Genistein Improves Liver Damage in Male Mice Exposed to Morphine

Abstract

Background: Morphine is commonly used to treat severe pain. This substance is significantly metabolized in the liver and causes disturbing effects. Genistein is an isoflavone and has antioxidant properties. The aim of this study was to evaluate the effects of genistein against morphine damages on mouse liver.

Methods: Between May 2017 and March 2018, 48 male mice were divided into six groups (n = 8 in each group). Various doses of genistein (25 and 50 mg/kg) and morphine plus genistein (25 and 50 mg/kg) were administered intraperitoneally to 48 male mice for 20 consequent days. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), serum nitric oxide (NO) levels, liver weight, and the diameter of hepatocytes and central hepatic vein were studied and compared using one-way analysis of variance.

Results: Morphine administration significantly increased the mean diameter of the central hepatic vein (22.76 ± 1.9 μm vs. 15.04 ± 0.60 μm, χ² = 21.814, P = 0.001) and hepatocytes (3.03 ± 0.10 μm vs. 1.10 ± 0.05 μm, χ² = 9.873, P = 0.001) respectively, blood serum NO level (38.00% ± 2.09% vs. 18.72% ± 4.40%, χ² = 20.404, P < 0.001), liver enzyme level (AST: 111.80 ± 5.10 ng/ml vs. 81.93 ± 2.20 ng/ml, χ² = 32.201, P < 0.0001; ALT: 45.14 ± 4.10 ng/ml vs. 35.49 ± 2.50 ng/ml, χ² = 18.203, P < 0.0001; and ALP: 3.28 ± 0.20 ng/ml vs. 2.14 ± 0.10, χ² = 5.04, P < 0.0001, respectively), and decreased liver weight (18.50 ± 0.90 g vs. 27.15 ± 0.50 g, χ² = 22.415, P = 0.001) compared to saline group (0.535-0.750, P < 0.0001). However, administration of genistein plus morphine significantly enhanced liver weight (25 mg/kg: 21.15 ± 2.13 g vs. 18.50 ± 0.90 g, χ² = 19.251, P < 0.0001; 50 mg/kg: 21.20 ± 1.00 g vs. 18.5 ± 0.9 g, χ² = 19.502, P < 0.0001, respectively) and reduced the mean diameter of hepatocyte (25 mg/kg: 2.17 ± 0.30 μm vs. 3.03 ± 0.10 μm, χ² = 22.780, P = 0.001; 50 mg/kg: 2.01 ± 0.20 μm vs. 3.03 ± 0.10 μm χ² = 7.120, P = 0.001, respectively), central hepatic vein (25 mg/kg: 19.53 ± 1.00 μm vs. 22.76 ± 1.90 μm, χ² = 20.681, P = 0.001; 50 mg/kg: 19.44 ± 1.20 μm vs. 22.76 ± 1.90 μm, χ² = 18.451, P = 0.001, respectively), AST (25 mg/kg: 95.40 ± 5.20 ng/ml vs. 111.80 ± 5.010 ng/ml, P < 0.0001; 50 mg/kg: 90.78 ± 6.00 ng/ml vs. 111.80 ± 5.10 ng/ml, χ² = 17.112, P < 0.0001, respectively), ALT (25 mg/kg: 35.78 ± 5.01 ng/ml vs. 45.14 ± 4.10 ng/ml, χ² = 15.320, P < 0.0001; 50 mg/kg: 33.78 ± 2.60 ng/ml vs. 45.14 ± 4.10 ng/ml, χ² = 14.023, P < 0.0001, respectively), ALP (25 mg/kg: 2.35 ± 0.30 ng/ml vs. 3.28 ± 0.20 ng/ml, χ² = 4.101, P < 0.0001; 50 mg/kg: 2.34 ± 0.10 ng/ml vs. 3.28 ± 0.20 ng/ml, χ² = 2.033, P < 0.0001, respectively), and NO levels (25 mg/kg: 25.92% ± 2.30% vs. 38% ± 2.09%, χ² = 17.103, P < 0.0001; 50 mg/kg: 24.74% ± 4.10% vs. 38% ± 2.09%, χ² = 25.050, P = 0.001, respectively) compared to morphine group.

Conclusion: It seems that genistein administration might improve liver damages induced by morphine in mice.

Keywords: Genistein; Liver Damage; Morphine.

Figure 1

Forty-eight mice were equally divided into six groups. *Significant decrease of liver weight in morphine group compared to saline group (18.50 ± 0.90 g vs. 27.15 ± 0.50 g, χ² = 22.450, P = 0.001). ^†,‡Significant increase in genistein plus morphine groups compared to morphine group (25 mg/kg: 21.15 ± 2.13 g vs. 18.50 ± 0.90 g, χ² = 19.251, P < 0.0001; 50 mg/kg: 21.20 ± 1.00 g vs. 18.50 ± 0.90 g, χ² = 19.502, P < 0.0001, respectively).

Figure 2

Effects of genistein, morphine, and genistein plus morphine on the mean nitric oxide levels of 48 mice were equally divided into six groups. *Significant increase of nitric oxide in morphine group compared to saline group (38.00% ± 2.09% vs. 18.72% ± 4.40%, χ² = 20.404, P < 0.001). ^†,‡Significant decrease of genistein plus morphine-administrated groups compared to morphine group (25 mg/kg: 25.92% ± 2.30% vs. 38.00% ± 2.09%, χ² = 17.103, P < 0.0001; 50 mg/kg: 24.74% ± 4.10% vs. 38% ± 2.09%, χ² = 25.050, P = 0.001, respectively).

Figure 3

The morphometry of liver (a) Micrograph of the liver section in the saline group, (b) Micrograph of the liver section in morphine group, (c) Micrograph of the liver section in genistein (50 mg/kg) group, (d) Micrograph of liver section in morphine plus genistein (50 mg/kg) group (Haematoxylin Eosin, original magnification ×100). (e) *Significant increase of the hepatocytes and central vein diameters in morphine compared to saline group (diameter of hepatocytes: 3.03 ± 0.10 μm vs. 1.10 ± 0.05 μm, χ² = 9.873, P = 0.001 and central hepatic vein: 22.76 ± 1.90 μm vs. 15.04 ± 0.60 μm, χ² = 21.814, P = 0.001, respectively). ^†,‡Significant decrease in genistein plus morphine-administrated groups compared to morphine group (diameter of hepatocytes: 25 mg/kg: 2.17 ± 0.30 μm vs. 3.03 ± 0.10 μm, χ² = 22.780, P = 0.001; 50 mg/kg: 2.01 ± 0.20 μm vs. 3.03 ± 0.1 μm, χ² = 7.12, P = 0.001; central hepatic vein, 25 mg/kg: 19.53 ± 1.00 μm vs. 22.76 ± 1.90 μm, χ² = 20.681, P = 0.001; 50 mg/kg: 19.44 ± 1.20 μm vs. 22.76 ± 1.90 μm, χ² = 7.120, P = 0.001, respectively).