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Cell Cycle (Georgetown, Tex.) May 2022Arecoline, the most abundant alkaloid of the areca nut, induces toxicity to neurons. Hydrogen sulfide (HS) is an endogenous gas with neuroprotective effects. We recently...
Arecoline, the most abundant alkaloid of the areca nut, induces toxicity to neurons. Hydrogen sulfide (HS) is an endogenous gas with neuroprotective effects. We recently found that arecoline reduced endogenous HS content in PC12 cells. In addition, exogenously administration of HS alleviated the neurotoxicity of arecoline on PC12 cells. Increasing evidence has demonstrated the neuroprotective role of improvement of autophagic flux. Therefore, the aim of the present work is to explore whether improvement of autophagic flux mediates the protection of HS against arecoline-caused neurotoxicity. Transmission electron microscope (TEM) for observation of ultrastructural morphology. Western blotting was used to detect protein expression of the related markers. Functional analysis contained LDH release assay, Hoechst 33,258 nuclear staining and flow cytometry were used to detect cytotoxicity and apoptosis. In the present work, we found that arecoline disrupted autophagy flux in PC12 cells as evidenced by accumulation of autophagic vacuoles, increase in LC3II/LC3I, and upregulation of p62 expression in PC12 cells. Notably, we found that sodium hydrosulfide (NaHS), the donor of HS improved arecoline-blocked autophagy flux in PC12 cells. Furthermore, we found that blocking autophagic flux by chloroquine (CQ), the inhibitor of autophagy flux, antagonized the inhibitory role of NaHS in arecoline-induced cytotoxicity apoptosis and endoplasmic reticulum (ER) stress. In conclusion, HS improves arecoline-caused disruption of autophagic flux to exert its protection against the neurotoxicity of arecoline.
Topics: Animals; Apoptosis; Arecoline; Autophagy; Endoplasmic Reticulum Stress; Hydrogen Sulfide; PC12 Cells; Rats
PubMed: 35316162
DOI: 10.1080/15384101.2022.2040932 -
Journal of Clinical Laboratory Analysis Apr 2022Hepatocellular carcinoma (HCC) is the most common histological subtype of liver cancer and the third leading cause of death from cancer globally. Recent studies...
BACKGROUND
Hepatocellular carcinoma (HCC) is the most common histological subtype of liver cancer and the third leading cause of death from cancer globally. Recent studies suggested cell death is also a key regulator of tumour progression. The purpose of this study was to generate a new predictive signature for HCC patients based on a complete analysis of necroptosis-associated genes.
METHODS
We extracted the mRNA expression profiles of HCC patients from the TCGA and ICGC databases and their clinical data. In addition, we used the IMvigor210 cohort to validate our model molecule's ability to predict the effect of immunotherapy. In the TCGA cohort, a seven-gene risk-prognostic model was constructed using univariate cox-Lasoo regression. External validation was conducted using the ICGC cohort. The ssGSEA algorithm is used to determine the degree of immune function response. The CMAP databases are used for chemotherapy drug analysis and screening for drugs that reduce the expression of high-risk genes. The cbioportal database was used to explore mutations in model genes.
RESULTS
Survival analysis shows shorter survival for high-risk patients. Immune function analysis revealed significant differences in the activity of immune pathways between risk subgroups. Varied risk scores result in dramatically diverse immune infiltration and tumour growth, as well as significantly different chemotherapeutic sensitivity. In addition, Apigenin and LY-294002 reduced the expression of high-risk genes, while Arecoline had the opposite effect. In the immunotherapy IMvigor210 cohort, risk scores were significantly different between the objective responder and non-responder groups. By comparing the models constructed with published literature, it is suggested that our model has better predictive power.
CONCLUSIONS
We created a new prognostic signature of necroptosis-related genes that can be used as potential prognostic biomarkers to guide effective personalized therapy for hepatocellular carcinoma patients.
Topics: Biomarkers, Tumor; Carcinoma, Hepatocellular; Gene Expression Profiling; Humans; Liver Neoplasms; Necroptosis; Prognosis
PubMed: 35293027
DOI: 10.1002/jcla.24346 -
Cancer Science Sep 2022The high prevalence of oral squamous cell carcinoma (OSCC) in South Asia is associated with habitual areca nut chewing. Arecoline, a primary active carcinogen within...
The high prevalence of oral squamous cell carcinoma (OSCC) in South Asia is associated with habitual areca nut chewing. Arecoline, a primary active carcinogen within areca nut extract, is known to promote OSCC pathological development. Dysregulation of N6-methyladenosine (m6A) modification has begun to emerge as a significant contributor to cancer development and progression. However, the biological effects and molecular mechanisms of m6A modification in arecoline-promoted OSCC malignance remain elusive. We reveal that chronic arecoline exposure substantially induces upregulation of fat mass and obesity-associated protein (FTO), MYC, and programmed cell death-ligand 1 (PD-L1) in OSCC cells. Moreover, upregulation of PD-L1 is observed in OSCC cell lines and tissues and is associated with areca nut chewing in OSCC patients. We also demonstrate that arecoline-induced FTO promotes the stability and expression levels of PD-L1 transcripts through mediating m6A modification and MYC activity, respectively. PD-L1 upregulation confers superior cell proliferation, migration, and resistance to T-cell killing to OSCC cells. Blockage of PD-L1 by administration of anti-PD-L1 antibody shrinks tumor size and improves mouse survival by elevating T-cell-mediated tumor cell killing. Therefore, targeting PD-L1 might be a potential therapeutic strategy for treating PD-L1-positive OSCC patients, especially those with habitual areca nut chewing.
Topics: Animals; Apoptosis; Areca; Arecoline; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Immunity; Ligands; Mice; Mouth Neoplasms; Obesity; Squamous Cell Carcinoma of Head and Neck
PubMed: 35289035
DOI: 10.1111/cas.15332 -
Journal of Education and Health... 2022The study aims to measure and compare pulmonary function tests (PFTs) in oral submucous fibrosis (OSMF) patients (smokers/nonsmokers) and normal individuals.
INTRODUCTION
The study aims to measure and compare pulmonary function tests (PFTs) in oral submucous fibrosis (OSMF) patients (smokers/nonsmokers) and normal individuals.
MATERIALS AND METHODS
The study population included 150 participants that comprised 50 nonsmoker OSMF patients, 50 OSMF patients who smoke as well, and 50 patients with no deleterious habits. Spirometer was used to assess PFT.
RESULTS
Results showed that a significant value was obtained for forced vital capacity (FVC), forced expiratory volume in 1 s (FEV1), FEV1/FVC, peak expiratory flow rate (PEFR), and maximum voluntary ventilation (MVV) and also for the predicted values of FEV, FEV1, FEV1/FVC, PEFR, and MVV in OSMF (smokers/nonsmokers) study groups.
CONCLUSION
Thus, the decrease in pulmonary function can be an alarming sign for restrictive type of pulmonary disease.
PubMed: 35281387
DOI: 10.4103/jehp.jehp_587_21 -
International Journal of Molecular... Jan 2022Betel quid (BQ) is a package of mixed constituents that is chewed by more than 600 million people worldwide, particularly in Asia. The formulation of BQ depends on a... (Review)
Review
Betel quid (BQ) is a package of mixed constituents that is chewed by more than 600 million people worldwide, particularly in Asia. The formulation of BQ depends on a variety of factors but typically includes areca nut, betel leaf, and slaked lime and may or may not contain tobacco. BQ chewing is strongly associated with the development of potentially malignant and malignant diseases of the mouth such as oral submucous fibrosis (OSMF) and oral squamous cell carcinoma (OSCC), respectively. We have shown recently that the constituents of BQ vary geographically and that the capacity to induce disease reflects the distinct chemical composition of the BQ. In this review, we examined the diverse chemical constituents of BQ and their putative role in oral carcinogenesis. Four major areca alkaloids-arecoline, arecaidine, guvacoline and guvacine-together with the polyphenols, were identified as being potentially involved in oral carcinogenesis. Further, we propose that fibroblast senescence, which is induced by certain BQ components, may be a key driver of tumour progression in OSMF and OSCC. Our study emphasizes that the characterization of the detrimental or protective effects of specific BQ ingredients may facilitate the development of targeted BQ formulations to prevent and/or treat potentially malignant oral disorders and oral cancer in BQ users.
Topics: Areca; Arecoline; Carcinoma, Squamous Cell; Disease Progression; Humans; Mouth Neoplasms; Nicotinic Acids; Oral Submucous Fibrosis; Plant Extracts
PubMed: 35163557
DOI: 10.3390/ijms23031637 -
Contemporary Clinical Dentistry 2021Oral submucous fibrosis (OSF) is extensively prevalent in India and South-East Asia owing to the habit of arecanut (AN) use. Epithelial atrophy, hyposalivation, and...
BACKGROUND AND OBJECTIVES
Oral submucous fibrosis (OSF) is extensively prevalent in India and South-East Asia owing to the habit of arecanut (AN) use. Epithelial atrophy, hyposalivation, and immune alterations in OSF may predispose to increased Candidal carriage. Stomatopyrosis in OSF can result from multiple causes, which may also include Candidal infection. Hence, this study is aimed to assess Candidal carriage, species characterization, salivary flow rate (SFR) and its relationship with the clinical features (stomatopyrosis and mouth opening [MO]) in OSF patients alongwith the response to antifungal treatment in patients with higher Candidal carriage.
METHODOLOGY
In this case-control study, 60 OSF patients and 30 age- and sex-matched control subjects were enrolled. SFR was assessed using modified Schirmer test. Samples for Candidal assessment were collected with the oral rinse technique and cultured. The isolated yeast species were counted and identified based on Gram staining, germ tube test, and CHROMagar. Data were analyzed with Chi-square test, Pearson's correlation test, and one-way ANOVA test.
RESULTS
The distribution of mean visual analog scale (VAS) score, SFR and MO was significantly varied ( < 0.001) in the study and control groups. was found to be present significantly ( = 0.048) in OSF group as compared to control group. was the predominant species. No statistically significant association was obtained regarding Candidal isolation and SFR, burning sensation and MO in OSF patients. Only 1 patient in the study group yielded a high Candidal carriage (>400 CFU/mL) and reported relief in burning sensation (VAS score) with antifungal therapy.
CONCLUSIONS
OSF patients yielded a significant higher oral Candidal carriage. Although it was not found to be associated directly, its role as a "cause and effect" in SFR and clinical features (stomatopyrosis and MO) of OSF cannot be ignored.
PubMed: 35068834
DOI: 10.4103/ccd.ccd_296_20 -
Journal of Pharmacy & Bioallied Sciences Nov 2021The present study aimed to establish cell lines of fibroblast from human OSF tissues and their response to varying concentrations of arecoline. The various morphological...
OBJECTIVE
The present study aimed to establish cell lines of fibroblast from human OSF tissues and their response to varying concentrations of arecoline. The various morphological forms of fibroblasts were identified to establish phenotypic change.
MATERIALS AND METHOD
Fibroblast cell lines were obtained from control samples as well as from OSF cases. The cell lines were treated with 50/100/150/300/500 ug/ml of arecoline and morphology were determined.
RESULTS
Three morphological forms were detected; F1 spindle, F2 epitheloid and the F3 stellate. The F3 to F1 ratio was higher in OSF. Arecoline at 50ug/ml was stimulatory and at 150ug/ml cytotoxic to the cell lines.
CONCLUSION
Arecoline seems to enhance proliferation of the fibroblast at lower concentrations but cytotoxic at higher levels. This is probably due to the generation of new cell lines and response of the arecoline receptors indicating phenotypic change.
PubMed: 35017962
DOI: 10.4103/jpbs.jpbs_408_21 -
Drug Design, Development and Therapy 2021Betel nuts have long been used in traditional Chinese medicine. In our study, the bioactive components of betel nut were systematically investigated, and the main...
BACKGROUND
Betel nuts have long been used in traditional Chinese medicine. In our study, the bioactive components of betel nut were systematically investigated, and the main components and their target genes in the treatment of depression were predicted.
METHODS
The metabolites of the kernels and peels were analyzed with a UPLC-MS/MS system. Mass spectrometry outcomes were annotated by MULTIAQUANT. "Compound-disease targets" were utilized to construct a pharmacology network.
RESULTS
A total of 873 metabolites were identified, with a high abundance of flavonoids, alkaloids, and phenols. Moreover, the abundance of flavonoids, alkaloids, and phenols in the kernel was significantly higher than that in the peel. A high abundance of catechin, arginine, and phenylalanine was detected in the kernel, while a high abundance of arginine, arecoline, and aminobutyric acid was detected in the peel. Catechins and cyanoside were the most abundant flavonoids in the kernel and peel, respectively. Arecoline was the most abundant alkaloid. A total of 111 metabolites showed a significant difference between the kernels and peels. The relative abundance of 40 differential metabolites was higher than 100,000, including 14 primary metabolites, 12 flavonoids, 4 phenols, and 4 alkaloids. Among the 40 high abundance metabolites, 20 were higher in the kernel and 20 in the peel. In addition, the enrichment of metabolic pathways found that the kernel and peel of the fruit adopted different metabolic pathways for the synthesis of flavonoids and alkaloids. Network pharmacology prediction showed that 93 metabolites could target 141 depression-related genes. The main components of betel nut intervention in depression were predicted to include L-phenylalanine, protocatechuic acid, okanin, nicotinic acid, L-tyrosine, benzocaine, syringic acid, benzocaine, phloretic acid, cynaroside, and 3,4-dihydroxybenzaldehyde.
CONCLUSION
Betel nuts are rich in natural metabolites, and some of these metabolites can participate in the intervention of depression. In addition, the metabolites showed distinct characteristics between the kernel and peel. Therefore, it is necessary to comprehensively and rationally use betel nuts.
Topics: Alkaloids; Antidepressive Agents; Areca; Chromatography, High Pressure Liquid; Computational Biology; Depression; Flavonoids; Humans; Metabolomics; Network Pharmacology; Phenols; Tandem Mass Spectrometry
PubMed: 34880597
DOI: 10.2147/DDDT.S335312 -
ACS Omega Nov 2021Tobacco use is the leading preventable cause of premature disease and death in the United States. Approximately, 34 million U.S. adults currently smoke cigarettes. We...
Automated Solid Phase Extraction and Polarity-Switching Tandem Mass Spectrometry Technique for High Throughput Analysis of Urine Biomarkers for 14 Tobacco-related Compounds.
Tobacco use is the leading preventable cause of premature disease and death in the United States. Approximately, 34 million U.S. adults currently smoke cigarettes. We developed a method for automated sample preparation and liquid chromatography-tandem mass spectrometry quantitation of 14 tobacco-related analytes: nicotine (NICF), cotinine (COTF), -3'-hydroxycotinine (HCTF), menthol glucuronide (MEG), anabasine (ANBF), anatabine (ANTF), isonicoteine (ISNT), myosmine (MYOS), beta-nicotyrine (BNTR), bupropion (BUPR), cytisine (CYTI), varenicline (VARE), arecaidine (ARD), and arecoline (ARL). The method includes automated solid-phase extraction using customized positive-pressure functions. The preparation scheme has the capacity to process a batch of 96 samples within 4 h with greater than 88% recovery for all analytes. The 14 analytes, separated within 4.15 min using reversed-phase liquid chromatography, were determined using a triple-quadrupole mass spectrometer with atmospheric-pressure chemical ionization and multiple reaction monitoring in negative and positive ionization modes. Wide quantitation ranges, within 1.2-72,000 ng/mL, were established especially for COTF, HCTF, MEG, and NICF to quantify the broad range of biomarker concentrations found in the U.S. population. The method accuracy is above 90% while the overall imprecision is below 7%. Finally, we tested urine samples from 90 smokers and observed detection rates of over 98% for six analytes with urinary HCTF and MEG concentrations ranging from 200-14,100 and 60-57,100 ng/mL, respectively. This high throughput analytical process can prepare and analyze a sample in 9 min and along with the 14-compound analyte panel can be useful for tobacco-exposure studies, in smoking-cessation programs, and for detecting changes in exposure related to tobacco products and their use.
PubMed: 34841133
DOI: 10.1021/acsomega.1c02543 -
Frontiers in Pharmacology 2021Chewing areca nut (betel quid) is strongly associated with oral submucous fibrosis (OSF), a pre-cancerous lesion. Among the areca alkaloids, arecoline is the main agent...
Chewing areca nut (betel quid) is strongly associated with oral submucous fibrosis (OSF), a pre-cancerous lesion. Among the areca alkaloids, arecoline is the main agent responsible for fibroblast proliferation; however, the specific molecular mechanism of arecoline affecting the OSF remains unclear. The present study revealed that arecoline treatment significantly enhanced Transforming growth factor-β (TGF-β)-induced buccal mucosal fibroblast (BMF) activation and fibrotic changes. Arecoline interacts with phosphodiesterase 4A (PDE4A) to exert its effects through modulating PDE4A activity but not PDE4A expression. PDE4A silence reversed the effects of arecoline on TGF-β-induced BMFs activation and fibrotic changes. Moreover, the exchange protein directly activated by cAMP 1 (Epac1)-selective Cyclic adenosine 3',5'-monophosphate (cAMP) analog (8-Me-cAMP) but not the protein kinase A (PKA)-selective cAMP analog (N6-cAMP) remarkably suppressed α-smooth muscle actin(α-SMA) and Collagen Type I Alpha 1 Chain (Col1A1) protein levels in response to TGF-β1 and arecoline co-treatment, indicating that cAMP-Epac1 but not cAMP-PKA signaling is involved in arecoline functions on TGF-β1-induced BMFs activation. In conclusion, arecoline promotes TGF-β1-induced BMFs activation through enhancing PDE4A activity and the cAMP-Epac1 signaling pathway during OSF. This novel mechanism might provide more powerful strategies for OSF treatment, requiring further and clinical investigation.
PubMed: 34819854
DOI: 10.3389/fphar.2021.722040