstract

Ethnopharmacological relevance: The consumption of khat (Catha Edulis, Forsk) is on the rise despite the much publicized associated deleterious health effects. How chemicals present in khat, affect various physiological and biochemical processes requires further scrutiny. A clear understanding of these processes will provide an avenue for countering khat-driven negative effects using appropriate pharmacological and/or nutritional interventions.

Aim of the study: The current study investigated the effect of khat on vital physiological and biochemical processes such as oxidative stress, inflammation and immune responses and the role of Coenzyme-Q₁₀ (CoQ₁₀), a potent antioxidant and anti-inflammatory, in modulating any negative effects due to khat exposure.

Methodology: Three (3) weeks old forty (40) Swiss albino mice were randomly assigned into four treatment groups (n = 10). The first group was the control that was not administered with khat or CoQ₁₀. The second group received 200 mg/kg body weight (b/w) of CoQ₁₀, while the third group received 1500 mg/kg b/w of khat extract and finally the forth group was co-treated with 200 mg/kg b/w of CoQ₁₀ and 1500 mg/kg b/w of khat extract. The experiment was conducted for 90 days after which samples were collected for physiological and biochemical analyses.

Results: The effects of khat and CoQ₁₀ on the weights of brain, liver, kidney and spleen was determined. Administration of khat decreased the levels of RBCs and its subtypes (MCV, MCH, RDW-SD and RDW-CV), a clear indicator of khat-induced normochromic microcytic anemia. White blood cells (lymphocytes, monocytes, neutrophils and eosinophil) which are vital in responding to infections were markedly elevated by khat. Moreover, these results provide evidence for khat-induced liver and kidney injury as shown by increased biomarkers; AST, ALT, GGT and creatinine respectively. Standard histopathological analysis confirmed this finding for khat-driven liver and kidney injury. Further studies showed evidence for khat-induced inflammation and oxidative stress as depicted by increased levels of the pro-inflammatory cytokine TNF-alpha and elevation of GSH in the brain, liver and spleen. Remarkably, this is the first study to demonstrate the potential of CoQ₁₀ in ameliorating khat-induced negative effects as outlined. CoQ₁₀ supplementation restored the khat-induced reduction in RBC subtypes, and was protective against liver and kidney injury as shown by the appropriate biomarkers and standard histopathology analysis. The other significant finding was the CoQ₁₀-driven normalization of GSH and TNF-α levels, indicating a protective effect from khat-driven oxidative stress and inflammation respectively.

Conclusion: From this study, we conclude that CoQ₁₀ may be useful in nullifying khat-driven deleterious events among chronic khat users.

Keywords: Alternative medicine; Biochemistry; Biological sciences; Coenzyme-Q10; Health sciences; Immune system; Inflammation; Khat toxicity; Khat-induced anemia; Neuroscience; Oxidative stress; Pharmaceutical science; Toxicology.

Figure 1

Comparison of the effects of Khat and Coenzyme Q₁₀ administration on organ weight (g) in male Swiss albino mice. The figures shows the change in organ weight of the brain (Figure A), liver (Figure B) and spleen (Figure C). Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Comparisons between various groups were done by one way ANOVA with Tukey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001∗∗∗P ≤ 0.0001). n = 10. Bars represent mean ± SEM.

Figure 2

Mean number of RBC (Figure A), HCT (Figure C) and HGB (Figure C) from blood of male Swiss albino mice. Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. The levels of each RBC subtype were compared by One way ANOVA with Turkey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001∗∗∗P ≤ 0.0001). n = 10. Bars represent mean ± SEM. u/L: microliters per Liter, g/dL: grams per deciliter.

Figure 3

Mean comparison of red blood cell indices following CoQ₁₀ and khat administration in male Swiss albino mice. The figures shows MCV (mean corpuscular volume) (Figure A), MCH (mean corpuscular hemoglobin) (Figure B), RDW-SD (red cell distribution width –standard deviation) (Figure C) and RDW-CV (red cell distribution width –coefficient of variation) (Figure D). Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Red Blood Cell subtype levels from each group were compared by One way ANOVA with Tukey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001∗∗∗P ≤ 0.0001). n = 10. Bars represent mean ± SEM.

Figure 4

Comparison of the effect of khat and or CoQ₁₀ supplementation on the WBC in male Swiss albino mice. Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Comparisons between various groups were done using one way ANOVA with Tukey multiple comparisons post hoc test. Bars represent mean ± SEM.

Figure 5

Effect of khat and or CoQ₁₀ supplementation on WBC subtypes in male Swiss albino mice. Figure A: Effect on lymphocytes, B: Monocytes, C: Neutrophils, D: Eosinophils and E: Basophils. Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Comparisons were done with one way ANOVA with Tukey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001 ∗∗∗P ≤ 0.0001). uL: microliters. n = 10. Bars represent mean ± SEM.

Figure 6

Comparison of the effect of khat and or CoQ₁₀ supplementation on the platelets in male Swiss albino mice. Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Comparisons between various groups were done by one way ANOVA with Tukey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001∗∗∗P ≤ 0.0001). n = 10 mice per group). Bars represent mean ± SEM. uL: Microliters.

Figure 7

Comparison of the effect of khat and or CoQ₁₀ supplementation on platelet subtypes in male Swiss albino mice. Figure A: Effect on PCT (plateletcrit), B: P-LCR (platelet large cell ratio), C: MPV (mean platelet volume) and D: PDW (platelet distribution width). Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. The platelet subtype levels from each group were compared by one way ANOVA with Tukey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001∗∗∗P ≤ 0.0001) n = 10. Bars represent mean ± SEM.

Figure 8

Effects of CoQ₁₀ and Khat supplementation on the liver transaminases enzymes in male Swiss albino mice. Figure 8A–D shows changes in the activity of A: AST (asparte aminotransferase), B: ALT (Alanine aminotransferase), C: Ratio AST: ALT and D: GGT (Gamma-glutamyl transferase) respectively. Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Comparisons between various groups were done by one way ANOVA with Tukey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤0.001∗∗∗P ≤ 0.0001). n = 10. U/L: Units per liter. Bars represent mean ± SEM.

Figure 9

Comparison of the effect of CoQ₁₀ and Khat on the serum levels of bilirubin and creatinine in male Swiss albino mice. The figure shows the effect of khat and/or CoQ₁₀ on total bilirubin (Figure A) and creatinine (Figure B). Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Comparisons between the various groups were done by one way ANOVA with Tukey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001; ∗∗∗P ≤ 0.0001). n = 10. Bars represent mean ± SEM. umol/L: micromole per liter.

Figure 10

Comparison of the effect of khat and or CoQ₁₀ supplementation on cellular GSH concentration in male Swiss albino mice. Coenzyme-Q10 supplementation modulated khat-induced oxidative stress in the brain (Figure A), liver (Figure B) and spleen (Figure C). Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Comparisons in GSH levels in the brain, liver and kidney from each group was done by one way ANOVA with Turkey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001; ∗∗∗P ≤ 0.0001). Bars represent mean ± SEM. GSH units expressed in micromoles/g (umol/g).

Figure 11

The effect of CoQ₁₀ and Khat supplementation on the levels of the cytokines Tumor Necrosis Factor-alpha (TNF-α) (Figure A), Interferon gamma (IFN-γ) (Figure B) and Interleukin-10 (IL-10) (Figure C). Units pg/mL: Picograms per milliliter. Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Comparisons between the various groups were done by one way ANOVA with Tukey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001; ∗∗∗P ≤ 0.0001). n = 10. Bars represent mean ± SEM.

Figure 12

Comparison of the effect of CoQ₁₀ and Khat supplementation on the levels of the ratios of cytokines Tumor Necrosis Factor-alpha (TNF-α): Interleukin-10 (IL-10) (Figure A) and Interferon gamma (IFN-γ): Interleukin-10 (IL-10) (Figure B). Wild type naïve (Control) group, CoQ₁₀ only administered group, Khat only administered group and Khat-CoQ₁₀ co-administered group. Comparisons between various groups were done by one way ANOVA with Tukey multiple comparisons post hoc test. (Indicated level of significance: ∗P ≤ 0.05; ∗∗P ≤ 0.001; ∗∗∗P ≤ 0.0001). n = 10. Bars represent mean ± SEM.

Figure 13

Comparison of the effect of khat and/or CoQ₁₀ on the liver tissue in male Swiss albino mice. Mice orally administered with Coenzyme Q10 show mild liver damage as shown. (a) Normal liver section from control group, (b) Liver section from mice that received 1500 mg/kg bwt of Khat orally showing focal hepatocyte necrosis (arrow) and hepatocyte cytoplasm vacuolation (star) (x100, H&E). (c)Normal Liver from mice treated with 200 mg/kg of CoQ₁₀ and 1500 mg/kg of body weight of khat (×100, H&E). The results clearly demonstrate that khat induced liver damage; and CoQ₁₀ protected from such injury.

Figure 14

Assessment of the effect of khat and/or CoQ₁₀ on the kidney tissue in male Swiss albino mice. (a) Normal Kidney of mice from control group (b) Focal areas of tubular necrosis in mice administered with Khat administered (arrows) and (c) Normal kidney of ice treated with 200 mg/kg of CoQ₁₀ and 1500 mg/kg of body weight of Khat. (Magnification: × 100, H&E). The results clearly show that khat extract induced kidney injury; and CoQ₁₀ provided some protection from such damage.