Showing papers on "Arecoline published in 2013"

Areca nut-induced buccal mucosa fibroblast contraction and its signaling: a potential role in oral submucous fibrosis—a precancer condition

Abstract: Betel quid (BQ) chewing is an oral habit that increases the risk of oral cancer and oral submucous fibrosis (OSF), a precancerous condition showing epithelial atrophy and tissue fibrosis. Persistent fibroblast contraction may induce the fibrotic contracture of tissue. In this study, we found that areca nut extract (ANE) (200-1200 µg/ml) stimulated buccal mucosa fibroblast (OMF)-populated collagen gel contraction. Arecoline but not arecaidine-two areca alkaloids, slightly induced the OMF contraction. Exogenous addition of carboxylesterase (2U/ml) prevented the arecoline- but not ANE-induced OMF contraction. OMF expressed inositol triphosphate (IP3) receptors. ANE-induced OMF (800 µg/ml) contraction was inhibited by U73122 [phospholipase C (PLC) inhibitor] and 2-aminoethoxydiphenyl borate (IP3 receptor antagonist), respectively. Ethylene glycol tetraacetic acid and verapamil, two calcium mobilization modulators, also suppressed the ANE-induced OMF contraction. ANE induced calcium/calmodulin kinase II and myosin light chain (MLC) phosphorylation in OMF. Moreover, W7 (a Ca(2+)/calmodulin inhibitor), HA1077 (Rho kinase inhibitor), ML-7 (MLC kinase inhibitor) and cytochalasin B (actin filament polymerization inhibitor) inhibited the ANE-induced OMF contraction. Although ANE elevated reactive oxygen species (ROS) level in OMF, catalase, superoxide dismutase and N-acetyl-L-cysteine showed no obvious effect on ANE-elicited OMF contraction. These results indicate that BQ chewing may affect the wound healing and fibrotic processes in OSF via inducing OMF contraction by ANE and areca alkaloids. AN components-induced OMF contraction was related to PLC/IP3/Ca(2+)/calmodulin and Rho signaling pathway as well as actin filament polymerization, but not solely due to ROS production.

Elevation of S100A4 Expression in Buccal Mucosal Fibroblasts by Arecoline: Involvement in the Pathogenesis of Oral Submucous Fibrosis

Abstract: Background S100A4, a member of the calcium-binding proteins, is dramatically elevated in a variety of fibrotic diseases. Areca quid chewing is the most important etiological factor in the pathogenesis of oral submucous fibrosis (OSF). OSF has been considered as a pre-cancerous condition of oral mucosa. The aim of this study was to determine the critical role of S100A4 expression in the pathogenesis of OSF both in vitro and in vivo.

Elevated Snail Expression Mediates Tumor Progression in Areca Quid Chewing-Associated Oral Squamous Cell Carcinoma via Reactive Oxygen Species

Abstract: Background Snail is an important transcription factor implicated in several tumor progression and can be induced by reactive oxygen species (ROS) Areca quid chewing is a major risk factor of oral squamous cell carcinoma (OSCC) Therefore, we hypothesize that the major areca nut alkaloid arecoline may induce Snail via ROS and involve in the pathogenesis of areca quid chewing-associated OSCC

Separation of polyphenols and arecoline from areca nut (Areca catechu L.) by solvent extraction, its antioxidant activity, and identification of polyphenols

Abstract: BACKGROUND Areca nut (Areca catechu L.) or betel nut, a commercial cash crop, is a rich source of polyphenols but also contains toxic alkaloids, mainly arecoline. Separation of these bioactive polyphenols from toxic constituents could propel the safe and beneficial use of betel nut; also it will help arecanut processing industries to produce arecoline-free products. With the aim to develop an effective method for maximum extraction of polyphenols with minimum arecoline, several factors such as nature of the solvent, pH (2–10), substrate concentration (6–14 %) and extraction time (30–150 min) under shaking conditions were evaluated. Qualitative analysis was done using spectrophotometry and high-performance liquid chromatography (HPLC). RESULTS Maximum extraction of polyphenols (407.47 mg GAE g−1), total tannin and its antioxidant activity with minimum arecoline (1.73 mg g−1 of sample) was achieved by using 80% acetone at pH 4 for 90 min with 10% w/v substrate under shaking conditions. CONCLUSION Solvent extraction under optimized parameters gave maximum polyphenols with minimum extraction of arecoline, and highest ratio of polyphenols to arecoline. HPLC and liquid chromatography–mass spectrometry results confirmed the presence of catechin and epicatechin in the extract, which suggests its potential as a source of bioactives. © 2013 Society of Chemical Industry

Toxicity of areca nut ingredients: activation of CHK1/CHK2, induction of cell cycle arrest, and regulation of MMP-9 and TIMPs production in SAS epithelial cells.

Abstract: Background There are 600 million betel quid chewers around the world. betel quid chewing is a major risk factor of oral cancer. Why betel quid components induce oral cancer is not clear. Methods Cytotoxicity of areca nut extract and arecoline (an areca nut alkaloid) to SAS oral epithelial cell line was evaluated by trypan blue dye exclusion and MTT assays. Cell cycle distribution and apoptosis was analyzed by propidium iodide staining flow cytometry. Chk1 and chk2 activation was analyzed by Pathscan phospho-enzyme-linked immunosorbent assay. Metalloproteinase-9 (MMP-9), tissue inhibitors of metalloproteinase (TIMPs) production was measured by enzyme-linked immunosorbent assay. Results Areca nut extract (800 μg/mL) and arecoline (>0.4 mmol/L) caused cell death, apoptosis, and cell cycle arrest of SAS cells. Areca nut extract and arecoline stimulated Chk1 and Chk2 phosphorylation in SAS cells. Areca nut extract stimulated cellular MMP-9 but suppressed TIMP-1 and TIMP-2 production. Conclusions Areca nut components activate Chk1/Chk2, alter cell cycle regulation/apoptosis, MMP-9, and TIMPs production, contributing to the pathogenesis of oral carcinogenesis. © 2012 Wiley Periodicals, Inc. Head Neck, 2013

Determination of arecoline in areca nut based on field amplification in capillary electrophoresis coupled with electrochemiluminescence detection

Abstract: A sensitive capillary electrophoresis-electrochemiluminescence (CE-ECL) assay with an ionic liquid (IL) was developed for the determination of arecoline in areca nut. The IL, 1-butyl-3-methylimidazolium tetrafluoroborate (BMImBF(4) ), was an effective additive improved not only the separation selectivity but also the detection sensitivity of the analyte. BMImBF(4) in the separation electrolyte made the resistance of the separation buffer much lower than that of the sample solution, which resulted in an enhanced field amplified electrokinetic injection CE. ECL intensity of arecoline is about two times higher than that of the analyte with phosphate-IL buffer system. Resolution between arecoline and other unknown compounds in real samples was improved. Under the optimized conditions (ECL detection at 1.2 V, 16 kV separation voltage, 20 mmol/L phosphate with 10 mmol/L BMImBF(4) buffer at pH 7.50, 5 mmol/L Ru(bpy)(3)(2+) and 50 mmol/L phosphate buffer in the detection reservoir), a detection limit of 5 × 10(-9) mol/L for arecoline was obtained. Relative standard deviations of the ECL intensity and the migration time were 4.51% and 0.72% for arecoline. This method was successfully applied to determination of the amount of arecoline in areca nut within 450 s.

Current Concepts about Areca Nut Chewing

Abstract: The habit of chewing areca is a habit of great antiquity. Theword ’areca’ is derived from the Malay word adakka (areca nut)or from adakeya, the Indian equivalent. Arecoline, the principalalkaloid in areca nut, acts as an agonist primarily at muscarinicacetylcholine receptors and stimulates the central and autonomicnervous system. This leads to subjective effects of increasedwell-being, alertness and stamina. It is known to improveconcentration and relaxation, with other reported effectsincluding lifting of mood, cariostatic property and also exerts adirect antimicrobial effect against bacteria. Arecaidine may haveanxiolytic properties through inhibition of gamma-amino butyricacid (GABA) reuptake.Despite these general effects, the adverse effects haveoutweighed them. Betal quid chewing is one of the major riskfactors of hepatocarcinoma, oropharyngeal and esophaguscancers. Arecoline, the main areca alkaloid of the betel nut, isreported to have cytotoxic, genotoxic and mutagenic effects invarious cells. It shows strong correlation to the incidence of oralsubmucosal fibrosis, leukoplakia and oral cancer, and has alsobeen found to impose toxic manifestations in immune, hepaticand other defense systems of the recipient.

The expression of O6‐methylguanine‐DNA methyltransferase in human oral keratinocytes stimulated with arecoline

Abstract: Background O6-methylguanine-DNA methyltransferase (MGMT) is a DNA repair enzyme that can protect cells from carcinogenic effects of alkylating agents by removing adducts from the O6 position of guanine. Evidences indicated that areca quid chewing may increase the risk of oral squamous cell carcinoma (OSCC). This study was to investigate the role of MGMT expression in OSCCs and the normal oral tissues. Methods Thirty-two OSCCs from areca quid chewers and ten normal oral tissue biopsy samples without areca quid chewing were analyzed by the immunohistochemistry for MGMT. Primary human oral keratinocytes (HOKs) were challenged with arecoline, the major alkaloid of areca nut, by Western blot. Nicotine, an important component of cigarette smoke, was added to find the possible regulatory mechanisms. Results Significant association was observed between low MGMT expression and advanced clinical stage of OSCCs and lymph node metastasis (P = 0.03). MGMT expression was significantly higher in patients only chewing areca quid than patients both chewing areca quid and smoking (P = 0.028). Arecoline was found to elevate MGMT expression in a dose- and time-dependent manner. The addition of nicotine was found to enhance arecoline-induced MGMT expression. Conclusion Our results indicate that MGMT could be used clinically as a predictive marker for tumor processing, the potential for lymph node metastasis as well as advanced clinical stage. MGMT expression was significantly upregulated by arecoline in HOKs. Nicotine has a synergistic effect of arecoline-induced MGMT expression. The cigarette smoking may act synergistically in the pathogenesis of OSCC in areca quid chewers via the upregulation of MGMT.

Transport of the areca nut alkaloid arecaidine by the human proton-coupled amino acid transporter 1 (hPAT1)

Abstract: Objectives The pyridine alkaloid arecaidine is an ingredient of areca nut preparations. It is responsible for many physiological effects observed during areca nut chewing. However, the mechanism underlying its oral bioavailability has not yet been studied. We investigated whether the H+-coupled amino acid transporter 1 (PAT1, SLC36A1), which is expressed in the intestinal epithelium, accepts arecaidine, arecoline, isoguvacine and other derivatives as substrates. Methods Inhibition of l-[3H]proline uptake by arecaidine and derivatives was determined in Caco-2 cells expressing hPAT1 constitutively and in HeLa cells transiently transfected with hPAT1-cDNA. Transmembrane transport of arecaidine and derivatives was measured electrophysiologically in Xenopus laevis oocytes. Key findings Arecaidine, guvacine and isoguvacine but not arecoline strongly inhibited the uptake of l-[3H]proline into Caco-2 cells. Kinetic analyses revealed the competitive manner of l-proline uptake inhibition by arecaidine. In HeLa cells transfected with hPAT1-cDNA an affinity constant of 3.8 mm was obtained for arecaidine. Electrophysiological measurements at hPAT1-expressing X. laevis oocytes demonstrated that arecaidine, guvacine and isoguvacine are transported by hPAT1 in an electrogenic manner. Conclusion We conclude that hPAT1 transports arecaidine, guvacine and isoguvacine across the apical membrane of enterocytes and that hPAT1 might be responsible for the intestinal absorption of these drug candidates.

Effect of arecoline on regeneration of injured peripheral nerves

Abstract: The present study provides in vitro and in vivo evaluation of arecoline on peripheral nerve regeneration. In the in vitro study, we found that arecoline at 50 μg/ml could significantly promote the survival and outgrowth of cultured Schwann cells as compared to the controls treated with culture medium only. In the in vivo study, we evaluated peripheral nerve regeneration across a 10-mm gap in the sciatic nerve of the rat, using a silicone rubber nerve chamber filled with the arecoline solution. In the control group, the chambers were filled with normal saline only. At the end of the fourth week, morphometric data revealed that the arecoline-treated group at 5 μg/ml significantly increased the number and the density of myelinated axons as compared to the controls. Immunohistochemical staining in the arecoline-treated animals at 5 μg/ml also showed their neural cells in the L4 and L5 dorsal root ganglia ipsilateral to the injury were strongly retrograde-labeled with fluorogold and lamina I-II regions in the dorsal horn ipsilateral to the injury were significantly calcitonin gene-related peptide-immunolabeled compared with the controls. In addition, we found that the number of macrophages recruited in the distal sciatic nerve was increased as the concentration of arecoline was increased. Electrophysiological measurements showed the arecoline-treated groups at 5 and 50 μg/ml had a relatively larger nerve conductive velocity of the evoked muscle action potentials compared to the controls. These results indicate that arecoline could stimulate local inflammatory conditions, improving the recovery of a severe peripheral nerve injury.

Arecoline suppresses HaCaT cell proliferation through cell cycle regulatory molecules

Abstract: Betel nut chewing is the most common cause of oral submucous fibrosis (OSF). Arecoline is the main component of the betel nut, and is associated with the occurrence and development of OSF through cytotoxicity, genotoxicity and DNA damage. Similar types of stimuli elicit differential responses in different cells. In the present study, we investigated the effects of arecoline on the HaCaT epithelial and Hel fibroblast cell lines. The data showed that arecoline affected HaCaT cell morphology. MTT assay revealed that arecoline suppressed HaCaT cell proliferation. Furthermore, we found that arecoline induced the cell cycle arrest of HaCaT cells. In comparison with the untreated control cells, following treatment with ≥75 µg/ml arecoline an increased percentage of HaCaT cells remained at the G0/G1 phase of the cell cycle, accompanied by a reduced percentage of cells in the S phase. However, arecoline treatment did not significantly alter Hel cell cycle distribution. In the HaCaT epithelial cells, arecoline downregulated expression of the G1/S phase regulatory proteins cyclin D1, CDK4, CDK2, E2F1 as determined by reverse transcription-PCR analysis and western blotting. In summary, arecoline inhibits HaCaT epithelial cell proliferation and survival, in a dose-dependent manner, and cell cycle arrest in the G1/S phase, while this is not obvious in the Hel fibroblast cells. Potentially, our findings may aid in the prevention of arecoline-associated human OSF.

Arecoline-stimulated placenta growth factor production in gingival epithelial cells: modulation by curcumin.

Abstract: Objective Placenta growth factor (PlGF) is associated with the progression and prognosis of oral cancer. Materials and Methods This study used ELISA, quantitative polymerase chain reaction, and Western blotting to study the arecoline-stimulated (PlGF) protein or mRNA expression in human gingival epithelial S-G cells. Results Arecoline, a major areca nut alkaloid and an oral carcinogen, could stimulate PlGF protein synthesis in S-G cells in a dose- and time-dependent manner. The levels of PlGF protein secretion increased about 3.1- and 3.8-fold after 24-h exposure to 0.4 and 0.8 mM arecoline, respectively. Pretreatment with antioxidant N-acetyl-l-cysteine (NAC) and ERK inhibitor PD98059, but not NF-κB inhibitor Bay 11-7082, JNK inhibitor SP600125, p38 MAPK inhibitor SB203580, and PI3-K inhibitor LY294002, significantly reduced arecoline-induced PlGF protein synthesis. ELISA analyses demonstrated that NAC and PD98059 reduced about 43% and 38% of the arecoline-induced PlGF protein secretion, respectively. However, combined treatment with NAC and PD98059 did not show additive effect. Moreover, 10 μM curcumin and 4 mM NAC significantly inhibited arecoline-induced ERK activation. Furthermore, 10 μM curcumin completely blocked arecoline-induced PlGF mRNA expression. Conclusion Arecoline-induced PlGF synthesis is probably mediated by reactive oxygen species/ERK pathways, and curcumin may be an useful agent in controlling oral carcinogenesis.

Arecoline inhibits interleukin-2 secretion in Jurkat cells by decreasing the expression of alpha7-nicotinic acetylcholine receptors and prostaglandin E2

Abstract: The purpose of the present study was to explore the effect of arecoline on phytohemagglutinin (PHA)-stimulated interleukin-2 (IL-2) secretion, the expression of alpha7-nicotinic acetylcholine receptors (α7-nAChRs), prostaglandin E2(PGE2) protein, and IL-2 mRNA in human lymphocyte cells (Jurkat cell line). The IL-2 and PGE2 were determined by enzyme-linked immunosorbent assay (ELISA). The expressions of phosphorylated extracellular signal-regulated kinase (ERK) and α7-nAChRs were determined by Western blotting. The level of IL-2 mRNA was determined by reverse-transcriptase polymerase chain reaction (RT-PCR). Arecoline, in a dose-dependent manner, significantly decreased IL-2 and PGE2 secretion by Jurkat cells incubated with 0 or 5 μg/ml 5 μg/ml PHA. PGE2 also significantly inhibited IL-2 secretion by Jurkat cells in a dose-dependent manner. In addition, reduced expression of PHA-induced ERK phosphorylation was observed in Jurkat cells treated with arecoline. PHA-enhanced IL-2 mRNA expression was also inhibited by arecoline. These results imply that arecoline inhibits the release of PGE2 and PHA-induced IL-2 secretion by Jurkat cells and that these effects seem to occur, at least in part, either through the attenuation of ERK in conjunction with a decrease of PHA-induced IL-2 mRNA expression. These results imply that arecoline inhibits the protein expression of α7-nAChRs , the release of PGE2 and PHA-induced IL-2 secretion by Jurkat cells.

A preliminary study of arecoline and guvacoline presence in the saliva of a "betel-quid" chewer using liquid-chromatography ion trap mass spectrometry.

Abstract: It has been reported that the parasympathomimetic alkaloid arecoline and the nootropic agent guvacoline have been detected in areca nut (Areca catechu L.) during extraction using a basic medium. Here, we have studied the detection of arecoline and guvacoline in vivo in saliva of a "betel-quid" chewer using liquid-chromatography ion trap mass spectrometry. In this paper, we provide evidence that guvacoline is absent in the neutral aqueous extract of betel nut, but is present in abundance in the aqueous extract with added time (pH 11.9). In an in vivo experiment, we demonstrated that guvacoline is present in the salivary extracts in the mouth with time (pH 9.5) and without lime (pH5.3).

Regulation of protease-activated receptor-1 expression in human buccal fibroblasts stimulated with arecoline.

Abstract: Background The purpose of this study was to compare the major thrombin receptor protease-activated receptor-1 (PAR-1) expression in normal human buccal mucosa and oral submucous fibrosis (OSF) specimens and further explore the potential mechanisms that may lead to induce PAR-1 expression. Methods Thirty OSF and 10 normal buccal mucosa specimens were examined by immunohistochemistry. Buccal mucosal fibroblasts (BMFs) were challenged with arecoline by using Western blot analysis. N-acetyl-L-cysteine (NAC), LY294002, herbimycin A, NS-398, and PD98059 were added to find the possible regulatory mechanisms. Results PAR-1 expression was significantly higher in OSF specimens (p < .05). Arecoline was found to elevate PAR-1 expression in a dose-dependent and time-dependent manner (p < .05). The addition of NAC, LY294002, herbimycin A, NS398, and PD98059 markedly inhibited the arecoline-induced PAR-1 expression (p < .05). Conclusion PAR-1 expression is significantly upregulated in areca quid chewing-associated OSF. Arecoline-induced PAR-1 expression was downregulated by NAC, LY294002, herbimycin A, NS398, and PD98059. © 2012 Wiley Periodicals, Inc. Head Neck, 2013

Research on effects of arecoline on refreshing and acute toxicity test

Abstract: Objective To study the betel nut actives arecoline refreshing and toxic effects,to provide reference basis for eating safety.Methods KM mice were randomly divided into three groups,intraperitoneal injection of various concentrations of arecoline solution 30 s after injection of phenobarbital sodium,the mouse anti-sleep time,while sterile saline and the same concentration of geranium control was recorded.The acute toxicity test used Horn method arecoline,to calculate its LD_(50) and to determine its toxicity level;Anatomy poisoned mice to observed the pathological changes.Results The anti-sleep time of arecoline was increasing with growth of concentration,0.5 mg of arecoline anti-1.5×10~(-4) g phenobarbital sodium-induced sleep time up to 38 min;The Arecoline the LD_(50) was 430 mg/kg·BW,confidence limits of 295-626 mg/kg·BW the middle poison.The poisoning death of the mice a slight enlargement of the liver and spleen become darker in color,larger size,lung color slightly darker,bleeding,intestinal vacuolation.Conclusion Arecoline the significant role of the anti-phenobarbital sodium hypnosis is a moderately toxic material.

MALDI-TOF mass spectrometry analysis of small molecular weight compounds (under 10 KDa) as biomarkers of rat hearts undergoing arecoline challenge.

Abstract: Context: Statistical and clinical reports indicate that betel nut chewing is strongly associated with progression of oral cancer because some ingredients in betel nuts are potential cancer promoters, especially arecoline. Early diagnosis for cancer biomarkers is the best strategy for prevention of cancer progression. Several methods are suggested for investigating cancer biomarkers. Among these methods, gel-based proteomics approach is the most powerful and recommended tool for investigating biomarkers due to its high-throughput. However, this proteomics approach is not suitable for screening biomarkers with molecular weight under 10 KDa because of the characteristics of gel electrophoresis.Objective: This study investigated biomarkers with molecular weight under 10 KDa in rats with arecoline challenge.Materials and methods: The centrifuging vials with membrane (10 KDa molecular weight cut-off) played a crucial role in this study. After centrifuging, the filtrate (containing compounds with molecular weigh...

A Simple and Rapid HPLC Technique for Determination of Arecoline in Areca Nut (Areca catechu L.) Extract

Abstract: A simple and rapid high-performance liquid chromatographic (HPLC) method for determination of arecoline extract from areca nut is reported. Arecoline is a major component cholinomimetic alkaloid found in areca nuts. It was extracted from dried seed powder to obtain high purity before analysis. The contents of arecoline in unripe and ripe areca nuts were compared. The analytical conditions for reverse-phase HPLC with UV detection were as follows: column, a 250 × 4.6 mm I.D., particle size 5 mm, Inertsil ® ODS-3 (GL sciences Inc., Tokyo, Japan); mobile phase, a 88:12 (%v/v) mixture of acetonitrile: phosphate buffer (pH 5.9); column temperature, 25 °C; flow rate, 1 mL/min; and detection at 254 nm. The retention time of arecoline was 5.0 min. The calibration curve of the method was linear (r 2 > 0.99) in the 10 - 200 mg/mL range. Method inter-day precision (RSD) was 0.42 - 1.15 %, and % recovery was 103.23 ± 5.76 % indicating high precision and accuracy of the method. The contents of arecoline in unripe and ripe areca nuts were 0.1434 ± 0.0016 and 0.0944 ± 0.0002 %w/w of dried seed powder, respectively. The method was simple, rapid, precise, accurate and selective to determine arecoline in areca nut extract.

Advance in studies on areca nuts and their active substances

Abstract: A number of clinical practices and studies indicated that areca nuts showed such effects as anthelmintic, food retention removal, qi activation and diuresis, and elimination of wetness and jaundice Arecoline is the most important pharmacological active ingredient for healthcare from areca plants with a wide influence on human functions In recent years, a lot of studies have been made on areca nuts and arecoline's pharmacology, physiology and immunity The article summarizes areca nuts and their active substances