Green coffee methanolic extract and silymarin protect against CCl4-induced hepatotoxicity in albino male rats

Abstract

Background: During the last few decades, patients worldwide have been interested in using alternative medicine in treating diseases to avoid the increased side effects of chemical medications. Green coffee is unroasted coffee seeds that have higher amounts of chlorogenic acid compared to roasted coffee. Green coffee was successfully used to protect against obesity, Alzheimer disease, high blood pressure and bacterial infection.

Methods: This study aimed to investigate the probable protective activity of the green coffee methanolic extract, silymarin and their combination on CCl₄-induced liver toxicity in male rats. Thirty Sprague - Dawley male albino rats were divided into 5 groups; control negative (G1) just got the vehicle (olive oil) and the other four groups received CCl₄ dissolved in olive oil through an intraperitoneal injection and were divided into untreated control positive group (G2), the third group (G3) was treated with green coffee methanolic extract, the fourth group (G4) was treated with silymarin, and the fifth group (G5) was treated with a combination of green coffee methanolic extract and silymarin.

Results: In the positive control group treated with CCl₄ (G2), the CCl₄-induced toxicity increased lipid peroxidation, IL-6, kidney function parameters, liver function enzymes, total cholesterol, triglycerides and low-density lipoproteins, and decreased irisin, antioxidants, CYP450 and high-density lipoprotein levels. Hepatic tissues were also injured. However, treating the injured rats in G3, G4 and G5 significantly improved the altered parameters and hepatic tissues.

Conclusions: Green coffee methanolic extract, silymarin, and their combination succeeded in protecting the male rats against CCl4 hepatotoxicity due to their antioxidant activity. Effect of green coffee methanolic extract mixed with silymarin in G5 was more efficient than that of green coffee methanolic extract in G3 or silymarin in G4.

Keywords: Antioxidants; Green coffee; Hepatotoxicity; Irisin; Oxidative stress.

Fig. 1

Effect of green coffee, silymarin and their combination on antioxidants and lipid peroxidation in CCl₄ induced hepatotoxic rats. a: catalase (CAT), b: glutathione transferase (GST),c: glutathione reduced (GSH), d:superoxide dismutase (SOD), e:total antioxidant capacity (TAC), f:malondialdehyde (MDA) G1: negative control group, G2: untreated positive control group, G3 treated with green coffee methanolic extract, G4 treated with silymarin and G5 treated with silymarin and green coffee methanolic extract

Fig. 2

Effect of green coffee methanolic extract, silymarin and their combination onIL-6, Irisin and P450 levels in CCl₄induced hepatotoxic male rats. a: Interleukin-6 (IL-6), b: Irisin,c:CYP450. G1: negative control group, G2: untreated positive control group, G3 treated with green coffee methanolic extract, G4 treated with silymarin and G5 treated with silymarin and green coffee methanolic extract

Fig. 3

a,b,c,d,e. Microscopic images of liver tissue in the groups under study. a: Control negative group, showing the normal histological structure of hepatic tissue with normal cells (red arrow) and vein (black arrow), b: Control positive group, showing fatty infiltrated and degenerated hepatocyte (red arrow), congested vein and sinusoids (black arrow), degenerated biliary duct (orange arrow) and bridging necrotic cells (blue arrow), c: Green coffee methanolic extract group, showing nearly normal hepatocytes (red arrow) with mildly inflammated vein (black arrow). d: Silymarin group, showing mild fatty degeneration, mild inflammation and interstitial edema (red arrow) with normal vein (black arrow). e: Combination of green coffee methanolic extract and silymarin group, showing nearly normal hepatocytes (red arrow) and vein (black arrow). (X 400, H&E stains)