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A comprehensive analysis of metabolomics and lipidomics in areca nut mediated oral submucous fibrosis progression through LCMS and Raman spectroscopy

ABSTRACT: The current study aims to evaluate the biomolecular alterations in fibroblast cells after treatment with areca nut. An aqueous extract of areca nut was prepared and characterized. To check whether the cells treated with areca nut mimic the OSMF, cell viability, proliferation, migration, and morphologic changes were observed in these cells up to 48 Hrs after treatment. We used Raman microspectroscopy and LC-MS techniques to evaluate the biomolecular changes in ANE-treated cells. The possible interactions and biomolecular pathways associated with these molecules were also explored.

ORGANISM(S): Homo sapiens (human)

SUBMITTER:

PROVIDER: S-BSST1088 | biostudies-other |

REPOSITORIES: biostudies-other

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Ultrasonographic evaluation of oral submucous fibrosis in habitual areca nut chewers.

Project description:OBJECTIVES: To characterize the sonographic features of the buccal mucosa in patients with oral submucous fibrosis (OSF). METHODS: Three groups (controls with areca-related habits, controls without areca-related habits and clinically diagnosed OSF cases), each comprising 30 subjects, were included in the study. After a thorough clinical examination, transcutaneous B-mode ultrasonography was performed with a multifrequency linear transducer (5-10 MHz) for anterior and posterior buccal mucosa bilaterally. Both clinical and ultrasound findings were recorded by three independent observers. One-way analysis of variance and Tukey's honestly significant difference post-hoc tests were used for statistical comparisons between groups and Pearson χ(2) tests to compare the proportions. Kappa statistics was used to determine the interobserver agreement. RESULTS: The submucosa that appeared hypoechoic in the control groups had significantly increased echogenicity in the case group (hypo- to isoechoic in 46.7% and isoechoic in 53.3%). The differentiation between the submucosa and the muscle layer appeared distinct in the control groups while it was not clear in the case group (indistinct in 50% and completely lost in 50%). The number of sites found positive on the ultrasound was significantly greater than the number of clinically positive sites. There was a very good interobserver consistency in clinical and ultrasound findings. CONCLUSIONS: Ultrasonography of the buccal mucosa demonstrates increased submucosal echogenicity and reduced echo differentiation between submucosa and muscle layer in OSF cases. Hence, it can be used as a non-invasive imaging modality to assess the disease extent and severity across the entire buccal mucosa to supplement clinical evaluation.

| S-EPMC3828018 | biostudies-other

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

Project description: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 IC(50) 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.

| S-EPMC4594681 | biostudies-literature

Activation of TGF-β pathway by areca nut constituents: a possible cause of oral submucous fibrosis.

Project description:Oral submucous fibrosis (OSF) is a chronic inflammatory disease characterized by the accumulation of excess collagen, and areca nut chewing has been proposed as an important etiological factor for disease manifestation. Activation of transforming growth factor-β signaling has been postulated as the main causative event for increased collagen production in OSF. Oral epithelium plays important roles in OSF, and arecoline has been shown to induce TGF-β in epithelial cells. In an attempt to understand the role of areca nut constituents in the manifestation of OSF, we studied the global gene expression profile in epithelial cells (HaCaT) following treatment with areca nut water extract or TGF-β. Interestingly, 64% of the differentially regulated genes by areca nut water extract matches with the TGF-β induced gene expression profile. Out of these, expression of 57% of genes was compromised in the presence of ALK5 (TβRI) inhibitor and 7% were independently induced by areca nut, highlighting the importance of TGF-β in areca nut actions. Areca nut water extract treatment induced p-SMAD2 and TGF-β downstream targets in HaCaT cells but not in human gingival fibroblast cells (hGF), suggesting epithelial cells could be the source of TGF-β in promoting OSF. Water extract of areca nut consists of polyphenols and alkaloids. Both polyphenol and alkaloid fractions of areca nut were able to induce TGF-β signaling and its downstream targets. Also, SMAD-2 was phosphorylated following treatment of HaCaT cells by Catechin, Tannin and alkaloids namely Arecoline, Arecaidine and Guvacine. Moreover, both polyphenols and alkaloids induced TGF-β2 and THBS1 (activator of latent TGF-β) in HaCaT cells suggesting areca nut mediated activation of p-SMAD2 involves up-regulation and activation of TGF-β. These data suggest a major causative role for TGF-β that is induced by areca nut in OSF progression.

| S-EPMC3526649 | biostudies-literature

Role of Areca Nut Induced TGF-β and Epithelial-Mesenchymal Interaction in the Pathogenesis of Oral Submucous Fibrosis.

Project description:Areca nut consumption has been implicated in the progression of Oral Submucous fibrosis (OSF); an inflammatory precancerous fibrotic condition. Our previous studies have demonstrated the activation of TGF-β signaling in epithelial cells by areca nut components and also propose a role for epithelial expressed TGF-β in the pathogenesis of OSF. Although the importance of epithelial cells in the manifestation of OSF has been proposed, the actual effectors are fibroblast cells. However, the role of areca nut and TGF-β in the context of fibroblast response has not been elucidated. Therefore, to understand their role in the context of fibroblast response in OSF pathogenesis, human gingival fibroblasts (hGF) were treated with areca nut and/or TGF-β followed by transcriptome profiling. The gene expression profile obtained was compared with the previously published transcriptome profiles of OSF tissues and areca nut treated epithelial cells. The analysis revealed regulation of 4666 and 1214 genes by areca nut and TGF-β treatment respectively. The expression of 413 genes in hGF cells was potentiated by areca nut and TGF-β together. Further, the differentially expressed genes of OSF tissues compared to normal tissues overlapped significantly with areca nut and TGF-β induced genes in epithelial and hGF cells. Several positively enriched pathways were found to be common between OSF tissues and areca nut +TGF-β treated hGF cells. In concordance, areca nut along with TGF-β enhanced fibroblast activation as demonstrated by potentiation of αSMA, γSMA and collagen gel contraction by hGF cells. Furthermore, TGF-β secreted by areca nut treated epithelial cells influenced fibroblast activation and other genes implicated in fibrosis. These data establish a role for areca nut influenced epithelial cells in OSF progression by activation of fibroblasts and emphasizes the importance of epithelial-mesenchymal interaction in OSF.

| S-EPMC4479469 | biostudies-literature

Activation of TGF-beta pathway by areca nut constituents: A possible cause of Oral submucous fibrosis.

Project description:To understand the role of areca nut constituents in manifestation of Oral submucous fibrosis, we studied gene expression profile in epithelial cells following areca nut water extract treatment. A comaprison with TGF-beta induced gene expression changes were performed as epithelial cells were predicted to be source of TGF-beta. Control Vs Areca nut 5 ug/ml water extract (5H) (2), Contro Vs TGF-beta (2), Control Vs ALK5 (TbetaRI inhibitor) (2), Control Vs 5H + ALK5 inhibitor (2). (2)- Biological duplicates.

2013-05-22 | E-GEOD-38227 | biostudies-arrayexpress

Activation of TGF-beta pathway by areca nut constituents: A possible cause of Oral submucous fibrosis.

2013-05-22 | GSE38227 | GEO

Role of areca nut induced TGF-β and epithelial mesenchymal interaction in pathogenesis of precancerous Oral submucous fibrosis

Project description:To understand the role of areca nut and TGF-β induced gene expression changes in fibroblasts and its contibution in the manifestation of Oral submucous fibrosis, we studied gene expression profile in primary human gingival fibroblast (hGF) cells following treatment with areca nut, TGF-β and both together. Control Vs Areca nut 5 µg/ml water extract (5H) (2), Contro Vs TGF-β (2), Control Vs Areca nut (5 µg/ml) and TGF-β (5 ng/ml) (5H+T) (2). (2)- Biological duplicates.

2015-07-06 | E-GEOD-59414 | biostudies-arrayexpress

Role of areca nut induced JNK/ATF2/Jun axis in the activation of TGF-β pathway in precancerous Oral Submucous Fibrosis.

Project description:Oral submucous fibrosis (OSF) is potentially premalignant with progressive and irreversible extracellular matrix deposition accompanied by epithelial atrophy and like other fibrotic disorders, is primarily a TGF-β driven disease. OSF is caused by prolonged chewing of areca nut. Our previous studies reported a pivotal role for TGF-β activation and its effects contributing to OSF. However, the mechanism for activation of TGF-β signaling in OSF is still unknown. In this study we demonstrate activation of TGF-β signaling with sub-cytotoxic dose of areca nut in epithelial cells and discovered a key role for pJNK in this process. In good correlation; pJNK was detected in OSF tissues but not in normal tissues. Moreover, activation of JNK was found to be dependent on muscarinic acid receptor induced Ca2+/CAMKII as well as ROS. JNK dependent phosphorylation of ATF2/c-Jun transcription factors resulted in TGF-β transcription and its signaling. pATF2/p-c-Jun were enriched on TGF-β promoter and co-localized in nuclei of epithelial cells upon areca nut treatment. In corroboration, OSF tissue sections also had nuclear pATF2 and p-c-Jun. Our results provide comprehensive mechanistic details of TGF-β signaling induced by etiological agent areca nut in the manifestation of fibrosis which can lead to new therapeutic modalities for OSF.

| S-EPMC5052620 | biostudies-literature

Role of areca nut induced TGF-β and epithelial mesenchymal interaction in pathogenesis of precancerous Oral submucous fibrosis

2015-07-06 | GSE59414 | GEO

Circulating miRNAs in the regulation of epithelial mesenchymal transition and their potential role as possible diagnostic biomarkers in areca nut associated oral submucous fibrosis in South Indian population

Project description:Expression of miRNA analysis in human lecukocytes was analysed using Next-generation sequencing (NGS) method in Oral Submucosis Firbrosis condition.

2018-12-19 | GSE124060 | GEO

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