Propolis prevents hepatorenal injury induced by chronic exposure to carbon tetrachloride

Abstract

Carbon tetrachloride (CCl(4)) is a well-known hepatotoxicant, and its exposure induces hepatorenal injury via oxidative stress and biochemical alterations. This study had been conducted to confirm the protective role of propolis extract on CCl(4)-induced hepatorenal oxidative stress and resultant injury. Propolis extracts collected from Gwalior district and 24 female Sprague Dawley rats were used for experiment. Animals were exposed to CCl(4) (0.15 mL/kg, i.p.) for 12 weeks (5 days/week) followed by treatment with propolis extract (200 mg/kg, p.o.) for consecutive 2 weeks. CCl(4) exposure significantly depleted blood sugar and hemoglobin level and raised the level of transaminases, alkaline phosphatase, lactate dehydrogenase, protein, urea, albumin, bilirubin, creatinine, triglycerides, and cholesterol in serum. Lipid peroxidation was enhanced, whereas GSH was decreased significantly in liver and kidney in CCl(4)-intoxicated group. Ethanolic extract of propolis successfully prevented these alterations in experimental animals. Activities of catalase, adenosine triphosphatase, glucose-6-phosphatase, acid, and alkaline phosphatase were also maintained towards normal with propolis therapy. Light microscopical studies showed considerable protection in liver and kidney with propolis treatment, thus, substantiated biochemical observations. This study confirmed hepatoprotective potential of propolis extract against chronic injury induced by CCl(4) by regulating antioxidative defense activities.

Figure 1

Showing therapeutic potential of propolis on CCl₄-induced alteration in liver marker enzymes of serum. Values are mean ± SE of n = 6 in each group. @Significant for ANOVA at 5% level. P value CCl₄ versus normal at #≤0.05; P value treatment versus CCl₄ at ∗ ≤ 0.05 for Student's t-test; F variance for ANOVA of AST = 60.3^†, ALT = 145^†, LDH = 114^†, SALP = 204^†. Abbreviations: Control (C); Carbon tetrachloride (CCl₄); Propolis (P); Silymarin (S); Aspartate aminotransferase (AST); Alanine aminotransferase (ALT); Lactate dehydrogenase (LDH), and Alkaline phosphatase (SALP).

Figure 2

Showing therapeutic potential of propolis on CCl₄-induced alteration in serological parameters. Values are mean ± SE of n = 6 in each group. @Significant for ANOVA at 5% level. P value CCl₄ versus normal at #≤0.05; P value treatment versus CCl₄ at ∗ ≤ 0.05 for Student's t-test; F variance for ANOVA of hemoglobin = 4.44^†, blood sugar = 6.57^†, triglycerides = 46.9^†, and cholesterol = 7.81^†. Abbreviations: Control (C); Carbon tetrachloride (CCl₄); Propolis (P); Silymarin (S).

Figure 3

Showing therapeutic potential of propolis on CCl₄-induced alteration in specific serum markers. Values are mean ± SE of n = 6 in each group. @Significant for ANOVA at 5% level. P value CCl₄ versus normal at #≤0.05; P value treatment versus CCl₄ at ∗ ≤ 0.05 for Student's t-test; F variance for ANOVA of albumin = 7.34^†, bilirubin = 39.0^†, urea = 64.1^†, and creatinine = 56.1^†. Abbreviations: Control (C); Carbon tetrachloride (CCl₄); Propolis (P); Silymarin (S).

Figure 4

Showing therapeutic potential of propolis on CCl₄-induced hepatorenal oxidative stress. Values are mean ± SE of n = 6 in each group. @Significant for ANOVA at 5% level. P value CCl₄ versus normal at #≤0.05; P value treatment versus CCl₄ at ∗ ≤ 0.05 for Student's t-test; F variance for ANOVA of hepatic LPO = 104^†, renal LPO = 45.2^†, hepatic GSH = 50.3^†, renal GSH = 27.7^†, hepatic CAT = 37.7^†, and renal CAT = 18.9^†. Abbreviations: Control (C); Carbon tetrachloride (CCl₄); Propolis (P); Silymarin (S).

Figure 5

(a) Photomicrograph of control liver with polygonal hepatocytes cordially arranged around hepatic central vein. Hepatic nuclei (N), uniform sinusoidal spaces (S) terminating into central vein (CV) were clearly visible (200x). (b) Liver sections of CCl₄ induced chronic hepatic injury with disturbed cord arrangement of hepatocytes, vacuolation (V) due to collapsed cellular membranes, high accumulation of nonparenchymal or inflammatory cells (NP) and irregular sinusoids (200x). (c and d) Liver sections of CCl₄ induced chronic hepatic injury with disturbed cord arrangement of hepatocytes and high degree of vacuolation (V) due to collapsed cellular membranes (400x). (e) Photomicrograph of liver after propolis therapy with minimal degree of vacuolation, better formed polygonal hepatocytes cordially arranged towards hepatic central vein (CV). Hepatic nuclei (N), uniform sinusoids (S) terminating into central vein were clearly visible (200x). (f) Silymarin therapy with better formed features of liver histology as comparable to experimental control (400x).

Figure 6

(a) Photomicrograph of kidney of control rat showing well-formed Bowman's capsule and normal glomeruli (G), uniform space between glomerulus and capsule wall, tubular lumen with basal and apical nuclei (200x). (b) CCl₄ administration caused swelling in glomerulus and tubular obstruction. loss of glomerulus space (thick arrows) and (200x). (c) CCl₄ exposure caused swelling in glomerulus so that space between glomerulus and capsule wall (curved arrow) was reduced (400x). (d): Administration of propolis retained the kidney histoarchitecture with opened lumen of tubules (T) and uniform space between glomerulus and capsule wall (curved arrow) (200x). (e) Propolis therapy maintained uniform space between glomerulus and capsule wall (curved arrow) (400x). (f) Silymarin therapy recovered the kidney histoarchitecture with wider lumen of tubules (T) (400x).