Farnesol attenuates 1,2-dimethylhydrazine induced oxidative stress, inflammation and apoptotic responses in the colon of Wistar rats

https://doi.org/10.1016/j.cbi.2011.03.009Get rights and content

Abstract

Colon cancer is the major health hazard related with high mortality and it is a pathological consequence of persistent oxidative stress and inflammation. Farnesol, an isoprenoid alcohol, has been shown to possess antioxidant, anti-inflammatory and chemopreventive properties. The present study was performed to evaluate the protective efficacy of farnesol against 1,2-dimethylhydrazine (DMH) induced oxidative stress, inflammatory response and apoptotic tissue damage. Farnesol was administered once daily for seven consecutive days at the doses of 50 and 100 mg/kg body weight in corn oil. On day 7, a single injection of DMH was given subcutaneously in the groin at the dose of 40 mg/kg body weight. Protective effects of farnesol were assessed by using caspase-3 activity, tissue lipid peroxidation (LPO) and antioxidant status as end point markers. Further strengthening was evident on histopathological observations used to assess the protective efficacy of farnesol. Prophylactic treatment with farnesol significantly ameliorates DMH induced oxidative damage by diminishing the tissue LPO accompanied by increase in enzymatic viz., superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST) and quinone reductase (QR) and non-enzymatic viz., reduced glutathione (GSH) antioxidant status. Farnesol supplementation significantly decreased caspase-3 activity in colonic tissue. Histological findings also revealed that pretreatment with farnesol significantly reduced the severity of submucosal edema, regional destruction of the mucosal layer and intense infiltration of the inflammatory cells in mucosal and submucosal layers of the colon. The data of the present study suggest that farnesol effectively suppress DMH induced colonic mucosal damage by ameliorating oxidative stress, inflammatory and apoptotic responses.

Introduction

Colon cancer is the major health hazard related with high mortality and it is the second most common cause of cancer-related death [1]. Colon carcinogenesis is a multistep process and is thought to arise by the accretion of genetic alterations involving a variety of oncogenes and tumor suppressor genes that transform normal colonic epithelium into an invasive carcinoma [2]. Colon cancer is frequently a pathological consequence of persistent oxidative stress and inflammation [3], [4]. Oxidative stress is a state which occurs when the balance between the productions of reactive oxygen species (ROS) overcomes the endogenous antioxidant defense system and inflammation is a complex biological response of tissues to pathogens and damaged cells [5].

Several epidemiological studies suggest that diet is considered as one of the major factor associated with increased risk for colon cancer incidence and mortality [6], [7], [8]. Many experimental animal models have supported the idea that high fat diet augments the incidence of colon carcinogenesis [9], [10], [11] whereas low fat and high fiber (present in fruits and vegetables) diet, decreases the risk of colon cancer [12]. Many natural products present in the high fiber diets have been reported to possess chemopreventive properties against cancer [13]. Therefore, chemoprevention is a logical and current strategy to reduce the mortality from colon cancer because numerous chemopreventive agents are present in the diet [14].

Farnesol is a 15-carbon naturally occurring sesquiterpene and it may be endogenously generated in the cells by enzymatic dephosphorylation of farnesyl pyrophosphate, a metabolic precursor of squalene yielding sterols and other isoprenoid compounds [15], [16]. Dietary sources of farnesol are the plant products [17] including fruits and berries such as apricots, peaches, plums, blueberries, cranberries, raspberries and strawberries, vegetables such as tomatoes [18], herbs such as lemon grass and chamomile [19] and it is also obtained from the essential oils of ambrette seeds, and citronella [20]. Studies from our laboratory have revealed that farnesol is a potent antioxidant and protects the kidneys and lungs against oxidative damage induced by Ferric-Nitrilo Tri Acetate (Fe-NTA) and Cigarrette Smoke Extract (CSE), respectively [21], [22]. Previous studies also have been shown that farnesol exhibits significant anti-tumor and anti-carcinogenesis effects in vivo that might be due to the inhibition of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase [23], [24], [25], [26], [27]. Thus farnesol results in the inhibition of cholesterol formation due to HMG CoA reductase inhibition and may alter the cell/mitochondrial membrane integrity thereby leads to the apoptosis favorably in tumor cells as compared to normal cells [26], [28], [29], [30], [31]. Tumor cells are highly proliferating cells requiring high levels of cholesterol to maintain the cell/mitochondrial membrane integrity [25], [26]. Apoptosis, is one of the forms of programmed cell death, characterized by chromatin condensation, nuclear fragmentation, membrane blebbing, cytoskeletal rearrangement and cell shrinkage. It is involved in many physiological and pathological processes and helps to regulate tissue homeostasis by eliminating potentially deleterious cells [32]. Oxidative stress is known to be involved in the induction of apoptosis and antioxidants have been reported to diminish the extent of apoptotic tissue damages [33], [34].

1,2-Dimethylhydrazine (DMH) is a colon specific carcinogen and it has been widely used to induce colon cancer in rodents [35]. DMH undergoes metabolism mainly in the liver and to some extent also in the colon and the ultimate metabolite thus formed in the liver is delivered to the colon via, blood or bile, as glucoronide conjugates [36], [37]. In vivo transformation of DMH results in the formation of azomethane and N-oxidation of azomethane leads to the formation of azoxymethane. Further, hydroxylation of azoxymethane leads to the formation of methyazoxymethanol which is an unstable compound and readily yields highly reactive electrophilic methyldiazonium ion. The latter leads to the formation of methyl free radicals and DMH also generates hydroxyl radical or hydrogen peroxide in the presence of metal ion which are known to elicit oxidative stress due to imbalance between the production of ROS and endogenous antioxidants [38], [39]. It has been reported earlier that these ROS are mainly responsible for the damaging effects of the DMH in colonic tissue [38]. DMH also causes covalent modification of DNA by 8-hydroxy-2′-deoxyguanosine (8-OHdG) adduct formation which is a marker of oxidative DNA damage [40] and it has been well accepted that oxidative DNA damage plays an important role in carcinogenesis [41], [42]. DMH-induced colon tumorigenesis reflects many of the same cell kinetic and similar molecular and histopathological alterations to the sporadic colon tumors of humans [43], [44], [45], [46].

The present study was intended to explore the anticipatory effects of farnesol against DMH induced oxidative stress, inflammation and apoptotic responses in the colon of Wistar rats.

Section snippets

Chemicals

Reduced glutathione (GSH), oxidized glutathione (GSSG), nicotinamide adenine dinucleotide phosphate reduced (NADPH), flavin adenine dinucleotide (FAD), ethylene diamine tetra acetic acid (EDTA), nicotinamide adenine dinucleotide reduced (NADH), thiobarbituric acid (TBA), trichloroacetic acid (TCA), bovine serum albumin (BSA), 5,5’-dithiobis-(2-nitrobenzoic acid) (DTNB), 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dimethylhydrazine (DMH), glutathione reductase were obtained from Sigma (Sigma

Effect of prophylactic treatment of farnesol on the activities of glutathione dependent enzymes in colonic tissue

The activities of GPx, GR and GST decreased significantly (p < 0.001) in Group II as compared to Group I. Farnesol pretreatment at the dose of 50 mg/kg body weight significantly increased the activities of GPx (p < 0.01), GST (p < 0.001), and GR (p < 0.01) in Group III as compared to Group II. Higher dose of farnesol (100 mg/kg body weight) also showed significant increase in the activities of GPx (p < 0.001), GST (p < 0.001), and GR (p < 0.01) in Group IV as compared to Group II. However, the activities of

Discussion

A current upsurge in classifying natural products as cancer chemopreventive agents is gaining much attention of many investigators because natural products like fruits, vegetables, medicinal plants, and herbs have many pharmacological properties and have potential to fight against numerous human diseases associated with oxidative stress. Several natural compounds of dietary sources have been reported to inhibit various diseases including cancer [57], [58] and farnesol is one of them, belongs to

Conclusion

The precise mechanism of chemopreventive action of farnesol against colon carcinogenesis remains to be elucidated but the plausible mechanism of the protection of farnesol may be through the induction of the antioxidant enzymes. From the findings of the current study it can be concluded that farnesol supplementation effectively suppressed the initial phases of colon carcinogenesis probably by reducing the oxidative damage, inflammatory and apoptotic responses induced by DMH in rats. The results

Conflict of interest statement

None declared.

Acknowledgment

We are thankful to the University Grants Commission (UGC) Government of India for the financial support to the first author.

References (84)

  • T.E. Meigs et al.

    Regulation of 3-hydroxy- 3-methylglutaryl-CoA reductase degradation by the nonsterol mevalonate metabolite farnesol in vivo

    J. Biol. Chem.

    (1996)
  • C.V. Rao et al.

    Chemopreventive effect of farnesol and lanosterol on colon carcinogenesis

    Cancer Detect. Prev.

    (2002)
  • T.M. Buttke et al.

    Oxidative stress as a mediator of apoptosis

    Immunol. Today

    (1994)
  • A.F. Slater et al.

    The role of intracellular oxidants in apoptosis

    Biochim. Biophys. Acta

    (1995)
  • B. Halliwell et al.

    Role of free radicals and catalytic metal ions in human disease: an overview

    Methods Enzymol.

    (1990)
  • K.M. Pozharisski et al.

    Experimental intestinal cancer research with special reference to human pathology

    Adv. Cancer Res.

    (1979)
  • I. Carlberg et al.

    Glutathione level in rat brain

    J. Biol. Chem.

    (1975)
  • W.H. Habig et al.

    Glutathione-S-transferases: the first enzymatic step in mercapturic acid formation

    J. Biol. Chem.

    (1974)
  • J.R. Wright et al.

    Cytosolic factors which affect microsomal lipid peroxidation in lung and liver

    Arch. Biochem. Biophys.

    (1981)
  • O.H. Lowry et al.

    Protein measurement with the Folin phenol reagent

    J. Biol. Chem.

    (1951)
  • L.W. Wattenberg

    Chemoprevention of cancer

    Prev. Med.

    (1996)
  • B.C. Pence

    Dietary selenium and antioxidant status: toxic effects of 1,2-dimethylhydrazine in rats

    J. Nutr.

    (1991)
  • P.K. Dudeja et al.

    1,2-Dimethylhydrazine-induced alterations in lipid peroxidation in preneoplastic and neoplastic colonic tissues

    Biochim. Biophys. Acta

    (1990)
  • A. Seven et al.

    Evaluation of oxidative stress parameters in blood of patients with laryngeal carcinoma

    Clin. Biochem.

    (1999)
  • T.M. Yau

    Mutagenicity and cytotoxicity of malonaldehyde in mammalian cells

    Mech. Ageing Dev.

    (1979)
  • H.J. Forman et al.

    Glutathione: overview of its protective roles, measurement, and biosynthesis

    Mol. Aspects Med.

    (2009)
  • S. Sharma et al.

    Modulatory effect of soy isoflavones on biochemical alterations mediated by TPA in mouse skin model

    Food Chem. Toxicol.

    (2004)
  • N.V. Rajeshkumar et al.

    Modulation of carcinogenic response and antioxidant enzymes of rats administered with 1,2-dimethylhydrazine by picroliv

    Cancer Lett.

    (2003)
  • D. Dreher et al.

    Role of oxygen free radicals in cancer development

    Eur. J. Cancer

    (1996)
  • T.L. Horn et al.

    Modulation of hepatic and renal drug metabolizing enzyme activities in rats by subchronic administration of farnesol

    Chem. Biol. Interact.

    (2005)
  • J.H. Joo et al.

    Molecular mechanisms involved in farnesol-induced apoptosis

    Cancer Lett.

    (2010)
  • S.C. Chaudhary et al.

    Chemopreventive effect of farnesol on DMBA/TPA-induced skin tumorigenesis: involvement of inflammation, Ras-ERK pathway and apoptosis

    Life Sci.

    (2009)
  • J. Wen et al.

    Oxidative stress-mediated apoptosis: the anticancer effect of the sesquiterpene lactone parthenolide

    J. Biol. Chem.

    (2002)
  • S.C. Chaudhary et al.

    Chemopreventive effect of farnesol on DMBA/TPA-induced skin tumorigenesis: involvement of inflammation, Ras-ERK pathway and apoptosis

    Life Sci.

    (2009)
  • C.A. O’Brien et al.

    A human colon cancer cell capable of initiating tumour growth in immunodeficient mice

    Nature

    (2007)
  • P.A. Janne et al.

    Chemoprevention of colorectal cancer

    N. Engl. J. Med.

    (2000)
  • J. Terzic et al.

    Inflammation and colon cancer

    Gastroenterology

    (2010)
  • L. Ferrero-Miliani et al.

    Chronic inflammation: importance of NOD2 and NALP3 in interleukin-1beta generation

    Clin. Exp. Immunol.

    (2007)
  • R. Doll

    General epidemiologic considerations in etiology of colorectal cancer

  • M.J. Burstein

    Dietary factors related to colorectal neoplasms

    Surg. Clin. N. Am.

    (1993)
  • B.R. Bansal et al.

    Effects of diet on colon carcinogenesis and the immune system in rats treated with 1,2-dimethylhydrazine

    Cancer Res.

    (1978)
  • B.S. Reddy et al.

    Effect of a diet with high levels of protein and fat on colon carcinogenesis in F344 rats treated with 1,2-dimethylhydrazine

    J. Natl. Cancer Inst.

    (1976)
  • Cited by (124)

    View all citing articles on Scopus
    View full text