Ameliorative effects of Moringa oleifera Lam seed extract on liver fibrosis in rats

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Abstract

This study was carried out to evaluate the effect of Moringa oleifera Lam (Moringa) seed extract on liver fibrosis. Liver fibrosis was induced by the oral administration of 20% carbon tetrachloride (CCl4), twice weekly and for 8 weeks. Simultaneously, M. oleifera Lam seed extract (1 g/kg) was orally administered daily. The biochemical and histological results showed that Moringa reduced liver damage as well as symptoms of liver fibrosis. The administration of Moringa seed extract decreased the CCl4-induced elevation of serum aminotransferase activities and globulin level. The elevations of hepatic hydroxyproline content and myeloperoxidase activity were also reduced by Moringa treatment. Furthermore, the immunohistochemical study showed that Moringa markedly reduced the numbers of smooth muscle α-actin-positive cells and the accumulation of collagens I and III in liver. Moringa seed extract showed significant inhibitory effect on 1,1-diphenyl-2-picrylhydrazyl free radical, as well as strong reducing antioxidant power. The activity of superoxide dismutase as well as the content of both malondialdehyde and protein carbonyl, which are oxidative stress markers, were reversed after treatment with Moringa. Finally, these results suggested that Moringa seed extract can act against CCl4-induced liver injury and fibrosis in rats by a mechanism related to its antioxidant properties, anti-inflammatory effect and its ability to attenuate the hepatic stellate cells activation.

Introduction

Liver fibrosis is a dynamic wound-healing response to chronic hepatocellular damage that represents a major medical problem with significant morbidity and mortality. It is characterized by an excessive deposition of extracellular matrix (ECM) proteins of which types I and III collagens are predominant. This excess deposition of ECM disrupts the normal architecture of the liver that alters the normal function of the organ, resulting in pathophysiological damage to the organ (Tsukada et al., 2006, Torok, 2008). Hepatic stellate cells (HSCs) are presently regarded as one of the key cell types involved in the progression of liver fibrosis (Gutierrez-Ruiz and Gomez-Quiroz, 2007). Indeed, HSC activation to a proliferative, myofibroblastic phenotype plays a key role in hepatic fibrogenesis, since these cells are the principal cellular source of the excess collagen synthesis during hepatic fibrosis (Gressner et al., 2007, Luk et al., 2007). Cellular changes accompanying HSC activation include morphological changes such as the appearance of the cytoskeletal protein smooth muscle α-actin (α-SMA) and a dramatic increase in types I and III collagens.

Efforts have been made to search for effective anti-fibrotic agents. Medicinally, natural drugs have made a significant contribution to the treatment of liver fibrosis. Use of herbal drugs in the treatment of liver diseases has a long tradition, especially in Eastern medicine (Schuppan et al., 1999, Luk et al., 2007). Among myriad of natural plants, Moringa oleifera Lam, is one of the best known and most distributed species of Moringacae family. Moringa is an important tropical crop that is used as human food, medicine and in oil production. It is a small sized tree, which is native to south Asia, Africa and Arabia (Anwer et al., 2007). It is commonly known as drumstick tree or horse radish tree. While in the Nile valley, the name of tree is Shagara al Rauwaq, which means tree of purifying (Anwer et al., 2007). Medicinally, various parts of Moringa are generally known for their multiple pharmacological effects including their antitumor (Guevara et al., 1999, Bharali et al., 2003), antihyperglycemic (Anwer et al., 2007) and anti-inflammatory (Mahajan and Mehta, 2008) effects. Furthermore, the extract of Moringa has been shown to have potent antioxidant action in vivo (Kumar and Pari, 2003, Arabshahi et al., 2007) studies and the hepatoprotective effects of an ethanolic extract of Moringa seeds was investigated against diclofenac-induced acute liver damage in male rats (Hamza, 2007). Phytochemical investigations to isolate the bioactive compounds from the seeds of Moringa resulted in the isolation of glycosides such as 4(alpha-l-rhamnosyloxy-benzylglucosinolate), niazimicin and niazirin, beta-sitosterol and Moringa oil (Guevara et al., 1999, Lalas and Tsaknis, 2002, Bennet et al., 2003).

This investigation was designed to evaluate the protective effects of seeds from Moringa on hepatic fibrosis in male rats induced by CCl4 and its relationship with oxidative stress, inflammation, and HSC activation.

Section snippets

Chemicals

Chloramin T, type I and III collagens, N-methyl-2-phenylindol, Folin–Ciocalteu reagent, pyrogallol, superoxide dismutase, catalase, 2,4-dinitophenylhydrazine, o-dianisidine, p-dimethyl-amino-benzaldehyde and bovine albumin were obtained from Sigma Chemical Co. (St. Louis, MO, USA), and α-smooth muscle actin (Neomarkers, USA) and all other chemicals were obtained from local commercial suppliers.

Plant material

The seeds of Moringa were gathered in May 2008, from different localities in the city of Al-Ain,

Serum markers of liver damage

Table 1 presents the effects of Moringa and silymarin on CCl4-treated in rats. In the CCl4-intoxicated group, serum activities of AST and ALT were markedly increased (P < 0.001) and globulin levels were significantly increased (P < 0.05) while serum albumin levels were considerably decreased (P < 0.001) compared to that of the control group. In contrast, treatment with Moringa has completely blocked the CCl4-induced elevation of serum ALT and AST activities. Furthermore it is markedly abolished the

Discussion

Liver fibrosis is a dynamic and regulated wound-healing response to chronic hepatocellular damage that represents a major medical problem with significant morbidity and mortality (Tsukada et al., 2006, Gressner et al., 2007). This study showed that Moringa has a remarkable protective effect against CCl4-induced liver fibrosis in rats. This result was confirmed by the histological findings and biochemical analysis of HP, a marker of collagen deposition in liver. It is well known that hepatic

Conflict of interest statement

The authors declare that there are no conflicts of interest.

Acknowledgements

The author is thankful to the Research Committee, UAE University, for providing funds to carry out this work. The author also thanks Prof. Waleed Hamza, Head of the Biology Department, UAE University for his valuable help in facilitating this research in the department. The author is grateful to Mr. Sayel Daoud (Twam hospital, UAE) for his kind help with histological preparations. The author is thankful to Dr. Tom Fowler (UAE University, UAE) and Dr. Hosam Gharib Abdelhady (University of

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