Showing papers on "Arecoline published in 2015"

Nicotinic Activity of Arecoline, the Psychoactive Element of "Betel Nuts", Suggests a Basis for Habitual Use and Anti-Inflammatory Activity

Abstract: Habitual chewing of "betel nut" preparations constitutes the fourth most common human self-administration of a psychoactive substance after alcohol, caffeine, and nicotine. The primary active ingredient in these preparations is arecoline, which comes from the areca nut, the key component of all such preparations. Arecoline is known to be a relatively non-selective muscarinic partial agonist, accounting for many of the overt peripheral and central nervous system effects, but not likely to account for the addictive properties of the drug. We report that arecoline has activity on select nicotinic acetylcholine receptor (nAChR) subtypes, including the two classes of nAChR most related to the addictive properties of nicotine: receptors containing α4 and β2 subunits and those which also contain α6 and β3 subunits. Arecoline is a partial agonist with about 6-10% efficacy for the α4* and α6* receptors expressed in Xenopus oocytes. Additionally, arecoline is a silent agonist of α7 nAChR; while it does not activate α7 receptors when applied alone, it produces substantial activation when co-applied with the positive allosteric modulator PNU-120696. Some α7 silent agonists are effective inhibitors of inflammation, which might account for anti-inflammatory effects of arecoline. Arecoline's activity on nAChR associated with addiction may account for the habitual use of areca nut preparations in spite of the well-documented risk to personal health associated with oral diseases and cancer. The common link between betel and tobacco suggests that partial agonist therapies with cytisine or the related compound varenicline may also be used to aid betel cessation attempts.

Epithelial atrophy in oral submucous fibrosis is mediated by copper (II) and arecoline of areca nut

Abstract: Exposure of oral cavity to areca nut is associated with several pathological conditions including oral submucous fibrosis (OSF). Histopathologically OSF is characterized by epithelial atrophy, chronic inflammation, juxtaepithelial hyalinization, leading to fibrosis of submucosal tissue and affects 0.5% of the population in the Indian subcontinent. As the molecular mechanisms leading to atrophied epithelium and fibrosis are poorly understood, we studied areca nut actions on human keratinocyte and gingival fibroblast cells. Areca nut water extract (ANW) was cytotoxic to epithelial cells and had a pro-proliferative effect on fibroblasts. This opposite effect of ANW on epithelial and fibroblast cells was intriguing but reflects the OSF histopathology such as epithelial atrophy and proliferation of fibroblasts. We demonstrate that the pro-proliferative effects of ANW on fibroblasts are dependent on insulin-like growth factor signalling while the cytotoxic effects on keratinocytes are dependent on the generation of reactive oxygen species. Treatment of keratinocytes with arecoline which is a component of ANW along with copper resulted in enhanced cytotoxicity which becomes comparable to IC50 of ANW. Furthermore, studies using cyclic voltammetry, mass spectrometry and plasmid cleavage assay suggested that the presence of arecoline increases oxidation reduction potential of copper leading to enhanced cleavage of DNA which could generate an apoptotic response. Terminal deoxynucleotidyl transferase dUTP Nick End Labeling assay and Ki-67 index of OSF tissue sections suggested epithelial apoptosis, which could be responsible for the atrophy of OSF epithelium.

miR-203 inhibits arecoline-induced epithelial-mesenchymal transition by regulating secreted frizzled-related protein 4 and transmembrane-4 L six family member 1 in oral submucous fibrosis.

Abstract: Oral submucous fibrosis (OSF) is a potentially malignant disease predominantly found in Asian people. The areca nut has been implicated in this disease. Arecoline, one of the areca alkaloids, induces epithelial-mesenchymal transition (EMT)-related factors in primary human buccal mucosal fibroblasts. Yet, the mechanisms of the underlying arecoline-induced EMT in OSF remain unknown. In the present study, we aimed to investigate the role of microRNAs (miRNAs) in arecoline-induced EMT in HaCaT cells. We found that miR-203 was significantly downregulated in OSF tissues compared to that in normal buccal mucosa tissues, and that miR-203 negatively regulated secreted frizzled-related protein 4 (SFRP4) and positively regulated transmembrane-4 L six family member 1 (TM4SF1). We observed that upregulation of miR-203 significantly decreased the cell proliferation of HaCaT cells, and significantly upregulated the expression of cytokeratin 19 (CK19) and E-cadherin proteins, whereas it significantly downregulated the expression of N-cadherin and vimentin compared to these levels in the vehicle control cells. Thus, we provide evidence to illustrate that miR-203 plays a role in the pathogenesis of OSF, which may be a target for OSF management.

In silico Assessment of Drug-like Properties of Alkaloids from Areca catechu L Nut

Abstract: Purpose : To investigate in silico the drug-like properties of alkaloids (arecoline, arecaidine, guvacine, guvacoline, isoguvacine, arecolidine and homoarecoline) obtained from the fruits of Areca catechu L (areca nut). Methods : All chemical structures were re-drawn using Chemdraw Ultra 11.0. Furthermore, software including Bio-Loom for Windows - version 1.5, Molinspiration Property Calculator and ACD/I-LAB service were used to predict the drug-like properties of the alkaloids, including relative molecular mass (MW), partition coefficient log P (cLog P), number of hydrogen bond donors (HBD), number of hydrogen bond acceptors (HBA), topological polar surface area (TPSA), number of rotatable bonds (NROTB), pKa, and aqueous solubility at a given pH (LogS). In addition, Lipinski’s rule was used to evaluate druglike properties. Results : From our research, MWs of the seven compounds were all 7.5; furthermore, they possess good solubility at pH 1.0, 5.0, and 7.0. Conclusion : All the seven alkaloids possess good drug-like properties, and demonstrated good oral absorption and bioavailability. The results also suggest that these compounds can be further developed into new oral drugs for treating certain diseases. Keywords : Areca catechu L, Areca nut, Drug-like properties, Alkaloids, Arecoline, Arecaidine, Guvacine, Guvacoline, Isoguvacine, Arecolidine, Homoarecoline, In silico

Hypoxic regulation of plasminogen activator inhibitor-1 expression in human buccal mucosa fibroblasts stimulated with arecoline.

Abstract: Background Oral submucous fibrosis (OSF) is regarded as a pre-cancerous condition with fibrosis in oral subepithelial connective tissue. Hypoxia-inducible factor (HIF)-1α regulates a wide variety of profibrogenic genes, which are closely associated with tissue fibrosis. The aim of this study was to compare HIF-1α expression in normal buccal mucosa tissues and OSF specimens and further explore the potential mechanisms that may lead to the induction of HIF-1α expression. Method Twenty-five OSF specimens and six normal buccal mucosa were examined by immunohistochemistry. The expression of HIF-1α from fibroblasts cultured from OSF and normal buccal mucosa was measured by Western blot. Arecoline, a major areca nut alkaloid, was challenged to normal buccal mucosa fibroblasts (BMFs) to elucidate whether HIF-1α expression could affect by arecoline. In addition, the effects of arecoline on plasminogen activator inhibitor (PAI)-1 expression were evaluated in environmental hypoxia. Results HIF-1α expression was significantly higher in OSF specimens and expressed mainly by fibroblasts, epithelial cells, and inflammatory cells. Fibroblasts derived from OSF were found to exhibit higher HIF-1α protein expression than BMFs (P < 0.05). Arecoline was found to upregulate HIF-1α protein in a dose-dependent manner (P < 0.05). Hypoxia increased arecoline-induced PAI-1 protein expression than normoxic conditions (P < 0.05). Conclusion These results suggest that HIF-1α expression is significantly upregulated in OSF tissues from areca quid chewers, implying a potential role as a biomarker for local tissue hypoxia. The activation of HIF-1α may promote fibrogenesis by an increase of PAI-1 expression and a subsequent elevation of extracellular matrix production in oral submucosa leading to fibrosis.

Short-term exposure of zebrafish embryos to arecoline leads to retarded growth, motor impairment, and somite muscle fiber changes.

Abstract: The areca nut-chewing habit is common in Southeast Asia, India, and Taiwan, and arecoline is the most abundant and potent component in the areca nut. The effects of arecoline on birth defects have been explored in many species, including chicken, mice, and zebrafish. The effects of arecoline on embryos after long-term exposure are well established; however, the effects of short-term embryo exposure to arecoline are not understood. Using zebrafish, we study the effects of short-term exposure of arecoline on embryos to model the human habit of areca nut-chewing during early pregnancy. Arecoline, at concentrations from 0.001% to 0.04%, was administered to zebrafish embryos from 4 to 24 hours post fertilization. The morphological changes, survival rates, body length, and skeletal muscle fiber structure were then investigated by immunohistochemistry, confocal microscopy, and conventional electron microscopy. With exposure of embryos to increasing arecoline concentrations, we observed a significant decline in the hatching and survival rates, general growth retardation, lower locomotor activity, and swimming ability impairment. Immunofluorescent staining demonstrated a loose arrangement of myosin heavy chains, and ultrastructural observations revealed altered myofibril arrangement and swelling of the mitochondria. In addition, the results of flow-cytometry and JC-1 staining to assay mitochondria activity, as well as reverse transcription-polymerase chain reaction analyses of functional gene expression, revealed mitochondrial dysfunctions after exposure to arecoline. We confirmed that short-term arecoline exposure resulted in retarded embryonic development and decreased locomotor activity due to defective somitic skeletal muscle development and mitochondrial dysfunction.

Fibrotic Effects of Arecoline N-Oxide in Oral Potentially Malignant Disorders

Abstract: The metabolites of environmental chemicals play key roles in carcinogenesis. Areca nut is strongly associated with the development of oral potentially malignant disorders (OPMD) or cancer. The main alkaloid in the areca nut is arecoline, which is highly cytotoxic and genotoxic. Arecoline N-oxide, a metabolite of areca nut alkaloids, which has been identified in animal urine, has been shown to induce mutagenicity in bacteria. In this study, it was found that its protein adduct could be detected in oral keratinocytes treated with areca nut extract. Increased collagen expression and severity of squamous hyperplasia were observed in arecoline N-oxide treated mice. In cultured oral fibroblasts, arecoline N-oxide showed stronger effects on the increase of fibrotic related genes including TGF-beta1, S100A4, MMP-9, IL-6, and fibronectin and a decrease of E-cadherin as compared with arecoline. Finally, arecoline N-oxide stimulation effectively increased the DNA damage marker, gamma-H2A.X, both in vitro and in vivo. Taken together, these results indicate that arecoline N-oxide shows a high potential for the induction of OPMD.

Quantification of Salivary Arecoline, Arecaidine and N-Methylnipecotic Acid Levels in Volunteers by Liquid Chromatography-Tandem Mass Spectrometry

Abstract: Relatively little is known about the metabolism of areca nut in human saliva. We here describe the simultaneous quantification of areca nut metabolites: arecoline, arecaidine and N-methylnipecotic acid in saliva samples after chewing one 5 g areca nut by using liquid chromatography-tandem mass spectrometry (LC-MS-MS). Time courses of salivary areca nut metabolites in five adult male areca nut chewer volunteers were investigated. The limits of quantification were all 1.25 ng/mL for arecoline, arecaidine and N-methylnipecotic acid. Intra- and interday imprecisions were <4.2 and 13.6%, respectively. The within-day accuracy ranged from 82.2 to 116.7%, and the between-day accuracy ranged from 78.3 to 115.6%. Through areca nut chewing time course study, we found that salivary arecoline, arecaidine and N-methylnipecotic acid concentrations varied greatly over time between experiment individuals. Our findings suggest that arecoline might be metabolized slightly to arecaidine at 30 min after areca nut chewing and arecoline might be metabolized slightly to N-methylnipecotic acid at 25 min after areca nut chewing in the mouth. We first provide simultaneous quantification of human salivary arecoline, arecaidine and N-methylnipecotic acid levels using LC-MS-MS. This method may facilitate future research design in the pathogenic effects of areca nut exposure.

Arecoline stimulated early growth response-1 production in human buccal fibroblasts: Suppression by epigallocatechin-3-gallate

Abstract: Background Early growth response-1 (Egr-1) protein plays an important role in many human fibrotic diseases. Areca nut chewing is the most important risk factor of oral submucous fibrosis (OSF). Methods Egr-1 protein expression in OSF was examined using antibody to Egr-1. Arecoline-induced Egr-1 expression and its signaling pathways were assessed by Western blot analyses in human buccal mucosal fibroblasts (BMFs). Results Elevated Egr-1 staining was observed in epithelial cells, fibroblast, and inflammatory cells in 7 of 10 OSF cases. Arecoline, a main alkaloid found in the areca nut, stimulated Egr-1 synthesis in BMFs. Pretreatment with antioxidant N-acetyl-L-cysteine, c-Jun NH2-terminal kinase inhibitor SP600125, and extracellular signal-regulated kinase inhibitor PD98059 significantly reduced arecoline-induced Egr-1 synthesis. Epigallocatechin-3-gallate (EGCG) inhibited arecoline-induced Egr-1 synthesis and collagen gel contraction in a dose-responsive manner. Conclusion Constitutive Egr-1 expression during areca nut chewing may play a role in the pathogenesis of OSF. EGCG could be a good candidate for prevention or treatment of OSF. © 2014 Wiley Periodicals, Inc. Head Neck 37: 493–497, 2015

Arecoline increases basic fibroblast growth factor but reduces expression of IL‐1, IL‐6, G‐CSF and GM‐CSF in human umbilical vein endothelium

Abstract: Background Areca nut chewing is associated with oral submucous fibrosis (OSF). Raised vascular basic fibroblast growth factor may induce fibrosis. Arecoline is a muscarinic alkaloid in areca nut, which we earlier reported causes injury and necrosis of human endothelium. Materials and methods Human umbilical vein endothelial cells were exposed to arecoline with or without tumor necrosis factor-α, and separately to acetylcholine, muscarine, or nicotine. Protein levels of basic fibroblast growth factor, as well as the inflammatory cytokines: granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor, and Interleukins-6, 1-α and 1-β, were determined by enzyme-linked immunosorbent assay. mRNA levels were established by real-time reverse transcription polymerase chain reaction. Results Basic fibroblast growth factor was released into the culture medium at arecoline levels causing necrosis (P < 0.05). This contrasted with an opposite effect of arecoline on levels of the inflammatory cytokines (P < 0.05). Tumor necrosis factor-α increased IL-6 and granulocyte-macrophage colony stimulated factor, but arecoline reduced this stimulated expression (P < 0.05). Arecoline had no effect on mRNA for basic fibroblast growth factor, although there was reduced mRNA for the separate inflammatory cytokines studied. The effect of acetylcholine, muscarine, and nicotine was minimal and dissimilar to that of arecoline. Conclusions Data raise the possibility that arecoline-induced, vascular basic fibroblast growth factor contributes to OSF, by combining increased growth factor expression with endothelial necrosis, and thus driving fibroblast proliferation.

Elevated Lin28B expression is correlated with lymph node metastasis in oral squamous cell carcinomas

Abstract: Background Lymph node (LN) metastasis is the most common cause of oral squamous cell carcinoma (OSCC)-related death. Searching the detailed molecular mechanisms involved LN metastasis in OSCC is still an open question. Methods Paired tissue samples from tumor (T) and adjacent non-cancerous matched tissues (NCMT) parts, as well as LN metastatic lesions in patient with OSCC tissues were subjected to quantitative real-time PCR analysis for the expression levels of Lin28B. Arecoline, a major areca nut alkaloid, was to explore whether expression of Lin28B could be changed dose dependent in oral epithelial cells. Control and Lin28B-knockdown arecoline-stimulated oral epithelial cells were subjected to migration/invasion/anchorage-independent growth assay. Results Compared with NCMT samples from the same OSCC patient, the expression of Lin28B was increased in all of the tumor samples. A similar upregulation of Lin28B was also observed in LN metastatic when compared with local tumors. Arecoline treatment dose dependently induced Lin28B expression in SG and FaDu cells. Lentiviral-mediated silencing Lin28B expression significantly attenuated arecoline-induced oncogenicity including proliferation, migration, invasiveness, and anchorage-independent growth in SG and FaDu cells. Conclusions Lin28B may be a useful biomarker and novel molecular target for LN metastasis OSCC patients' treatment.

A protective role of arecoline hydrobromide in experimentally induced male diabetic rats.

Abstract: Objectives. Arecoline, the most potent and abundant alkaloid of betel nut, causes elevation of serum testosterone and androgen receptor expression in rat prostate, in addition to increase in serum insulin levels in rats, leading to insulin resistance and type 2 diabetes-like conditions. This study investigated the role of arecoline on the reproductive status of experimentally induced type 1 diabetic rats. Methods. Changes in the cellular architecture were analyzed by transmission electron microscopy. Blood glucose, serum insulin, testosterone, FSH, and LH were assayed. Fructose content of the coagulating gland and sialic acid content of the seminal vesicles were also analyzed. Results. Arecoline treatment for 10 days at a dose of 10 mg/kg of body weight markedly facilitated β-cell regeneration and reversed testicular and sex accessory dysfunctions by increasing the levels of serum insulin and gonadotropins in type 1 diabetic rats. Critical genes related to β-cell regeneration, such as pancreatic and duodenal homeobox 1 (pdx-1) and glucose transporter 2 (GLUT-2), were found to be activated by arecoline at the protein level. Conclusion. It can thus be suggested that arecoline is effective in ameliorating the detrimental effects caused by insulin deficiency on gonadal and male sex accessories in rats with type 1 diabetes.

Oral submucous fibrosis: a review article on etiopathogenesis.

Abstract: Areca quid chewing related oral mucosal lesions are potential hazard to a large population worldwide. Commercially freeze dried products such as pan masala, guthka and mawa have high concentration of areca nut per chew and appear to cause OSMF more rapidly than by self prepared conventional betel quid that contain smaller amounts of areca nut. The basic constituent of areca nut is either raw or dried or boiled or baked. Diverse agents including lime, tobacco, catechu, cloves, saffron and leaf of piper betel leaves may form a part of formulation. Many of the undesirable aspects of areca nut have been attributed to arecoline. These chemical appear to interfere with the molecular processes of deposition and or degradation of extracellular matrix molecules such as collagen, causing imbalance in the normal process. The most likely events that take place with regards to the above imbalance may be reduced phagocytosis of collagen by fibroblasts, up or down regulation of copper dependent enzyme lysyl oxidase, matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases . It has been postulated that areca nut may also induce the development of the disease by increased levels of cytokines in the lamina propria. Current evidence implicates collagen related genes in susceptibility and pathogenesis of OSMF. The individual mechanisms operating at various stages of the disease--initial, intermediate and advanced--need further study in order to propose appropriate therapeutic interventions.

Betel nut chewing associated with severe periodontitis

Abstract: Betel quid chewing has been in practice among people in many Asian countries since ages. It is the most widely used psychoactive substance and an important environmental risk factor for development of oral premalignant lesions and cancer. Arecoline, the major alkaloid of areca nut, has been known to cause cytotoxicity and genotoxicity in mammalian cells in vivo and in vitro and even contributes to carcinogenicity. It also increases the risk and severity of periodontal disease. This paper presents a case of betel nut chewing associated with severe periodontitis in a 32 year old male patient. DOI: http://dx.doi.org/10.3126/hren.v12i1.11990 Health Renaissance 2014;12(1):57-60

Preparation method of areca catechu leisure food

Abstract: The invention discloses a method for controlling cancerogens in areca catechu and preparing safe areca catechu leisure food. The method comprises the following steps: high-temperature and high-pressure cooking, ultrasonic pretreatment, marinating, baking, surface coloring, vacuum packaging, sterilization and the like. The method has the advantages that the content of arecoline and arecaidine which have the cancerigenic dangers in the areca catechu and the hardness of the products are greatly reduced, therefore, the variable hazard to lower fibers of an oral mucosa caused by areca catechu fibers is reduced, and the leisure food is safer in eating. The areca catechu leisure food prepared by the method has the advantages of low cost, good chewiness and mouthfeel, and stable property, and is well received by consumers.

Detection of arecoline by simple high-performance thin-layer chromatographic method in Indian nontobacco pan masala.

Abstract: Chewing the habit of blended pan masala containing areca nut with or without tobacco is a common practice in the Indian subcontinent. Arecoline, a pyridine alkaloid presence in areca nut alarmed for oral carcinogenesis and strictly prohibited in the western world. However, in India using blended pan masala is very popular among young and old individuals. In this context, we aimed to detect arecoline in Indian blended nontobacco pan masala sold in Kolkata using a simple densitometric high-performance thin-layer chromatographic (HPTLC) method and for alarming their use in common people. Eleven popularly Indian blended nontobacco pan masala were collected from the territory of Kolkata and isolated arecoline, following solvent extraction method derived for pyridine alkaloid. The quantitative analysis of arecoline was measured using automated software-based HPTLC instruments and validated the method according to International Conference on Harmonization guidelines. Arecoline was detected in all 11 blended nontobacco pan masala samples in a range of minimum 130 to maximum 415 μg/g dry samples. Arecoline is hazardous carcinogenic compound, so the use of Indian blended nontobacco pan masala should be restricted. Further, the method was found suitable for routine quantitative analysis of arecoline in areca nut containing substances.

Sensitization of nicotinic acetylcholine receptor of soil nematode caenorhabditis elegans through activation of muscarinic receptors by arecoline

Abstract: nematodes [6]. Nematodes were fed with E. coli OP50. All reagents used in this study were purchased from Sigma. Studies of the effects of levam-isole, nicotine, arecoline and atropine on the C. el-egans behavior were carried out at the temperature of 23°C using a stereoscopic microscope SMZ-05. Nematodes were incubated in 1 ml of NG buffer (0.3% NaCl, 1 mM CaCl 2 , 1 mM MgSO 4 , 25 mM K-phosphate buffer (pH 6.0)). The impairment of the swimming pattern induced by mechanical stimulation (shaking the test tube with a nematode) was considered a criterion of levamisole and nicotine effects on C. elegans behavior. These effects manifested themselves in the impairment of muscle coordination needed for sine-wave body movements, temporarily suspended swimming, swimming in a circle, etc. High concentrations of nicotine (0.7 mM), arecoline (3.5–7 mM) and at-ropine (2 mM) that were used in our experiments, like by some other authors [2–4, 7], can be justified by an extraordinarily low permeability of the nematode cuticle to many organic compounds as well as by a low rate of exchange between the in-The cholinergic system acquired by metazoans at the early stage of their evolution plays a key role in the regulation of basic functions of the body not only in vertebrates and humans but also in such primitive animals as Nematodes. In the soil nematode Caenorhabditis elegans, exactly like in mammals , the effect of acetylcholine (ACh) on target cells (neurons, locomotor muscles and other so-matic cells) is mediated by its binding to iono-tropic nicotinic receptors (N-cholinoreceptors) and metabotropic muscarinic receptors (M-cho-linoreceptors) [1–5]. A co-expression of N-and M-cholinoreceptor genes was found in the neu-rons of C. elegans [1, 2–4]. Hence, it was hypothesized that binding of ACh to M-cholinoreceptors can modulate its excitatory effect by changing N-cholinoreceptor sensitivity to ACh. The aim of this study was to verify this hypothesis. Experiments were conducted on young sexually mature nematodes of wild type N2 and the JD217 gar-3(vu78) mutant strain obtained from Caenorhabditis Genetics Center and bred in Petri dishes containing a standard growth medium for SHORT COMMUNICATIONS

Effects of arecoline on cell growth, migration, and differentiation in

Abstract: Background/purpose: Studies have supported a higher prevalence of periodontal disease among areca quid chewers than non-chewers. However, few studies have stated the effects of areca quid on periodontal tissues. The aim of this study was to investigate the inhib- itory effects of arecoline, the major alkaloid of areca nut, on murine immortalized cemento- blast cell line (OCCM.30).