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Ming-yung Chou - One of the best experts on this subject based on the ideXlab platform.
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Cytotoxic and non‐genotoxic effects of Arecoline on human buccal fibroblasts in vitro
Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pat, 2007Co-Authors: Yu-chao Chang, Kuo-wei Tai, Min-hsiung Cheng, Lin Shin-shen Chou, Ming-yung ChouAbstract:Chang YC, Tai KW, Cheng MH, Chou LSS, Chou MY: Cytotoxic and non-genotoxic effects of Arecoline on human buccal fibroblasts in vitro. J Oral Pathol Med 1998; 27: 68–71. © Munksgaard, 1998. Betel quid chewing has been linked to oral submucous fibrosis and oral cancer. Cytotoxicity and genotoxicity assays were used to investigate the pathobiologi-cal effects of Arecoline on cultured human buccal fibroblasts. Arecoline increased double-stranded polynucleic acid at the concentration of 0.1 to 10 μg/ml in a concentration-dependent manner. At a concentration higher than 50 μg/ml, Arecoline was cytotoxic to cultured fibroblasts and the cytotoxicity was dose-dependent. No genotoxicity for Arecoline was found even at a concentration of 400 μg/ml. On the other hand, 600 μg/ml glutathione (GSH) and 200 μg/ml glycyrrhizin could prevent the Arecoline-induced cytotoxicity. These results indicate that Arecoline is a cytotoxic agent and no genotoxicity was found to human buccal fibroblasts. Furthermore, increasing consumption of GSH- and glycyrrhizin-rich foods may reduce the oral diseases associated with betel quid chewing.
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cytotoxic and non genotoxic effects of Arecoline on human buccal fibroblasts in vitro
Journal of Oral Pathology & Medicine, 2007Co-Authors: Yu-chao Chang, Kuo-wei Tai, Min-hsiung Cheng, Lin Shin-shen Chou, Ming-yung ChouAbstract:Chang YC, Tai KW, Cheng MH, Chou LSS, Chou MY: Cytotoxic and non-genotoxic effects of Arecoline on human buccal fibroblasts in vitro. J Oral Pathol Med 1998; 27: 68–71. © Munksgaard, 1998. Betel quid chewing has been linked to oral submucous fibrosis and oral cancer. Cytotoxicity and genotoxicity assays were used to investigate the pathobiologi-cal effects of Arecoline on cultured human buccal fibroblasts. Arecoline increased double-stranded polynucleic acid at the concentration of 0.1 to 10 μg/ml in a concentration-dependent manner. At a concentration higher than 50 μg/ml, Arecoline was cytotoxic to cultured fibroblasts and the cytotoxicity was dose-dependent. No genotoxicity for Arecoline was found even at a concentration of 400 μg/ml. On the other hand, 600 μg/ml glutathione (GSH) and 200 μg/ml glycyrrhizin could prevent the Arecoline-induced cytotoxicity. These results indicate that Arecoline is a cytotoxic agent and no genotoxicity was found to human buccal fibroblasts. Furthermore, increasing consumption of GSH- and glycyrrhizin-rich foods may reduce the oral diseases associated with betel quid chewing.
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elevated vimentin expression in buccal mucosal fibroblasts by Arecoline in vitro as a possible pathogenesis for oral submucous fibrosis
Oral Oncology, 2002Co-Authors: Yu-chao Chang, Kuo-wei Tai, Ming-yung Chou, Chung-hung Tsai, S F Yang, Chong-kuei LiiAbstract:Areca quid chewing is strongly correlated with oral submucous fibrosis (OSF) in Taiwan. The cytotoxicity of Arecoline, a major areca nut alkaloid, on human oral fibroblasts has been extensively studied. To date, however, there has been little research exploring the possible effects of Arecoline on cytoskeleton components. In this study, in addition to conducting a cytotoxicity assay, we examine the effect of Arecoline on vimentin, an intermediate filament, and its expression in human buccal mucosal fibroblasts on exposure to various levels of Arecoline (0–200 μg/ml) for 48 h. At a concentration above 50 μg/ml, Arecoline demonstrated dose-dependent cytotoxicity (P<0.05) for cultured fibroblasts. Using sodium dodecyl sulphate-polyacrylamide gel electrophoresis, we demonstrated dose-dependent elevation of 57 kDa cytoskeletal-protein levels for Arecoline. Evidence from immunoblotting assay indicated this 57 kDa cytoskeletal protein was vimentin. The increase in vimentin with Arecoline exposure corresponded to that noted for fibroblasts cultured from OSF patients. Immunohistochemical assay also revealed that vimentin expression was much higher for OSF specimens than for normal buccal mucosa. We suggest these results may advance understanding of the possible pathogenesis for submucous fibrosis through the transformation of normal buccal mucosa as a result of areca quid chewing.
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Adverse effects of Arecoline and nicotine on human periodontal ligament fibroblasts in vitro
Journal of clinical periodontology, 2001Co-Authors: Yu-chao Chang, Chong-kuei Lii, Kuo-wei Tai, Ming-yung ChouAbstract:BACKGROUND, AIMS The habit of betel nut chewing impinges on the daily lives of approximately 200 million people. Betel quid chewers have a higher prevalence of periodontal diseases than non-chewers. This study examined the pathobiological effects of Arecoline, a major component of the betel nut alkaloids, on human periodontal ligament fibroblasts (PDLF) in vitro. METHOD Cell viability, proliferation, protein synthesis, and cellular thiol levels were used to investigate the effects of human PDLF exposed to Arecoline levels of 0 to 200 microg/ml. In addition, nicotine was added to test how it modulated the effects of Arecoline. RESULTS Arecoline significantly inhibited cell proliferation in a dose-dependent manner. At concentrations of 10 and 30 microg/ml, Arecoline suppressed the growth of PDLF by 20% and 50% (p < 0.05), respectively. Arecoline also decreased protein synthesis in a dose-dependent manner during a 24-h culture period. A 100 microg/ ml concentration level of Arecoline significantly inhibited protein synthesis to only 50% of that in the untreated control (p < 0.05). Moreover, Arecoline significantly depleted intracellular thiols in a dose-dependent manner. At concentrations of 25 microg/ml and 100 microg/ml, Arecoline depleted about 18% and 56% of thiols (p < 0.05), respectively. This suggests that Arecoline itself might augment the destruction of periodontium associated with betel nut use. Furthermore, the addition of nicotine acted with a synergistic effect on the Arecoline-induced cytotoxicity. At a concentration of 60 microg/ml, Arecoline suppressed the growth of PDLF by about 33% and 5 mM nicotine enhanced the Arecoline-induced cytotoxic response to cause about 66% cell death. CONCLUSION During thiol depletion, Arecoline may render human PDLF more vulnerable to reactive agents within cigarettes. Taken together, people who combine habits of betel nut chewing with cigarette smoking could be more susceptible to periodontium damage than betel nut chewing alone.
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synergistic effects of nicotine on Arecoline induced cytotoxicity in human buccal mucosal fibroblasts
Journal of Oral Pathology & Medicine, 2001Co-Authors: Yu-chao Chang, Chong-kuei Lii, Kuo-wei Tai, Tsuihwa Tseng, Ming-yung ChouAbstract:Areca quid chewing has been linked to oral submucous fibrosis and oral cancer. Arecoline, a major areca nut alkaloid, is considered to be the most important etiologic factor in the areca nut. In order to elucidate the pathobiological effects of Arecoline, cytotoxicity assays, cellular glutathione S-transferase (GST) activity and lipid peroxidation assay were employed to investigate cultured human buccal mucosal fibroblasts. To date, there is a large proportion of areca quid chewers who are also smokers. Furthermore, nicotine, the major product of cigarette smoking, was added to test how it modulated the cytotoxicity of Arecoline. At a concentration higher than 50 microg/ml, Arecoline was shown to be cytotoxic to human buccal fibroblasts in a dose-dependent manner by the alamar blue dye colorimetric assay (P<0.05). In addition, Arecoline significantly decreased GST activity in a dose-dependent manner (P<0.05). At concentrations of 100 microg/ml and 400 microg/ml, Arecoline reduced GST activity about 21% and 46%, respectively, during a 24 h incubation period. However, Arecoline at any test dose did not increase lipid peroxidation in the present human buccal fibroblast test system. The addition of extracellular nicotine acted synergistically on the Arecoline-induced cytotoxicity. Arecoline at a concentration of 50 microg/ml caused about 30% of cell death over the 24 h incubation period. However, 2.5 mM nicotine enhanced the cytotoxic response and caused about 50% of cell death on 50 microg/ml Arecoline-induced cytotoxicity. Taken together, Arecoline may render human buccal mucosal fibroblasts more vulnerable to other reactive agents in cigarettes via GST reduction. The compounds of tobacco products may act synergistically in the pathogenesis of oral mucosal lesions in areca quid chewers. The data presented here may partly explain why patients who combined the habits of areca quid chewing and cigarette smoking are at greater risk of contracting oral cancer.
Lea-yea Chuang - One of the best experts on this subject based on the ideXlab platform.
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Arecoline induced phosphorylated p53 and p21 waf1 protein expression is dependent on atm atr and phosphatidylinositol 3 kinase in clone 9 cells
Journal of Cellular Biochemistry, 2009Co-Authors: Wen-wen Chou, Jinn-yuh Guh, Jung-fa Tsai, Chi-ching Hwang, Sheanjaw Chiou, Lea-yea ChuangAbstract:Betel-quid use is associated with liver cancer whereas its constituent Arecoline is cytotoxic, genotoxic, and induces p53-dependent p21(WAF1) protein expression in Clone-9 cells (rat hepatocytes). The ataxia telangiectasia mutated (ATM)/rad3-related (ATR)-p53-p21(WAF1) and the phosphatidylinositol-3-kinase (PI3K)-mammalian target of rapamycin (mTOR) pathways are involved in the DNA damage response and the pathogenesis of cancers. Thus, we studied the role of ATM/ATR and PI3K in Arecoline-induced p53 and p21(WAF1) protein expression in Clone-9 cells. We found that Arecoline (0.5 mM) activated the ATM/ATR kinase at 30 min. The Arecoline-activated ATM/ATR substrate contained p-p53Ser15. Moreover, Arecoline only increased the levels of the p-p53Ser6, p-p53Ser15, and p-p53Ser392 phosphorylated p53 isoforms among the known isoforms. ATM shRNA attenuated Arecoline-induced p-p53Ser15 and p21(WAF1) at 24 h. Arecoline (0.5 mM) increased phosphorylation levels of p-AktSer473 and p-mTORSer2448 at 30-60 min. Dominant-negative PI3K plasmids attenuated Arecoline-induced p21(WAF1), but not p-p53Ser15, at 24 h. Rapamycin attenuated Arecoline-induced phosphrylated p-p53Ser15, but not p21(WAF1), at 24 h. ATM shRNA, but not dominant-negative PI3K plasmids, attenuated Arecoline-induced p21(WAF1) gene transcription. We conclude that Arecoline activates the ATM/ATR-p53-p21(WAF1) and the PI3K/Akt-mTOR-p53 pathways in Clone-9 cells. Arecoline-induced phosphorylated p-p53Ser15 expression is dependent on ATM whereas Arecoline-induced p21(WAF1) protein expression is dependent on ATM and PI3K. Moreover, p21(WAF1) gene is transcriptionally induced by Arecoline-activated ATM.
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Arecoline-induced growth arrest and p21WAF1 expression are dependent on p53 in rat hepatocytes.
Toxicology, 2007Co-Authors: Wen-wen Chou, Jinn-yuh Guh, Jung-fa Tsai, Chi-ching Hwang, Hung-chun Chen, Jau-shyang Huang, Yu-lin Yang, Wen-chun Hung, Lea-yea ChuangAbstract:Betel-quid use is associated with the risk of liver cirrhosis and hepatocellular carcinoma and Arecoline, the major alkaloid of betel-quid, is hepatotoxic in mice. Therefore, we studied the cytotoxic and genotoxic effects of Arecoline in normal rat hepatocytes (Clone-9 cells). Arecoline dose-dependently (0.1-1mM) decreased cell cycle-dependent proliferation while inducing DNA damage at 24h. Moreover, Arecoline (1mM)-induced apoptosis and necrosis at 24h. Arecoline dose-dependently (0.1-0.5mM) increased transforming growth factor-beta (TGF-beta) mRNA, gene transcription and bioactivity and neutralizing TGF-beta antibody attenuated Arecoline (0.5mM)-inhibited cell proliferation at 24h. Arecoline (0.5mM) also increased p21(WAF1) protein expression and p21(WAF1) gene transcription. Moreover, Arecoline (0.5mM) time-dependently (8-24h) increased p53 serine 15 phosphorylation. Pifithrin-alpha (p53 inhibitor) and the loss of the two p53-binding elements in the p21(WAF1) gene promoter attenuated Arecoline-induced p21(WAF1) gene transcription at 24h. Pifithrin-alpha also attenuated Arecoline (0.5mM)-inhibited cell proliferation at 24h. We concluded that Arecoline induces cytotoxicity, DNA damage, G(0)/G(1) cell cycle arrest, TGF-beta1, p21(WAF1) and activates p53 in Clone-9 cells. Moreover, Arecoline-induced p21(WAF1) is dependent on p53 while Arecoline-inhibited growth is dependent on both TGF-beta and p53.
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Arecoline-induced growth arrest and p21WAF1 expression are dependent on p53 in rat hepatocytes.
Toxicology, 2007Co-Authors: Wen-wen Chou, Jinn-yuh Guh, Jung-fa Tsai, Chi-ching Hwang, Hung-chun Chen, Jau-shyang Huang, Yu-lin Yang, Wen-chun Hung, Lea-yea ChuangAbstract:Abstract Betel-quid use is associated with the risk of liver cirrhosis and hepatocellular carcinoma and Arecoline, the major alkaloid of betel-quid, is hepatotoxic in mice. Therefore, we studied the cytotoxic and genotoxic effects of Arecoline in normal rat hepatocytes (Clone-9 cells). Arecoline dose-dependently (0.1–1 mM) decreased cell cycle-dependent proliferation while inducing DNA damage at 24 h. Moreover, Arecoline (1 mM)-induced apoptosis and necrosis at 24 h. Arecoline dose-dependently (0.1–0.5 mM) increased transforming growth factor-β (TGF-β) mRNA, gene transcription and bioactivity and neutralizing TGF-β antibody attenuated Arecoline (0.5 mM)-inhibited cell proliferation at 24 h. Arecoline (0.5 mM) also increased p21 WAF1 protein expression and p21 WAF1 gene transcription. Moreover, Arecoline (0.5 mM) time-dependently (8–24 h) increased p53 serine 15 phosphorylation. Pifithrin-α (p53 inhibitor) and the loss of the two p53-binding elements in the p21 WAF1 gene promoter attenuated Arecoline-induced p21 WAF1 gene transcription at 24 h. Pifithrin-α also attenuated Arecoline (0.5 mM)-inhibited cell proliferation at 24 h. We concluded that Arecoline induces cytotoxicity, DNA damage, G 0 /G 1 cell cycle arrest, TGF-β1, p21 WAF1 and activates p53 in Clone-9 cells. Moreover, Arecoline-induced p21 WAF1 is dependent on p53 while Arecoline-inhibited growth is dependent on both TGF-β and p53.
Yu-chao Chang - One of the best experts on this subject based on the ideXlab platform.
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Cytotoxic and non‐genotoxic effects of Arecoline on human buccal fibroblasts in vitro
Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pat, 2007Co-Authors: Yu-chao Chang, Kuo-wei Tai, Min-hsiung Cheng, Lin Shin-shen Chou, Ming-yung ChouAbstract:Chang YC, Tai KW, Cheng MH, Chou LSS, Chou MY: Cytotoxic and non-genotoxic effects of Arecoline on human buccal fibroblasts in vitro. J Oral Pathol Med 1998; 27: 68–71. © Munksgaard, 1998. Betel quid chewing has been linked to oral submucous fibrosis and oral cancer. Cytotoxicity and genotoxicity assays were used to investigate the pathobiologi-cal effects of Arecoline on cultured human buccal fibroblasts. Arecoline increased double-stranded polynucleic acid at the concentration of 0.1 to 10 μg/ml in a concentration-dependent manner. At a concentration higher than 50 μg/ml, Arecoline was cytotoxic to cultured fibroblasts and the cytotoxicity was dose-dependent. No genotoxicity for Arecoline was found even at a concentration of 400 μg/ml. On the other hand, 600 μg/ml glutathione (GSH) and 200 μg/ml glycyrrhizin could prevent the Arecoline-induced cytotoxicity. These results indicate that Arecoline is a cytotoxic agent and no genotoxicity was found to human buccal fibroblasts. Furthermore, increasing consumption of GSH- and glycyrrhizin-rich foods may reduce the oral diseases associated with betel quid chewing.
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cytotoxic and non genotoxic effects of Arecoline on human buccal fibroblasts in vitro
Journal of Oral Pathology & Medicine, 2007Co-Authors: Yu-chao Chang, Kuo-wei Tai, Min-hsiung Cheng, Lin Shin-shen Chou, Ming-yung ChouAbstract:Chang YC, Tai KW, Cheng MH, Chou LSS, Chou MY: Cytotoxic and non-genotoxic effects of Arecoline on human buccal fibroblasts in vitro. J Oral Pathol Med 1998; 27: 68–71. © Munksgaard, 1998. Betel quid chewing has been linked to oral submucous fibrosis and oral cancer. Cytotoxicity and genotoxicity assays were used to investigate the pathobiologi-cal effects of Arecoline on cultured human buccal fibroblasts. Arecoline increased double-stranded polynucleic acid at the concentration of 0.1 to 10 μg/ml in a concentration-dependent manner. At a concentration higher than 50 μg/ml, Arecoline was cytotoxic to cultured fibroblasts and the cytotoxicity was dose-dependent. No genotoxicity for Arecoline was found even at a concentration of 400 μg/ml. On the other hand, 600 μg/ml glutathione (GSH) and 200 μg/ml glycyrrhizin could prevent the Arecoline-induced cytotoxicity. These results indicate that Arecoline is a cytotoxic agent and no genotoxicity was found to human buccal fibroblasts. Furthermore, increasing consumption of GSH- and glycyrrhizin-rich foods may reduce the oral diseases associated with betel quid chewing.
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elevated vimentin expression in buccal mucosal fibroblasts by Arecoline in vitro as a possible pathogenesis for oral submucous fibrosis
Oral Oncology, 2002Co-Authors: Yu-chao Chang, Kuo-wei Tai, Ming-yung Chou, Chung-hung Tsai, S F Yang, Chong-kuei LiiAbstract:Areca quid chewing is strongly correlated with oral submucous fibrosis (OSF) in Taiwan. The cytotoxicity of Arecoline, a major areca nut alkaloid, on human oral fibroblasts has been extensively studied. To date, however, there has been little research exploring the possible effects of Arecoline on cytoskeleton components. In this study, in addition to conducting a cytotoxicity assay, we examine the effect of Arecoline on vimentin, an intermediate filament, and its expression in human buccal mucosal fibroblasts on exposure to various levels of Arecoline (0–200 μg/ml) for 48 h. At a concentration above 50 μg/ml, Arecoline demonstrated dose-dependent cytotoxicity (P<0.05) for cultured fibroblasts. Using sodium dodecyl sulphate-polyacrylamide gel electrophoresis, we demonstrated dose-dependent elevation of 57 kDa cytoskeletal-protein levels for Arecoline. Evidence from immunoblotting assay indicated this 57 kDa cytoskeletal protein was vimentin. The increase in vimentin with Arecoline exposure corresponded to that noted for fibroblasts cultured from OSF patients. Immunohistochemical assay also revealed that vimentin expression was much higher for OSF specimens than for normal buccal mucosa. We suggest these results may advance understanding of the possible pathogenesis for submucous fibrosis through the transformation of normal buccal mucosa as a result of areca quid chewing.
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Adverse effects of Arecoline and nicotine on human periodontal ligament fibroblasts in vitro
Journal of clinical periodontology, 2001Co-Authors: Yu-chao Chang, Chong-kuei Lii, Kuo-wei Tai, Ming-yung ChouAbstract:BACKGROUND, AIMS The habit of betel nut chewing impinges on the daily lives of approximately 200 million people. Betel quid chewers have a higher prevalence of periodontal diseases than non-chewers. This study examined the pathobiological effects of Arecoline, a major component of the betel nut alkaloids, on human periodontal ligament fibroblasts (PDLF) in vitro. METHOD Cell viability, proliferation, protein synthesis, and cellular thiol levels were used to investigate the effects of human PDLF exposed to Arecoline levels of 0 to 200 microg/ml. In addition, nicotine was added to test how it modulated the effects of Arecoline. RESULTS Arecoline significantly inhibited cell proliferation in a dose-dependent manner. At concentrations of 10 and 30 microg/ml, Arecoline suppressed the growth of PDLF by 20% and 50% (p < 0.05), respectively. Arecoline also decreased protein synthesis in a dose-dependent manner during a 24-h culture period. A 100 microg/ ml concentration level of Arecoline significantly inhibited protein synthesis to only 50% of that in the untreated control (p < 0.05). Moreover, Arecoline significantly depleted intracellular thiols in a dose-dependent manner. At concentrations of 25 microg/ml and 100 microg/ml, Arecoline depleted about 18% and 56% of thiols (p < 0.05), respectively. This suggests that Arecoline itself might augment the destruction of periodontium associated with betel nut use. Furthermore, the addition of nicotine acted with a synergistic effect on the Arecoline-induced cytotoxicity. At a concentration of 60 microg/ml, Arecoline suppressed the growth of PDLF by about 33% and 5 mM nicotine enhanced the Arecoline-induced cytotoxic response to cause about 66% cell death. CONCLUSION During thiol depletion, Arecoline may render human PDLF more vulnerable to reactive agents within cigarettes. Taken together, people who combine habits of betel nut chewing with cigarette smoking could be more susceptible to periodontium damage than betel nut chewing alone.
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synergistic effects of nicotine on Arecoline induced cytotoxicity in human buccal mucosal fibroblasts
Journal of Oral Pathology & Medicine, 2001Co-Authors: Yu-chao Chang, Chong-kuei Lii, Kuo-wei Tai, Tsuihwa Tseng, Ming-yung ChouAbstract:Areca quid chewing has been linked to oral submucous fibrosis and oral cancer. Arecoline, a major areca nut alkaloid, is considered to be the most important etiologic factor in the areca nut. In order to elucidate the pathobiological effects of Arecoline, cytotoxicity assays, cellular glutathione S-transferase (GST) activity and lipid peroxidation assay were employed to investigate cultured human buccal mucosal fibroblasts. To date, there is a large proportion of areca quid chewers who are also smokers. Furthermore, nicotine, the major product of cigarette smoking, was added to test how it modulated the cytotoxicity of Arecoline. At a concentration higher than 50 microg/ml, Arecoline was shown to be cytotoxic to human buccal fibroblasts in a dose-dependent manner by the alamar blue dye colorimetric assay (P<0.05). In addition, Arecoline significantly decreased GST activity in a dose-dependent manner (P<0.05). At concentrations of 100 microg/ml and 400 microg/ml, Arecoline reduced GST activity about 21% and 46%, respectively, during a 24 h incubation period. However, Arecoline at any test dose did not increase lipid peroxidation in the present human buccal fibroblast test system. The addition of extracellular nicotine acted synergistically on the Arecoline-induced cytotoxicity. Arecoline at a concentration of 50 microg/ml caused about 30% of cell death over the 24 h incubation period. However, 2.5 mM nicotine enhanced the cytotoxic response and caused about 50% of cell death on 50 microg/ml Arecoline-induced cytotoxicity. Taken together, Arecoline may render human buccal mucosal fibroblasts more vulnerable to other reactive agents in cigarettes via GST reduction. The compounds of tobacco products may act synergistically in the pathogenesis of oral mucosal lesions in areca quid chewers. The data presented here may partly explain why patients who combined the habits of areca quid chewing and cigarette smoking are at greater risk of contracting oral cancer.
Yung-hsi Kao - One of the best experts on this subject based on the ideXlab platform.
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Arecoline inhibits the growth of 3t3 l1 preadipocytes via amp activated protein kinase and reactive oxygen species pathways
PLOS ONE, 2018Co-Authors: Zi-han Tian, Jueng-tsueng Weng, Li-jane Shih, An-ci Siao, Tsai-yun Chan, Yi-wei Tsuei, Yow-chii Kuo, Tsu-shing Wang, Yung-hsi KaoAbstract:The present study was designed to investigate the pathways involved in the effect of betel nut Arecoline on cell viability in 3T3-L1 preadipocytes. Arecoline, but not arecaidine or guvacine, inhibited preadipocyte viability in a concentration- and time-dependent manner. Arecoline arrested preadipocyte growth in the G2/M phase of the cell cycle; decreased the total levels of cyclin-dependent kinase 1 (CDK1), p21, and p27 proteins; increased p53 and cyclin B1 protein levels; and had no effect on CDK2 protein levels. These results suggested that Arecoline selectively affected a particular CDK subfamily. Arecoline inhibited AMP-activated protein kinase (AMPK) activity; conversely, the AMPK activator, AICAR, blocked the Arecoline-induced inhibition of cell viability. Pre-treatment with the antioxidant, N-acetylcysteine, prevented the actions of Arecoline on cell viability, G2/M growth arrest, reactive oxygen species (ROS) production, and the levels of CDK1, p21, p27, p53, cyclin B1, and phospho-AMPK proteins. These AMPK- and ROS-dependent effects of Arecoline on preadipocyte growth may be related to the mechanism underlying the modulatory effect of Arecoline on body weight.
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Betel nut alkaloids reduced cell viability of primary preadipocytes isolated from the epididymal adipose tissues of male mice after 48 h of treatment, and the effect of Arecoline depended on AMPK and ROS pathways.
2018Co-Authors: Zi-han Tian, Jueng-tsueng Weng, Li-jane Shih, An-ci Siao, Tsai-yun Chan, Yi-wei Tsuei, Yow-chii Kuo, Tsu-shing Wang, Yung-hsi KaoAbstract:Data are expressed as the mean ± SEM from triplicate determinations. +, p < 0.1 vs. control; *, p < 0.05 vs. control; #. p < 0.05, Arecoline vs. arecaidine, Arecoline vs. guvacine, Arecoline vs. NAC + Arecoline, or Arecoline vs. AICAR + Arecoline (bracket); §, p < 0.05, 100 μM vs. 400 μM.
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The effect of Arecoline on cell viability in 3T3-L1 preadipocyte was dependent on the AMP-activated protein kinase (AMPK) pathway.
2018Co-Authors: Zi-han Tian, Jueng-tsueng Weng, Li-jane Shih, An-ci Siao, Tsai-yun Chan, Yi-wei Tsuei, Yow-chii Kuo, Tsu-shing Wang, Yung-hsi KaoAbstract:(A) Arecoline dose-dependently reduced protein amounts of phospho-AMPK (pAMPK) but not the total amount of AMPK protein. (B) An alkaloid-specific effect of betel nut on AMPK phosphorylation was observed after 24 h of 400-μM treatment. (C) The AMPK activator AICAR at doses of 125 and 250 μM antagonized the Arecoline-reduced cell viability of preadipocytes, while its inhibitor compound C (10 μM) alone inhibited cell viability but did not block Arecoline-reduced cell viability. Total AMPK was measured by Western blot analysis and then expressed after normalization to actin, whereas pAMPK was normalized to its total AMPK. Data are expressed as the mean ± SEM from triplicate determinations. *, p < 0.05 vs. the control; #, p < 0.05 Arecoline vs. Arecoline + AICAR.
Hans Zoellner - One of the best experts on this subject based on the ideXlab platform.
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Arecoline increases basic fibroblast growth factor but reduces expression of il 1 il 6 g csf and gm csf in human umbilical vein endothelium
Journal of Oral Pathology & Medicine, 2015Co-Authors: Mafaz Ullah, Stephen Cox, Hans Zoellner, Elizabeth Kelly, Malcolm A S MooreAbstract: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.
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Arecoline is cytotoxic for human endothelial cells
Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pat, 2014Co-Authors: Mafaz Ullah, Stephen Cox, Elizabeth Kelly, Ross A. Boadle, Hans ZoellnerAbstract:Background Oral submucous fibrosis is a pre-malignant fibrotic condition caused by areca nut use and involves reduced mucosal vascularity. Arecoline is the principal areca nut alkaloid and is cytotoxic for epithelium and fibroblasts. Endothelial cell cycle arrest is reported on exposure to Arecoline, as is cytotoxicity for endothelial–lung carcinoma hybrid cells. We here describe cytotoxicity for primary human endothelial cultures from seven separate donors. Materials and methods Human umbilical vein endothelial cells were exposed to increasing concentrations of Arecoline and examined by: phase-contrast microscopy, haemocytometer counts, transmission electron microscopy, lactate dehydrogenase release and the methyl-thiazol-tetrazolium assay. Results Vacuolation and detachment of endothelium were observed at and above Arecoline concentrations of 333 μg/ml or more. Ultrastructural features of cellular stress were seen after 24-h treatment with 111 μg/ml Arecoline and included reduced ribosomal studding of endoplasmic reticulum, increased autophagolysosomal structures, increased vacuolation and reduced mitochondrial cristae with slight swelling. Similar changes were seen at 4 h with Arecoline at 333 μg/ml or above, but with more severe mitochondrial changes including increased electron density of mitochondrial matrix and greater cristal swelling, while by 24 h, these cells were frankly necrotic. Haemocytometer counts were paralleled by both lactate dehydrogenase release and the methyl-thiazol-tetrazolium assays. Conclusions Arecoline is cytotoxic via necrosis for endothelium, while biochemical assays indicate no appreciable cellular leakage before death and detachment, as well as no clear effect on mitochondrial function in viable cells. Arecoline toxicity may thus contribute to reduced vascularity in oral submucous fibrosis.
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salivary Arecoline levels during areca nut chewing in human volunteers
Journal of Oral Pathology & Medicine, 2010Co-Authors: Stephen Cox, Edward R Vickers, Sonia Ghu, Hans ZoellnerAbstract:J Oral Pathol Med (2010) 39: 465–469 Background: Arecoline stimulates cultured cells above 0.1 μg/ml and is cytotoxic above 10 μg/ml. Although this alkaloid seems important for areca nut induced oral carcinogenesis, little is known of the levels achieved during chewing. Materials and methods: Saliva was collected in 3- to 5-min intervals over 50 min in 32 habitual chewers: before, for 25 min during, and for 20 min after chewing areca nut (0.5 g) without any other additives. Salivary Arecoline was quantitated by HPLC-MS. Controls comprised six subjects who denied areca nut use, and who were given rubber-base material to chew during experiments instead. Results: Arecoline was detected before chewing in 22 subjects, exceeding the 0.1 μg/ml threshold in 20 cases. Salivary Arecoline exceeded either the 0.1 or 10 μg/ml thresholds in all participants during chewing (P < 0.001). Maximum concentrations ranged from 5.66 to 97.39 μg/ml. All subjects reached 0.1 μg/ml salivary Arecoline in at least 85% of time points studied (P < 0.0001), whereas 10 μg/ml was reached in 11 participants in at least 30% of the time points (P < 0.003). Arecoline concentrations varied greatly over time between individuals, and levels were much lower when peak concentrations were reached before 3 min, than in cases where Arecoline peaked later (P < 0.02). No salivary Arecoline was found in control saliva. Conclusions: Areca nut users have persistent background salivary Arecoline levels long after chewing, whereas concentrations achieved are highly variable and consistent with a role in oral pre-malignancy and malignancy.
